Bax protein as a prognostic marker in colorectal cancer: characteristics, role in apoptosis regulation, and correlation with disease prognosis (literature review)

Background: Generally, Semaphorins are secretary or transmembrane-bound molecules that act as axon guidance cues in the nervous system. Recent research showed increased expression of semaphorin 3C correlates with cancers that possess higher invasive and metastatic characteristics. For example, in breast cancer, inhibition of semaphorin 3C reduces adhesion and invasion. The aim of this study was to evaluate a possibility that semaphorin 3C might be a new prognostic marker in colorectal cancer. Material and methods: We used two cohorts. Cohort 1 was the 192 patients with colorectal cancer resected surgically between 2009 and 2010. We used GSE 14333 dataset as Cohort 2 which included 226 patients with the colorectal cancer. In each cohorts, we divided the patients in to the two groups i.e., high or low semaphorin 3C expression group using receiver operating characteristic (ROC) curve based on the information of the recurrence of colorectal cancer. Disease-free-survival (DFS) rates were calculated using Kaplan-Meier methods. Differences between curves were evaluated with the log-rank test. Results: In the cohort 1, the DFS in the patients with higher expression was worse than that in the patients with lower expression (P = 0.0467). Semaphorin 3C has two probes (203788_s_at and 203789_s_at) in GSE14333. In the cohort 2, using each two probe of semaphorin 3C, the DFS in the patients with higher expression was worse than that of those patients with lower expression (P = 0.0015 and P = 0.0003). Conclusions: This study suggested that Semaphorin 3C might be a new prognostic marker in colorectal cancer. Citation Format: Shingo Kawano, Kota Amemiya, Yuki Tsuchiya, Toshiaki Hagiwara, Hirokazu Matsuzawa, Yurika Makino, Shunsuke Motegi, Ryoichi Tsukamoto, Ryosuke Ichikawa, Kazumasa Kure, Kumpei Honjo, Hisashi Ro, Shingo Ito, Yu Okazawa, Rina Takahashi, Kosuke Mizukoshi, Masaya Kawai, Shinya Munakata, Koichiro Niwa, Shun Ishiyama, Kiichi Sugimoto, Hirohiko Kamiyama, Hiromitsu Komiyama, Makoto Takahashi, Yutaka Kojima, Michitoshi Goto, Atsushi Okuzawa, Yuichi Tomiki, Kazuhiro Sakamoto, Motohiro Kojima, Atsushi Ochiai. Semaphorin 3C has a possibility of a new prognostic marker in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2256. doi:10.1158/1538-7445.AM2017-2256

IA16: Current and emerging molecular diagnostic assays for colorectal cancer

Colorectal cancer is one of the most common types of cancer in the world. Several factors have been shown to put individuals at risk to CRC and these include age, the presence of polyps, inflammatory bowel disease, lifestyle, genetic background and family medical history. In the present study, we were related to the cancer antigen CEA and CA19-9 as prognostic markers in colorectal cancer. This may be helping us to detect colorectal cancer in an early stage. In this study, we have included 60 patients diagnosed with colorectal cancer. These patients were categorized as 10 patients between stage I-IV with disease dissemination and recurrence; 50 patients in between stage I- IV without dissemination and recurrence; 60 healthy controls. The cancer antigen CEA and Ca 19-9 were analyzed as prognostic markers in colorectal cancer. The statistical analysis was done by using SPSS software and expressed in terms of mean ± standard deviation. The p values (p˂0.005) were considered as statistically significant value. Person correlation between the cancer antigen CEA and CA-19-9 were analyzed. The cancer antigen CEA and Ca19-9 level were significantly increased in colorectal cancer patients as compared to the healthy controls group (p˂0.005). The significant Person correlation was observed in between CEA and CA19-9. The level of CEA and CA19-9 were significantly prognostic indicator in colorectal cancer. CEA and CA19-9 can be used as a diagnostic and prognostic marker for the screening of both colon and rectum cancer.

For many decades, cell-free nucleic acids have been known to be present in peripheral blood. Several studies have identified tumor-specific and/or tumor-associated alterations in the circulating nucleic acids of patients with various cancers. In recent years, cell-free microRNA (miRNA) have been stably detected in the plasma and serum, like other molecules; their presence in the blood has attracted the attention of researchers due to their potential use as valuable blood biomarkers.1Schwarzenbach H. Hoon D.S. Pantel K. Cell-free nucleic acids as biomarkers in cancer patients.Nat Rev Cancer. 2011; 11: 426-437Crossref PubMed Scopus (2199) Google Scholar MiRNAs are short, noncoding RNAs that play important roles in various physiologic and developmental processes. The mature miRNAs are produced from long primary transcripts through 2 sequential cleavage steps. The long primary miRNA transcript is cleaved by the Drosha complex in the nucleus, generating intermediate precursor miRNA. Precursor miRNA is transported by exportin-5 from the nucleus into the cytoplasm, and then subjected to further cleavage by a Dicer RNAase III enzyme, generating a short double-strand miRNA. One strand (guided strand) of mature miRNA is then incorporated into the RNA-induced silencing complex and subsequently hybridize to the 3′-untranslated region of their target mRNAs to repress translation or degrade these mRNAs. Thus, a single miRNA can influence the expression of hundreds of genes and allow them to function in a coordinated manner. Therefore, miRNAs have been implicated as key molecules in all cellular processes. Numerous studies have shown that alterations in miRNA expression correlate with various diseases, including the development and progression of cancer, and some miRNAs can function as oncogenes or tumor suppressors. These findings have opened up a new and interesting field in the diagnosis of cancer and the treatments of cancer patients. Mitchell et al2Mitchell P.S. Parkin R.K. Kroh E.M. et al.Circulating microRNAs as stable blood-based markers for cancer detection.Proc Natl Acad Sci U S A. 2008; 105: 10513-10518Crossref PubMed Scopus (6792) Google Scholar first demonstrated that circulating miRNAs had the potential to be new biomarkers in patients with solid cancers. In recent years, several papers have demonstrated that circulating miRNAs can also be detected in the peripheral blood of patients with digestive tract cancers. Although the origins and physiologic functions of cell-free miRNAs in the blood remain to be fully elucidated, a noninvasive assay for miRNAs should be developed to exploit these molecules as potential diagnostic and prognostic biomarkers. This assay undoubtedly contributes to an improvement in the clinical outcomes of cancer patients. In this article, we review the current state of biological and clinical research regarding circulating miRNAs of digestive tract cancer patients and discuss the future perspectives. It has been theorized that the necrosis and the apoptosis of tumor cells are the main sources of cell-free nucleic acids in the plasma and serum. However, several recent studies have demonstrated that extracellular nucleic acids, especially miRNAs, occur not only through cell lysis but also through active secretion.1Schwarzenbach H. Hoon D.S. Pantel K. Cell-free nucleic acids as biomarkers in cancer patients.Nat Rev Cancer. 2011; 11: 426-437Crossref PubMed Scopus (2199) Google Scholar, 3Valadi H. Ekström K. Bossios A. et al.Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells.Nat Cell Biol. 2007; 9: 654-659Crossref PubMed Scopus (9739) Google Scholar, 4Kosaka N. Iguchi H. Yoshioka Y. et al.Secretory mechanisms and intercellular transfer of microRNAs in living cells.J Biol Chem. 2010; 285: 17442-17452Abstract Full Text Full Text PDF PubMed Scopus (1591) Google Scholar, 5Pigati L. Yaddanapudi S.C. Iyengar R. et al.Selective release of microRNA species from normal and malignant mammary epithelial cells.PLoS One. 2010; 5: e13515Crossref PubMed Scopus (497) Google Scholar Cell-derived endogenous miRNAs are present in the blood in a remarkably stable form that is protected from endogenous RNase activity. In contrast, synthetic exogenous miRNAs are rapidly degraded when added directly to the plasma.2Mitchell P.S. Parkin R.K. Kroh E.M. et al.Circulating microRNAs as stable blood-based markers for cancer detection.Proc Natl Acad Sci U S A. 2008; 105: 10513-10518Crossref PubMed Scopus (6792) Google Scholar Kosaka et al4Kosaka N. Iguchi H. Yoshioka Y. et al.Secretory mechanisms and intercellular transfer of microRNAs in living cells.J Biol Chem. 2010; 285: 17442-17452Abstract Full Text Full Text PDF PubMed Scopus (1591) Google Scholar clearly demonstrated that a subset of miRNAs was packaged into exosome vesicles and released through a ceramide-dependent secretory mechanism. Arroyo et al6Arroyo J.D. Chevillet J.R. Kroh E.M. et al.Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma.Proc Natl Acad Sci U S A. 2011; 108: 5003-5008Crossref PubMed Scopus (2672) Google Scholar systematically investigated circulating miRNAs in the plasma and serum using differential centrifugation and size-exclusion chromatography techniques. This group demonstrated ≥2 populations of circulating miRNAs in the plasma and serum and discovered agonaute-2, a key effector protein involved in miRNA-mediated silencing as an miRNA carrier in the blood.6Arroyo J.D. Chevillet J.R. Kroh E.M. et al.Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma.Proc Natl Acad Sci U S A. 2011; 108: 5003-5008Crossref PubMed Scopus (2672) Google Scholar In addition, high-density lipoprotein has been described as an alternative transporter of extracellular miRNAs in human plasma.7Vickers K.C. Palmisano B.T. Shoucri B.M. et al.MicroRNAs are transported in plasma and delivered to recipient cells by high-density lipoproteins.Nat Cell Biol. 2011; 13: 423-433Crossref PubMed Scopus (2256) Google Scholar All circulating miRNAs, regardless of whether they are incorporated into protein complexes and/or cell-derived microvesicles, seem to be adequately protected against the degradation caused by the abundant RNases in human plasma and serum. Indeed, the extracellular miRNAs in the plasma and serum are extremely stable under severe conditions, such as extended storage and exposure to multiple freeze–thaw cycles.2Mitchell P.S. Parkin R.K. Kroh E.M. et al.Circulating microRNAs as stable blood-based markers for cancer detection.Proc Natl Acad Sci U S A. 2008; 105: 10513-10518Crossref PubMed Scopus (6792) Google Scholar Regarding the composition of circulating miRNAs, Pigati et al5Pigati L. Yaddanapudi S.C. Iyengar R. et al.Selective release of microRNA species from normal and malignant mammary epithelial cells.PLoS One. 2010; 5: e13515Crossref PubMed Scopus (497) Google Scholar investigated the difference between extracellular and cellular miRNAs using epithelial cell lines and concluded that the release of miRNAs did not necessarily reflect the abundance of miRNAs in the cell of origin. Kosaka et al4Kosaka N. Iguchi H. Yoshioka Y. et al.Secretory mechanisms and intercellular transfer of microRNAs in living cells.J Biol Chem. 2010; 285: 17442-17452Abstract Full Text Full Text PDF PubMed Scopus (1591) Google Scholar also demonstrated that some specific miRNAs were expressed to a greater extent in cell-derived exosomes compared with their donor cells.4Kosaka N. Iguchi H. Yoshioka Y. et al.Secretory mechanisms and intercellular transfer of microRNAs in living cells.J Biol Chem. 2010; 285: 17442-17452Abstract Full Text Full Text PDF PubMed Scopus (1591) Google Scholar Moreover, other groups demonstrated that the non–vesicle-associated miRNA profiles within protein complexes were distinctly different from the purified, exosomes-associated miRNA profiles.6Arroyo J.D. Chevillet J.R. Kroh E.M. et al.Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma.Proc Natl Acad Sci U S A. 2011; 108: 5003-5008Crossref PubMed Scopus (2672) Google Scholar These findings indicate that intracellular miRNAs are exported to the extracellular environment through a selective secretion mechanism. Interestingly, recent studies have demonstrated that extracellular miRNAs not only circulate in stable forms, but can also be incorporated into other surrounding and distant recipient cells in which they fulfill distinctive functions.8Skog J. Würdinger T. van Rijn S. et al.Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers.Nat Cell Biol. 2008; 10: 1470-1476Crossref PubMed Scopus (3954) Google Scholar, 9Rechavi O. Erlich Y. Amram H. et al.Cell contact–dependent acquisition of cellular and viral nonautonomously encoded small RNAs.Genes Dev. 2009; 23: 1971-1979Crossref PubMed Scopus (100) Google Scholar, 10Zernecke A. Bidzhekov K. Noels H. et al.Delivery of microRNA-126 by apoptotic bodies induces CXCL12-dependent vascular protection.Sci Signal. 2009; 2: ra81Crossref PubMed Scopus (1130) Google Scholar, 11Kosaka N. Iguchi H. Ochiya T. Circulating microRNA in body fluid: a new potential biomarker for cancer diagnosis and prognosis.Cancer Sci. 2010; 101: 2087-2092Crossref PubMed Scopus (1144) Google Scholar, 12Pegtel D.M. Cosmopoulos K. Thorley–Lawson D.A. et al.Functional delivery of viral miRNAs via exosomes.Proc Natl Acad Sci U S A. 2010; 107: 6328-6333Crossref PubMed Scopus (1309) Google Scholar, 13Kosaka N. Iguchi H. Yoshioka Y. et al.Competitive Interactions of Cancer Cells and Normal Cells via Secretory MicroRNAs.J Biol Chem. 2012; 287: 1397-1405Abstract Full Text Full Text PDF PubMed Scopus (235) Google Scholar Rechavi et al9Rechavi O. Erlich Y. Amram H. et al.Cell contact–dependent acquisition of cellular and viral nonautonomously encoded small RNAs.Genes Dev. 2009; 23: 1971-1979Crossref PubMed Scopus (100) Google Scholar demonstrated that functional signals spread across cell boundaries between immune cells in a contact-dependent manner. Pegtel et al12Pegtel D.M. Cosmopoulos K. Thorley–Lawson D.A. et al.Functional delivery of viral miRNAs via exosomes.Proc Natl Acad Sci U S A. 2010; 107: 6328-6333Crossref PubMed Scopus (1309) Google Scholar reported that Epstein–Barr virus miRNAs were secreted from infected B cells and were present in both the circulation and noninfected non-B cells. This group also demonstrated that miRNAs were transferred from infected to noninfected cells in vivo and were functional (upon transfer via exosomes) in primary monocyte-derived dendritic cells. Other groups have shown that miR-126 in apoptotic bodies derived from atherosclerotic endothelial cells induces the CXCL-12–dependent vascular protection process in recipient vascular cells.10Zernecke A. Bidzhekov K. Noels H. et al.Delivery of microRNA-126 by apoptotic bodies induces CXCL12-dependent vascular protection.Sci Signal. 2009; 2: ra81Crossref PubMed Scopus (1130) Google Scholar There have also been some reports regarding miRNA-mediated intercellular communication in a neoplastic environment. Skog et al8Skog J. Würdinger T. van Rijn S. et al.Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers.Nat Cell Biol. 2008; 10: 1470-1476Crossref PubMed Scopus (3954) Google Scholar reported that microvesicles that housed miRNAs derived from glioblastomas were taken up by and fulfilled functions in human brain microvascular endothelial cells in culture. Kosaka et al4Kosaka N. Iguchi H. Yoshioka Y. et al.Secretory mechanisms and intercellular transfer of microRNAs in living cells.J Biol Chem. 2010; 285: 17442-17452Abstract Full Text Full Text PDF PubMed Scopus (1591) Google Scholar also demonstrated that miR-146a, which is a tumor-suppressive miRNA in prostate cancer, significantly knocked down the target ROCK1 protein expression and decreased cell proliferation in a recipient prostate cancer cell line.4Kosaka N. Iguchi H. Yoshioka Y. et al.Secretory mechanisms and intercellular transfer of microRNAs in living cells.J Biol Chem. 2010; 285: 17442-17452Abstract Full Text Full Text PDF PubMed Scopus (1591) Google Scholar Their subsequent paper demonstrated that a variety of tumor-suppressive miRNAs were secreted by a normal adult prostatic epithelial cell line, and among these secretory miRNAs, miR-143 could inhibit growth exclusively in cancer cells both in vitro and in vivo.13Kosaka N. Iguchi H. Yoshioka Y. et al.Competitive Interactions of Cancer Cells and Normal Cells via Secretory MicroRNAs.J Biol Chem. 2012; 287: 1397-1405Abstract Full Text Full Text PDF PubMed Scopus (235) Google Scholar Other groups found that the let-7 miRNA family was abundant in the extracellular fractions derived from a metastatic gastric cancer (GC) cell line, but not those derived from a low metastatic parental cell line, and it has been speculated that some cancer cells maintain their oncogenesis via specific extracellular miRNAs.14Ohshima K. Inoue K. Fujiwara A. et al.Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line.PLoS One. 2010; 5: e13247Crossref PubMed Scopus (515) Google Scholar On the other hand, exosomes released from neoplastic cells have been reported to suppress immune surveillance, and cell-free miRNAs contained within the exosomes may be responsible for the immunosuppression systems.15Zhang H.G. Grizzle W.E. Exosomes and cancer: a newly described pathway of immune suppression.Clin Cancer Res. 2011; 17: 959-964Crossref PubMed Scopus (233) Google Scholar These findings support the presence of miRNA-mediated intercellular communication in the normal cellular environment and the tumor environment (Figure 1) . Several methods can be used for extracting miRNAs; however, efficient protocols with high reproducibility should be utilized for the extraction of circulating miRNAs owing to the small amounts present in the plasma and serum. Commercial extraction kits that utilize glass fiber filters in the purification process have been widely used for clinical blood samples, and there are several methods for quantification. A polymerase chain reaction (PCR)-based technique using a stem-loop reverse-transcriptase (RT) primer has been widely used for determining quantity. A microarray assay, which can analyze hundreds of miRNAs simultaneously, has also been utilized for the identification of a specific marker among many circulating miRNAs. Recent advances in technology allow for the use of an oligonucleotide array to quantify the amount of circulating miRNAs without the need for PCR. Most recently, researchers have identified circulating miRNAs as new diagnostic markers in patients with cancer using direct sequencing methods16Brase J.C. Wuttig D. Kuner R. et al.Serum microRNAs as non–invasive biomarkers for cancer.Mol Cancer. 2010; 9: 306Crossref PubMed Scopus (366) Google Scholar (Table 117Zhang C. Wang C. Chen X. et al.Expression profile of microRNAs in serum: a fingerprint for esophageal squamous cell carcinoma.Clin Chem. 2010; 56: 1871-1879Crossref PubMed Scopus (289) Google Scholar, 18Komatsu S. Ichikawa D. Takeshita H. et al.Circulating microRNAs in plasma of patients with oesophageal squamous cell carcinoma.Br J Cancer. 2011; 105: 104-111Crossref PubMed Scopus (233) Google Scholar, 19Tsujiura M. Ichikawa D. Komatsu S. et al.Circulating microRNAs in plasma of patients with gastric cancers.Br J Cancer. 2010; 102: 1174-1179Crossref PubMed Scopus (578) Google Scholar, 20Liu R. Zhang C. Hu Z. et al.A five-microRNA signature identified from genome–wide serum microRNA expression profiling serves as a fingerprint for gastric cancer diagnosis.Eur J Cancer. 2011; 47: 784-791Abstract Full Text Full Text PDF PubMed Scopus (392) Google Scholar, 21Liu H. Zhu L. Liu B. et al.Genome-wide microRNA profiles identify miR-–378 as a serum biomarker for early detection of gastric cancer.Cancer Lett. 2012; 316: 196-203Crossref PubMed Scopus (234) Google Scholar, 22Konishi H. Ichikawa D. Komatsu S. et al.Detection of gastric cancer–associated microRNAs on microRNA microarray comparing pre- and post-operative plasma.Br J Cancer. 2012; 106: 740-747Crossref PubMed Scopus (165) Google Scholar, 23Ng E.K. Chong W.W. Jin H. et al.Differential expression of microRNAs in plasma of patients with colorectal cancer: a potential marker for colorectal cancer screening.Gut. 2009; 58: 1375-1381Crossref PubMed Scopus (1055) Google Scholar, 24Huang Z. Huang D. Ni S. et al.Plasma microRNAs are promising novel biomarkers for early detection of colorectal cancer.Int J Cancer. 2010; 127: 118-126Crossref PubMed Scopus (868) Google Scholar, 25Wang L.G. Gu J. Serum is a promising novel marker for early detection of colorectal 2012; PubMed Scopus Google Scholar, H. Zhang L. et al.Circulating plasma is a novel biomarker for metastatic cancer and One. 2011; PubMed Scopus Google Scholar, Huang et al.Circulating directly from plasma is a potential diagnostic and prognostic marker of colorectal cancer and is with 2010; PubMed Scopus Google Scholar, Y. Kosaka N. M. et as a potential diagnostic marker for 2009; PubMed Scopus Google Scholar, Zhang K. H. et al.Circulating microRNAs as biomarkers for 2011; PubMed Scopus Google Scholar, Wang H. et al.Serum as for in with B One. 2011; PubMed Scopus Google Scholar, J. C. X. et al.Circulating and in patients with or 2011; PubMed Scopus Google Scholar, J. L. X. Hu J. et al.Plasma to 2011; PubMed Scopus Google Scholar, Y. H. H. et al.Circulating as a novel biomarker for 2012; 56: Full Text Full Text PDF PubMed Scopus Google Scholar, J. Y. X. et al.Serum microRNA as a potential marker for Sci. 2011; PubMed Scopus Google Scholar, J. Chen J. et al.MicroRNAs in plasma of patients as novel blood-based biomarkers of Res. 2009; 2: PubMed Scopus Google Scholar, A. N. et and and in with circulating and Res. 2010; PubMed Scopus Google Scholar, Huang X. H. et al.Circulating as a novel marker in 2010; PubMed Scopus Google Scholar, S. K. Chen et expressed miRNAs in the plasma may provide a signature for J Res. 2010; Google Scholar, N. A. et and serum microRNAs in the and in patients with One. 2011; PubMed Scopus Google Scholar, R. Komatsu S. Ichikawa D. et diagnostic of circulating in plasma of patients with J Cancer. 2011; 105: PubMed Scopus Google Scholar, J. J. Y. et of plasma microRNAs with serum for early detection of cancer.Int J Cancer. 2011; Scopus Google miRNAs in cell or other C. Wang C. Chen X. et al.Expression profile of microRNAs in serum: a fingerprint for esophageal squamous cell carcinoma.Clin Chem. 2010; 56: 1871-1879Crossref PubMed Scopus (289) Google S. Ichikawa D. Takeshita H. et al.Circulating microRNAs in plasma of patients with oesophageal squamous cell carcinoma.Br J Cancer. 2011; 105: 104-111Crossref PubMed Scopus (233) Google M. Ichikawa D. Komatsu S. et al.Circulating microRNAs in plasma of patients with gastric cancers.Br J Cancer. 2010; 102: 1174-1179Crossref PubMed Scopus (578) Google or other R. Zhang C. Hu Z. et al.A five-microRNA signature identified from genome–wide serum microRNA expression profiling serves as a fingerprint for gastric cancer diagnosis.Eur J Cancer. 2011; 47: 784-791Abstract Full Text Full Text PDF PubMed Scopus (392) Google in the H. Zhu L. Liu B. et al.Genome-wide microRNA profiles identify miR-–378 as a serum biomarker for early detection of gastric cancer.Cancer Lett. 2012; 316: 196-203Crossref PubMed Scopus (234) Google H. Ichikawa D. Komatsu S. et al.Detection of gastric cancer–associated microRNAs on microRNA microarray comparing pre- and post-operative plasma.Br J Cancer. 2012; 106: 740-747Crossref PubMed Scopus (165) Google E.K. Chong W.W. Jin H. et al.Differential expression of microRNAs in plasma of patients with colorectal cancer: a potential marker for colorectal cancer screening.Gut. 2009; 58: 1375-1381Crossref PubMed Scopus (1055) Google Z. Huang D. Ni S. et al.Plasma microRNAs are promising novel biomarkers for early detection of colorectal cancer.Int J Cancer. 2010; 127: 118-126Crossref PubMed Scopus (868) Google in the L.G. Gu J. Serum is a promising novel marker for early detection of colorectal 2012; PubMed Scopus Google cancer H. Zhang L. et al.Circulating plasma is a novel biomarker for metastatic cancer and One. 2011; PubMed Scopus Google in the Huang et al.Circulating directly from plasma is a potential diagnostic and prognostic marker of colorectal cancer and is with 2010; PubMed Scopus Google in the Y. Kosaka N. M. et as a potential diagnostic marker for 2009; PubMed Scopus Google Zhang K. H. et al.Circulating microRNAs as biomarkers for 2011; PubMed Scopus Google Wang H. et al.Serum as for in with B One. 2011; PubMed Scopus Google not for J. C. X. et al.Circulating and in patients with or 2011; PubMed Scopus Google in the J. L. X. Hu J. et al.Plasma to 2011; PubMed Scopus Google Y. H. H. et al.Circulating as a novel biomarker for 2012; 56: Full Text Full Text PDF PubMed Scopus Google J. Y. X. et al.Serum microRNA as a potential marker for Sci. 2011; PubMed Scopus Google in the J. Chen J. et al.MicroRNAs in plasma of patients as novel blood-based biomarkers of Res. 2009; 2: PubMed Scopus Google A. N. et and and in with circulating and Res. 2010; PubMed Scopus Google in the Huang X. H. et al.Circulating as a novel marker in 2010; PubMed Scopus Google S. K. Chen et expressed miRNAs in the plasma may provide a signature for J Res. 2010; Google N. A. et and serum microRNAs in the and in patients with One. 2011; PubMed Scopus Google R. Komatsu S. Ichikawa D. et diagnostic of circulating in plasma of patients with J Cancer. 2011; 105: PubMed Scopus Google in the J. J. Y. et of plasma microRNAs with serum for early detection of cancer.Int J Cancer. 2011; Scopus Google microRNA colorectal direct esophageal gastric polymerase chain in the in a new microRNA colorectal direct esophageal gastric polymerase chain Zhang et C. Wang C. Chen X. et al.Expression profile of microRNAs in serum: a fingerprint for esophageal squamous cell carcinoma.Clin Chem. 2010; 56: 1871-1879Crossref PubMed Scopus (289) Google Scholar have investigated the serum miRNA profiles of patients with esophageal squamous cell using miRNAs using direct this group identified serum miRNAs and as biomarkers. The under the for the miRNAs were for serum tumor and patients in the early of the could be from using the miRNA C. Wang C. Chen X. et al.Expression profile of microRNAs in serum: a fingerprint for esophageal squamous cell carcinoma.Clin Chem. 2010; 56: 1871-1879Crossref PubMed Scopus (289) Google Scholar plasma from group also the plasma expression of miRNAs that were to be with the development of on found that the plasma of to be in patients in the and the were significantly in compared with A of the in plasma was the of an in serum tumor which that circulating miRNAs may be for diagnosis of in S. Ichikawa D. Takeshita H. et al.Circulating microRNAs in plasma of patients with oesophageal squamous cell carcinoma.Br J Cancer. 2011; 105: 104-111Crossref PubMed Scopus (233) Google Scholar However, there have been reports regarding circulating miRNAs for the other group first reported the of circulating miRNAs as biomarkers in patients with miRNAs and which have been reported to be in as miRNAs and their in plasma using In the plasma of these miRNAs the tumor miRNA and were significantly in patients in the also found that the plasma of these miRNAs were significantly in compared with M. Ichikawa D. Komatsu S. et al.Circulating microRNAs in plasma of patients with gastric cancers.Br J Cancer. 2010; 102: 1174-1179Crossref PubMed Scopus (578) Google Scholar Liu et R. Zhang C. Hu Z. et al.A five-microRNA signature identified from genome–wide serum microRNA expression profiling serves as a fingerprint for gastric cancer diagnosis.Eur J Cancer. 2011; 47: 784-791Abstract Full Text Full Text PDF PubMed Scopus (392) Google Scholar have investigated the serum miRNA profiles of patients with using miRNAs using direct this group identified serum miRNAs and as biomarkers for found that the expression of serum miRNAs was with the tumor and the for the miRNAs were for serum tumor R. Zhang C. Hu Z. et al.A five-microRNA signature identified from genome–wide serum microRNA expression profiling serves as a fingerprint for gastric cancer diagnosis.Eur J Cancer. 2011; 47: 784-791Abstract Full Text Full Text PDF PubMed Scopus (392) Google Scholar Other groups have investigated serum miRNA profiles using miRNA and concluded that was the biomarker for detection in the serum, with and the expression in primary was compared with normal H. Zhu L. Liu B. et al.Genome-wide microRNA profiles identify miR-–378 as a serum biomarker for early detection of gastric cancer.Cancer Lett. 2012; 316: 196-203Crossref PubMed Scopus (234) Google Scholar The selective release of specific cellular miRNAs from the tumor cells or from normal as could these et E.K. Chong W.W. Jin H. et al.Differential expression of microRNAs in plasma of patients with colorectal cancer: a potential marker for colorectal cancer screening.Gut. 2009; 58: 1375-1381Crossref PubMed Scopus (1055) Google Scholar first reported that circulating miRNAs could be potential noninvasive markers for colorectal cancer patients. This group plasma from patients using a miRNA microarray and found that and were in the plasma of patients and in compared with they demonstrated that the detection of could from other and The was with and an E.K. Chong W.W. Jin H. et al.Differential expression of microRNAs in plasma of patients with colorectal cancer: a potential marker for colorectal cancer screening.Gut. 2009; 58: 1375-1381Crossref PubMed Scopus (1055) Google this has been by other Huang et Z. Huang D. Ni S. et al.Plasma microRNAs are promising novel biomarkers for early detection of colorectal cancer.Int J Cancer. 2010; 127: 118-126Crossref PubMed Scopus (868) Google Scholar the plasma of miRNAs that were to be in in This group found that and had a diagnostic for and and that patients with could be from the using these 2 Z. Huang D. Ni S. et al.Plasma microRNAs are promising novel biomarkers for early detection of colorectal cancer.Int J Cancer. 2010; 127: 118-126Crossref PubMed Scopus (868) Google Scholar Wang et L.G. Gu J. Serum is a promising novel marker for early detection of colorectal 2012; PubMed Scopus Google Scholar the serum expression of miRNAs and in and metastatic patients and reported that serum had the potential to be a noninvasive biomarker for the early detection of in L.G. Gu J. Serum is a promising novel marker for early detection of colorectal 2012; PubMed Scopus Google Scholar et Huang et al.Circulating directly from plasma is a potential diagnostic and prognostic marker of colorectal cancer and is with 2010; PubMed Scopus Google Scholar reported the first of the direct of circulating miRNAs from the plasma without RNA extraction and found that the direct of plasma could be used as a potential noninvasive diagnostic and prognostic biomarker for Huang et al.Circulating directly from plasma is a potential diagnostic and prognostic marker of colorectal cancer and is with 2010; PubMed Scopus Google Scholar et Y. Kosaka N. M. et as a potential diagnostic marker for 2009; PubMed Scopus Google Scholar first reported the of circulating miRNAs as biomarker for cancer identified as an miRNA of by miRNA profiles in the process of development using of serum was found in the of of patients and the were significantly Y. Kosaka N. M. et as a potential diagnostic marker for 2009; PubMed Scopus Google Scholar et Wang H. et al.Serum as for in with B One. 2011; PubMed Scopus Google Scholar investigated the serum of miRNAs and to identify new but only the was significantly in the serum of patients compared with or viral B patients without and the of were also significantly in serum Wang H. et al.Serum as for in with B One. 2011; PubMed Scopus Google Scholar et J. L. X. Hu J. et al.Plasma to 2011; PubMed Scopus Google Scholar investigated plasma miRNAs using an miRNA microarray in a of patients. This group first on the of the microarray and subsequently identified miRNAs and using for the miRNA reported that their miRNA could patients from B patients and patients and it is a promising marker for the early diagnosis of J. L. X. Hu J. et al.Plasma to 2011; PubMed Scopus Google Scholar Wang et J. Chen J. et al.MicroRNAs in plasma of patients as novel blood-based biomarkers of Res. 2009; 2: PubMed Scopus Google Scholar the plasma of miRNAs and that have been implicated in cancer development in The plasma of miRNA were in patients compared with the The for miRNA were not but they to when the miRNAs were J. Chen J. et al.MicroRNAs in plasma of patients as novel blood-based biomarkers of Res. 2009; 2: PubMed Scopus Google Scholar et A. N. et and and in with circulating and Res. 2010; PubMed Scopus Google Scholar identified 2 of the family and that were in using This group also found that these 2 miRNAs were significantly in the of and patients compared with of and et N. A. et and serum microRNAs in the and in patients with One. 2011; PubMed Scopus Google Scholar identified serum miRNAs and as diagnostic biomarkers of in an circulating miRNAs and demonstrated that the circulating miRNAs could as not only diagnostic markers but also of the Liu et J. J. Y. et of plasma microRNAs with serum for early detection of cancer.Int J Cancer. 2011; Scopus Google Scholar miRNA and in the plasma of and patients using All miRNA were significantly in patients compared with patients and This group concluded that the of and the marker was among various in from and J. J. Y. et of plasma microRNAs with serum for early detection of cancer.Int J Cancer. 2011; Scopus Google Scholar Circulating miRNAs are for use as valuable biomarkers owing to their in the and of Therefore, circulating miRNAs may be used for the clinical in cancer and early or and for the the small of which is a that miRNAs from other for an of using and direct

Nuclear S100A4 is a novel prognostic marker in colorectal cancer

Background and Aims: Current staging classifications in colorectal cancer are not able to accurately predict patient outcome, and the need for novel prognostic markers is evident. S100A4 is a Ca2+-binding protein which promotes metastasis in several tumor types, and the aim of the present study was to investigate the prognostic impact of S100A4 expression in colorectal cancer. Methods: 242 patients with curatively resected adenocarcinoma of the colon or rectum were prospectively included in the study at the time of surgery. S100A4 expression was analyzed by immunohistochemistry, and associations with clinicopathological variables and patient outcome were investigated. Results: Nuclear expression of S100A4 was observed in 29% and cytoplasmic expression was observed in 64% of the tumors. In univariate analysis, nuclear S100A4 was a significant predictor of metastasis-free (P= .006) and overall survival (P= .01), whereas cytoplasmic S100A4 was not associated with patient outcome. In multivariate analysis, nuclear localization was associated with metastasis-free (P= .03) and overall survival (P= .02). Interestingly, the prognostic impact was largely confined to TNM stage II, and stage II patients with tumors expressing nuclear S100A4 had a similar prognosis as stage III patients. Conclusions: Nuclear expression of S100A4 is a novel prognostic marker in colorectal cancer, and the prognostic value in TNM stage II suggests that nuclear S100A4 could be used in the stratification of stage II patients for adjuvant treatment. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-73.

Colorectal cancer (CRC) is currently the third and the second most common cancer in men and in women, respectively. Every year, more than one million new CRC cases and more than half a million deaths are reported worldwide. The majority of new cases occur in developed countries. Current screening methods have significant limitations. Therefore, a lot of scientific effort is put into the development of new diagnostic biomarkers of CRC. Currently used prognostic markers are also limited in assessing the effectiveness of CRC therapy. MicroRNAs (miRNAs) are a promising subject of research especially since single miRNA can recognize a variety of different mRNA transcripts. MiRNAs have important roles in epigenetic regulation of basic cellular processes, such as proliferation, apoptosis, differentiation, and migration, and may serve as potential oncogenes or tumor suppressors during cancer development. Indeed, in a large variety of human tumors, including CRC, significant distortions in miRNA expression profiles have been observed. Thus, the use of miRNAs as diagnostic and prognostic biomarkers in cancer, particularly in CRC, appears to be an inevitable consequence of the advancement in oncology and gastroenterology. Here, we review the literature to discuss the potential usefulness of selected miRNAs as diagnostic and prognostic biomarkers in CRC.

AimThe aim of this study is to examine the role of the microrchidia (MORC) family, a group of chromatin remodeling proteins, as the therapeutic and prognostic markers for colorectal cancer (CRC).BackgroundMORC protein family genes are a highly conserved nucleoprotein superfamily whose members share a common domain but have distinct biological functions. Previous studies have analyzed the roles of MORCs as epigenetic regulators and chromatin remodulators; however, the involvement of MORCs in the development and pathogenesis of CRC was less examined.ObjectiveThe current work examined the role of the MORCs as the therapeutic and prognostic markers for CRC.MethodsThe expressions and prognostic significance of MORC family genes in CRC were explored. The role of these genes in tumor immunity was comprehensively analyzed in terms of their functions in immune cell infiltration, tumor microenvironment (TME), and their interaction with immune regulatory genes such as immunosuppressive genes, immune checkpoints and immunostimulatory genes. The relations between MORC family genes, tumor mutation burden (TMB), DNA, mismatch repair (MMR), RNA methylation, microsatellite instability (MSI), and drug sensitivity were investigated using the R statistical software. The expressions of MORC4 in 150 CRC tissues and 60 paracancer tissues were detected by immunohistochemical method. CRC cell proliferation, migration, and invasion were measured by cell counting kit-8 (CCK-8), scratch assay, and transwell cell invasion assay.ResultsThe expressions of MORC2 and MORC4 were significantly upregulated, whereas those of MORC1 and MORC3 were noticeably downregulated in CRC in comparison to their expressions in normal colorectal mucosal tissues. Patients with high-expressed MORC2 showed a more unfavorable prognosis than those with a low MORC2 level. Functional annotation analysis identified 100 MORC family genes with the most significant negative or positive correlations to diabetic cardiomyopathy, amyotrophic lateral sclerosis, oxidative phosphorylation, Huntington’s disease, thermogenesis, Parkinson’s disease, olfactory transduction, Alzheimer’s disease, prion disease. MORC3 expression was positively correlated with Stromal score, Immune score and ESTIMATE score, while MORC2 expression was negatively related to the three scores in CRC, these correlations were not statistically significant. Additionally, the MORC family genes were significantly positively correlated with tumor-infiltrating immune cells such as T helper cells and exhibited close relations to some immunosuppressive genes such as CXCR4 and PVR, immunostimulatory genes such as TGFBR1, KDR, and CD160 as well as some immune checkpoint genes. It was found that the expressions of some members of MORC family genes were positively correlated with DNA methylation, MSI, TMB, MMRs, and drug sensitivity in CRC and that the mRNA and protein levels of MORC4 were remarkably upregulated in CRC tissues than in adjacent normal tissues (P<0.05). In the MORC4 knockdown group, DLD-1 cell proliferation was more inhibited than in the negative control (NC) and siRNA groups (P<0.05). Furthermore, the migratory capacity of DLD-1 cells and the number of cells crossing the basement membrane in the MORC4 knockdown group were reduced compared to the NC and siRNA groups (all P<0.05).ConclusionThe expressions of MORC family genes were significantly different in CRC samples, which was related to the immune cell infiltration and prognosis of CRC. Thus, the MORC family genes were considered as markers for indicating the clinical immunotherapy and prognostic outcome of CRC.

Although various studies have demonstrated that growth differentiation factor 15 (GDF15) might be a potential diagnostic and prognostic marker in colorectal cancer (CRC) patients, the results are inconsistent and the statistical power of individual studies is also insufficient. An original study was conducted to explore the diagnostic and prognostic value of serum GDF15 in CRC patients. We also conducted a meta‐analysis study which aimed to summarize the diagnostic and prognostic performance of serum GDF15 in CRC. We searched PubMed and ISI Web of Knowledge up to 1 November 2014 for eligible studies. In order to explore the diagnostic performance of GDF15, standardized mean difference (SMD) and their 95% confidence intervals (CI) were estimated and receiver‐operating characteristic (ROC) curves were constructed. For prognostic meta‐analysis, study‐specific hazard ratios (HRs) of serum GDF15 for survival were summarized. A total of eight studies were included in the meta‐analyses. Our results revealed that serum GDF15 levels in CRC patients were higher than those in healthy controls (SMD = 1.08, 95% CI: 0.56–1.59, P < 0.001). For discriminating CRC from healthy controls, the AUC of GDF15 was 0.816 (95% CI: 0.792–0.838). The sensitivity and specificity were 58.9% (95% CI: 55.0–62.8) and 92.08% (95% CI: 89.2–94.4), respectively, when a cut‐off value of 1099 pg/ml was established. Besides, higher GDF15 expression level was associated with worse overall survival for CRC patients (pooled HR = 2.09, 95% CI: 1.47–2.96). In conclusion, the present meta‐analysis suggests that serum GDF15 may be a useful diagnostic and prognostic biomarker for CRC.

Colorectal cancer (CRC) has a complex pathogenesis involving multiple sequential steps with accumulation of genetic alterations including mutations, gene amplification, and epigenetic changes. Treatment of CRC has undergone a paradigm shift over the past decade due in part to a better understanding of the biology of the disease and development of newer drugs including biologic agents. In the era of personalized medicine, it is attractive to investigate the molecular pathways leading to colorectal cancer tumorigenesis, thus raising the possibility of identifying novel therapeutic targets. It has intuitive appeal to hypothesize that biomarkers that have prognostic and/or predictive value are those that are intimately connected to the pathogenesis of CRC. In this article, we focus on prognostic and predictive markers in CRC that have a substantial body of data in support of their potential role in routine clinical practice.

Colorectal cancer is one of the most common leading causes of death worldwide. Prognostic at an early stage is a useful way that decrease and avoid mortality. Although remarkable progress has been made to investigate the underlying mechanism, the understanding of the complicated carcinogenesis process was enormously hindered by large-scale tumor heterogeneity. Here we proposed that the prognosis-related gene THBS2, responsible for cooperativity disorientation, probably contain untapped prognostic resource of colorectal cancer. We originally established Spearman correlation transition, Kaplan–Meier survival analysis and meta-analysis that combine public dataset and clinical samples to quantify the prognostic value of THBS2. THBS2 could be considered as a novel prognostic marker in colorectal cancer.

Ubiquinol cytochrome c reductase binding protein (UQCRB) is important for mitochondrial complex III stability, electron transport, cellular oxygen sensing and angiogenesis. However, its potential as a prognostic marker in colorectal cancer (CRC) remains unclear. The aim of this study was to determine whether UQCRB can be used as a diagnostic molecular marker for CRC. The correlation between the expression of three genes (UQCRB, UQCRFS1 and MT-CYB) in the mitochondrial respiratory chain complex III and clinico-pathological features was determined. Compared to non-tumor tissues, UQCRB gene expression was upregulated in CRC tissues. Gene and protein expression of the genes were positively correlated. Copy number variation (CNV) differences in UQCRB were observed in CRC tissues (1.32-fold) compared to non-tumor tissues. The CNV of UQCRB in CRC tissues increased proportionally with gene expression and clinical stage. Single-nucleotide polymorphisms in the 3′-untranslated region of UQCRB (rs7836698 and rs10504961) were investigated, and the rs7836698 polymorphism was associated with CRC clinical stage. DNA methylation of the UQCRB promoter revealed that most CRC patients had high methylation levels (12/15 patients) in CRC tissues compared to non-tumor tissues. UQCRB overexpression and CNV gain were correlated with specific CRC clinico-pathological features, indicating clinical significance as a prognostic predictor in CRC. Gene structural factors may be more important than gene transcription repression factors with respect to DNA methylation in UQCRB overexpression. Our results provide novel insights into the critical role of UQCRB in regulating CRC, supporting UQCRB as a new candidate for the development of diagnostics for CRC patients.

BACKGROUNDAs a novel endogenous anti-angiogenic molecule, vasohibin 1 (VASH1) is not only expressed in tumor stroma, but also in tumor tissue. Moreover, studies have shown that VASH1 may be a prognostic marker in colorectal cancer (CRC). Knockdown of VASH1 enhanced transforming growth factor-β1 (TGF-β1)/Smad3 pathway activity and type I/III collagen production. Our previous findings suggest that ELL-associated factor 2 (EAF2) may play a tumor suppressor and protective role in the development and progression of CRC by regulating signal transducer and activator of transcription 3 (STAT3)/TGF-β1 signaling pathway. However, the functional role and mechanism of VASH1-mediated TGF-β1 related pathway in CRC has not been elucidated.AIMTo investigate the expression of VASH1 in CRC and its correlation with the expression of EAF2. Furthermore, we studied the functional role and mechanism of VASH1 involved in the regulation and protection of EAF2 in CRC cells in vitro.METHODSWe collected colorectal adenocarcinoma and corresponding adjacent tissues to investigate the clinical expression of EAF2 protein and VASH1 protein in patients with advanced CRC. Following, we investigated the effect and mechanism of EAF2 and VASH1 on the invasion, migration and angiogenesis of CRC cells in vitro using plasmid transfection.RESULTSOur findings indicated that EAF2 was down-regulated and VASH1 was up-regulated in advanced CRC tissue compared to normal colorectal tissue. Kaplan-Meier survival analysis showed that the higher EAF2 Level group and the lower VASH1 Level group had a higher survival rate. Overexpression of EAF2 might inhibit the activity of STAT3/TGF-β1 pathway by up-regulating the expression of VASH1, and then weaken the invasion, migration and angiogenesis of CRC cells.CONCLUSIONThis study suggests that EAF2 and VASH1 may serve as new diagnostic and prognostic markers for CRC, and provide a clinical basis for exploring new biomarkers for CRC. This study complements the mechanism of EAF2 in CRC cells, enriches the role and mechanism of CRC cell-derived VASH1, and provides a new possible subtype of CRC as a therapeutic target of STAT3/TGF-β1 pathway.

The intrinsic or mitochondrial apoptosis pathway is controlled by the interaction of antiapoptotic and pro-apoptotic members of the BCL-2 protein family. Activation of this death pathway plays a crucial role in cancer progression and chemotherapy responses. The BCL-2-related ovarian killer (BOK) possesses three BCL-2 homology domains and has been proposed to act in a similar pro-apoptotic pathway as the pro-apoptotic proteins BAX and BAK. In this study, we showed that stage II and III colorectal cancer patients possessed decreased levels of BOK protein in their tumours compared to matched normal tissue. BOK protein levels in tumours were also prognostic of clinical outcome but increased BOK protein levels surprisingly associated with earlier disease recurrence and reduced overall survival. We found no significant association of BOK protein tumour levels with ER stress markers GRP78 or GRP94 or with cleaved caspase-3. In contrast, BOK protein levels correlated with Calreticulin. These data indicate BOK as a prognostic marker in colorectal cancer and suggest that different activities of BOK may contribute to cancer progression and prognosis.

A Meta-Analysis of SMAD4 Immunohistochemistry as a Prognostic Marker in Colorectal Cancer