Novel Biomarker For Colorectal Cancer Research Articles

Identification of Novel Biomarkers for Metastatic Colorectal Cancer Using Angiogenesis-Antibody Array and Intracellular Signaling Array

Colorectal cancer (CRC) is one of the three leading causes for cancer mortality. CRC kills over 600,000 people annually worldwide. The most common cause of death from CRC is the metastasis to distant organs. However, biomarkers for CRC metastasis remain ill-defined. We compared primary and metastatic CRC cell lines for their angiogenesis-protein profiles and intracellular signaling profiles to identify novel biomarkers for CRC metastasis. To this end, we used primary and metastatic CRC cell lines as a model system and normal human colon cell line as a control. The angiogenesis profiles two isogenic CRC cell lines, SW480 and SW620, and HT-29 and T84 revealed that VEGF was upregulated in both SW620 and T84 whereas coagulation factor III, IGFBP-3, DPP IV, PDGF AA/AB, endothelin I and CXCL16 were downregulated specifically in metastatic cell lines. Furthermore, we found that TIMP-1, amphiregulin, endostatin, angiogenin were upregulated in SW620 whereas downregulated in T84. Angiogenin was downregulated in T84 and GM-CSF was also downregulated in SW620. To induce CRC cell metastasis, we treated cells with pro-inflammatory cytokine IL-6. Upon IL-6 treatment, epithelial-mesenchymal transition was induced in CRC cells. When DLD-1 and HT-29 cells were treated with IL-6; Akt, STAT3, AMPKα and Bad phosphorylation levels were increased. Interestingly, SW620 showed the same signal activation pattern with IL-6 treatment of HT-29 and DLD-1. Our data suggest that Akt, STAT3, AMPKα and Bad activation can be biomarkers for metastatic colorectal cancer. IL-6 treatment specifically reduced phosphorylation levels of EGFR, HER2 receptor, Insulin R and IGF-1R in receptor tyrosine kinase array study with HT-29. Taken together, we have identified novel biomarkers for metastatic CRC through the angiogenesis-antibody array and intracellular signaling array studies. Present study suggests that those novel biomarkers can be used as CRC prognosis biomarkers, and as potential targets for the metastatic CRC therapy.

Abstract 160: ZFAS1, long non-coding RNA with significance in colorectal cancer

Abstract Introduction: Colorectal cancer (CRC) is one of the most common cancers in the world. However, the existence of multiple known carcinogens and varying genetic backgrounds makes it difficult to determine which factors are most important in the development of CRC. Therefore, the identification of a bona fide molecule involved in progression of CRC has been greatly sought after. Long noncoding RNAs (lncRNA) are emerging as key molecules in human cancer, and some of them were shown to have potential for cancer diagnosis and prognosis. ZFAS1, an lncRNA, has been recently revealed involving in human breast cancer as putative tumor suppressor. However, whether ZFAS1 is involved in CRC pathogenesis or could serve as novel biomarker in CRC is unknown. Patients and methods: By use of high throughput technology we identified ZFAS1 as one of the highly up-regulated lncRNA in CRC tissue. Thus, aim of the current study was to explore the correlation between ZFAS1 expression and progression of CRC and its clinical significance. The expression levels of ZFAS1 were determined by quantitative real-time PCR and in selected cases by Northern blot and evaluated in larger cohort (120 cases) of CRC tissues and cell lines (HCT116, DLD1, SW620), which were compared to paired adjacent normal tissues or normal colonic cell lines. The potential relationship between ZFAS1 levels in tumor tissues and the clinicopathological features of CRC was also evaluated. Further, we assessed whether ZFAS1 influences cell proliferation, apoptosis, cell cycle and migration in-vitro. The different isoforms of ZFAS1 were determined by RACE-PCR and protein interacting partners of ZFAS1 were analyzed by pull-down experiment and mass spectrometry. The identified candidate proteins were evaluated by RNA-IP. Results: Our results showed that ZFAS1 expression in CRC tissues and cell lines were significantly increased compared with matched normal tissues (p < 0.0001). In addition, siRNA-mediated knockdown of ZFAS1 expression significantly influenced the proliferation of CRC cells (p < 0.05) Interestingly, ZFAS1 intronically hosts three C/D box snoRNAs (SNORDs): SNORD12, 12B, and 12B. The general assumption that noncoding SNORD-host transcripts function only as vehicles to generate snoRNAs, knockdown of ZFAS1 in a CRC cell lines resulted in decreased cellular proliferation, while not substantially altering the levels of the SNORDs. Conclusion: Our current study demonstrates that ZFAS1 levels are increased in CRC tissue and influences significantly proliferation of CRC cells in vitro, indicating its potential oncogenic functioning in CRC. The work has been supported by Project CZ.1.07/2.3.00/30.0037, and IGA MZCR NT13549-4/2012, NT13860-4/2012. Citation Format: Nithyananda Thorenoor, Petra Vychytilova, Jitka Mlcochova, Sonja Hombach, Markus Kretz, Jiri Sana, Ondrej Slaby. ZFAS1, long non-coding RNA with significance in colorectal cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 160. doi:10.1158/1538-7445.AM2015-160

Characterization of SLC22A18 as a tumor suppressor and novel biomarker in colorectal cancer

SLC22A18, solute carrier family 22, member 18, has been proposed to function as a tumor suppressor based on its chromosomal location at 11p15.5, mutations and aberrant splicing in several types of cancer and down-regulation in glioblastoma. In this study, we sought to demonstrate the significance of SLC22A18 as a tumor suppressor in colorectal cancer (CRC) and provide mechanistic bases for its function. We first showed that the expression of SLC22A18 is significantly down-regulated in tumor tissues using matched normal-tumor samples from CRC patients. This finding was also supported by publically accessible data from The Cancer Genome Atlas (TCGA). Functionally, SLC22A18 inhibits colony formation and induces of G2/M arrest consistent with being a tumor suppressor. Interestingly, suppression of KRAS by RNA interference promotes SLC22A18 expression, and expression of SLC22A18 in turn inhibits KRAS(G12D)-mediated anchorage independent growth of NIH3T3 cells indicating a mutual negative interaction. Finally, we evaluated diagnostic and prognostic values of SLC22A18 using clinical and gene expression data from TCGA which revealed a significantly worse long-term prognosis for patients with low level SLC22A18 expression. In sum, we established SLC22A18 as a tumor suppressor in colon epithelial cells and propose that SLC22A18 is potentially a marker of diagnostic and prognostic values.

Circulating miR-103 and miR-720 as novel serum biomarkers for patients with colorectal cancer.

Circulating microRNAs (miRNAs) have been reported as a biomarker for human malignancies, including colorectal cancer (CRC). The purpose of this study was to identify a novel biomarker for CRC through examination of serum miRNAs from the patients with CRC. Microarray analysis of miRNA expression was performed using paired pre- and post-operative serum from 10CRC patients. miR-103 and miR-720 decreased significantly in the post-operative serum when compared to pre-operative serum. With an extended scale validation by qRT-PCR (quantitative real-time polymerase chain reaction) in 30CRC patients, we confirmed that serum miR-103 and miR-720 decreased significantly after surgery (P=0.0004, and P=0.0274, respectively). Next, we examined serum miR-103 and miR-720 levels in 32 non-cancer patients and 84CRC patients, and we found that expression of these two miRNAs was significantly higher in CRC patients than non-cancer patients. Furthermore, clinical and pathological survey indicated that high expression of miR-103 was significantly associated with histological differentiation grade, and lymphatic invasion and high expression of miR-720 was significantly associated with male gender and lymph node metastasis. Our data suggest that circulating miR-103 and miR-720 show potential as novel serum biomarkers for CRC.

Methylated vimentin (mVim) DNA in blood as a novel biomarker in colorectal cancer: A retrospective study.

e14607 Background: Carcinoembryonic antigen (CEA) is a commonly used tumor marker in colorectal cancer (CRC) but has poor sensitivity (59%) and specificity (84%). Vimentin exon-1 sequences are hype...

大腸癌における新規バイオマーカーとしての血清ラミニン γ 2単鎖測定法の開発

大腸癌における新規バイオマーカーとしての血清ラミニン γ 2単鎖測定法の開発

The effect of colonoscopy on whole blood gene expression profile: an experimental investigation for colorectal cancer biomarker discovery.

Gene expression profiling of whole blood is showing great promise for the discovery of novel biomarkers for colorectal cancer (CRC) detection. Given the relatively low incidence rate of CRC in the general population, most blood samples collected prior to a colonoscopy were confirmed to be noncancerous afterward. Previous studies have relied on blood samples collected after a colonoscopy to reach the sufficient number of CRC cases. The present study aimed to determine the colonoscopy-induced variability in the blood transcriptome and its potential impact on biomarker discovery. Whole blood gene expression profiling was conducted using Affymetrix HG-U133Plus2 arrays. We analyzed 20 paired blood samples collected from healthy controls before and after colonoscopy, and 20 blood samples collected from CRC patients after colonoscopy. Utilizing hierarchical clustering analysis, samples collected before and after colonoscopy from the same subjects were closely clustered together, suggesting that the blood gene expression profiles primarily reflect the heterogeneity within each individual. A total of 914 genes were differentially expressed between controls and CRC patients, while gene expression did not differ between samples collected before and after colonoscopy. Using real-time PCR technology, we further validated six genes from published biomarkers in this study. Our results confirmed that the biomarkers identified in previous studies were not likely to be biased due to the colonoscopy effect. Our results showed that the colonoscopy-induced variations were minute compared to individuals' heterogeneity and disease-induced variations. These findings may serve as a basis for future development of blood-based transcriptomic biomarkers for the diagnosis and treatment of CRC.

S100A10 as a novel biomarker in colorectal cancer

Recent data support the role of S100A10 in tumorigenesis. In this study, we evaluated the value of S100A10 positivity as a possible biomarker in colorectal cancer. We evaluated S100A10 positivity by immunohistochemistry in a large population of colorectal cancer patients (n = 882). The relationships between S100A10 positivity and clinicopathological features and clinical outcome were analyzed. There were 36 % (319/882) tumors positive for S100A10 in all colorectal cancer samples. In contrast, normal colorectal epithelium was negative for S100A10 among all 562 specimens of adjacent normal mucosa. S100A10 positivity was correlated with poor differentiation (p = 0.0012) and disease stage (p = 0.003). S100A10 positivity was significantly correlated with shortened specific [log-rank p < 0.001; multivariate hazard ratio (HR), 1.49; 95% confidence interval (CI), 1.09-2.04] and overall survival (log-rank p = 0.0012; multivariate HR, 1.34; 95% CI, 1.06-1.73). Knockdown of S100A10 by siRNA significantly reduced the proliferation, migration, and invasion capacity of colorectal cancer cell lines. Our results suggest a role for S100A10 as a prognostic marker and potential therapeutic target in colorectal cancer.

Proteomic and cytokine plasma biomarkers for predicting progression from colorectal adenoma to carcinoma in human patients

In the present study, we screened proteomic and cytokine biomarkers between patients with adenomatous polyps and colorectal cancer (CRC) in order to improve our understanding of the molecular mechanisms behind turmorigenesis and tumor progression in CRC. To this end, we performed comparative proteomic analysis of plasma proteins using a combination of 2DE and MS as well as profiled differentially regulated cytokines and chemokines by multiplex bead analysis. Proteomic analysis identified 11 upregulated and 13 downregulated plasma proteins showing significantly different regulation patterns with diagnostic potential for predicting progression from adenoma to carcinoma. Some of these proteins have not previously been implicated in CRC, including upregulated leucine-rich α-2-glycoprotein, hemoglobin subunit β, Ig α-2 chain C region, and complement factor B as well as downregulated afamin, zinc-α-2-glycoprotein, vitronectin, and α-1-antichymotrypsin. In addition, plasma levels of three cytokines/chemokines, including interleukin-8, interferon gamma-induced protein 10, and tumor necrosis factor α, were remarkably elevated in patients with CRC compared to those with adenomatous polyps. Although further clinical validation is required, these proteins and cytokines can be established as novel biomarkers for CRC and/or its progression from colon adenoma.

MicroRNA-21 as a potential colon and rectal cancer biomarker

Colorectal cancer (CRC) is one of the most common malignant diseases worldwide and the prognosis is still poor although much progress has been achieved in recent years. In order to reduce CRC-related deaths, many studies are aimed at identifying novel screening- and prognosis-related biomarkers. MicroRNAs (miRNAs) are a class of 18-27-nucleotide single-stranded RNA molecules that regulate gene expression at the post-transcriptional level. It has been demonstrated that miRNAs regulate a variety of physiological functions, including development, cell differentiation, proliferation, and apoptosis. They play important roles in various physiologic and developmental processes and in the initiation and progression of various human cancers. It has been shown that miRNAs can critically regulate tumor cell gene expression, and evidence suggests that they may function as both oncogenes and tumor suppressor genes. In CRC, miRNAs-21 is one of the most important miRNAs and is rapidly emerging as a novel biomarker in CRC, with good potential as a diagnostic and therapeutic target. In this review, we summarize the latest research findings of the clinicopathological relevance of miRNAs-21 in CRC initiation, development, and progress, highlighting its potential diagnostic, prognostic, and therapeutic application, as well as discussing future prospects.

Identification of MXRA5 as a novel biomarker in colorectal cancer

In our previous study, significantly high expression levels of matrix-remodeling associated 5 (MXRA5) were identified in fresh-cultured colorectal cancer (CRC) tissues compared with their normal adjacent mucosa by differential secretome analysis. Whether MXRA5 is a potential serum biomarker of CRC has not been evaluated. The aim of this study was to investigate the association between MXRA5 expression and clinicopathological characteristics of CRC patients. The MXRA5 expression levels were determined by quantitative real-time PCR (qRT-PCR) and immunohistochemistry (IHC) in 20 colorectal adenoma tissues, 156 CRC tissues and their corresponding adjacent normal mucosa. Relative quantity (RQ) value and immunoreactive score (IRS) were used for quantitative assessment. The staining for MXRA5 protein was mainly located in the cytoplasm of CRC cells. All CRC tissues were positively stained, with a higher expression rate (IRS>4) of 67% (105/156), and a lower expression rate (IRS≤4) of 33% (51/156). Meanwhile, their corresponding normal tissues exhibited little positive staining; the higher expression rate was 0% (0/156) and the lower expression rate was 25% (16/156). Additionally, more than half of the adenoma tissues were positively stained; the higher expression rate was 15% (3/20) and the lower expression rate was 50% (10/20). The MXRA5 protein positive staining rates were significantly correlated with the lesion sites (colon vs. rectum, 76 vs. 59%), TNM staging (I+II vs. III+IV, 56 vs. 73%) and metastasis (present vs. absent; 76 vs. 61%) with the most high positive staining rate observable in omental metastasis (82%). However, MXRA5 mRNA expression levels showed no significant differences between CRC tissues and their corresponding normal tissues, and no significant correlation between IRS and corresponding RQ value was observed. In this study, we present the first evaluation of MXRA5 protein expression in CRC tissue. Our results revealed that MXRA5 protein is aberrantly expressed in CRC tissues, and has potential value in early detection of CRC and prediction of omental metastasis.

Serum microRNA-29a is a promising novel marker for early detection of colorectal liver metastasis

Background: Colorectal cancer (CRC) metastasis occurs in various organs, most frequently in liver. Serological examination including tumor and biochemical markers for liver function evaluation is routinely performed, though its accuracy is not high. MicroRNAs (miRNAs) have been implicated in a variety of human diseases including cancer, and have many characteristics of an ideal biomarker most notably their inherent stability and resilience. Recently, several studies have indicated that circulating miRNAs hold much potential as novel noninvasive biomarkers for cancer and other disease processes. The objective of this study was to investigate the potential of serum miRNAs as novel biomarkers for CRC with liver metastasis. Methods: This study was divided into three phases: (I) 3 candidate serum miRNAs were detected by using real-time RT-PCR, corresponding 38 CRC patients with liver metastasis and 36 CRC patients without metastasis. (II) Marker validation by real-time RT-PCR on a similar cohort of age- and sex-matched CRC patients without ( n = 20) and with liver metastasis ( n = 20). (III) We examined the correlation between the expressions of candidate serum miRNAs with clinical parameters of CRC patients. Results: Serum miR-29a was significantly higher in colorectal liver metastasis (CRLM) patients than in CRC patients. This marker yielded a receiver operating characteristic curve area of 80.3%. At a cutoff value of 0.155, the sensitivity was 75% and the specificity was 75% in discriminating metastatic from non-metastatic patients. In addition, increased levels of miR-29a expression were also observed in colorectal tumors from CRLM patients compared with CRC patients. No significant difference was observed in the levels of serum miR-92a between metastatic and non-metastatic patients. Conclusions: These findings suggest that serum miR-29a has strong potential as a novel noninvasive biomarker for early detection of CRC with liver metastasis.

MicroRNA Signatures: Novel Biomarker for Colorectal Cancer?

Aberrant microRNA (miRNA) expression might be of potential use as diagnostic and prognostic biomarker for cancers. We reviewed studies published until March 2011 which assessed expression of miRNAs in colorectal cancer (CRC)/adenoma tissue and normal colorectal mucosa and in plasma of CRC/adenoma patients and healthy controls. Overall, 20 studies that investigated miRNA expression in tissue and 3 studies that investigated miRNA levels in plasma were included. A total of 160 miRNAs were found to be dysregulated in CRC. MiR-20a and miR-31 were found to be significantly upregulated in more than one study, and miR-143 and miR-145 were found to be significantly downregulated in CRC tissue in six or more studies. MiR-92a was significantly upregulated in CRC patients in two of the plasma-based studies and in CRC tissue in one of the tissue-based studies. Our results provide timely and relevant information for miRNAs as potential diagnostic biomarkers for CRC. The expression of miRNAs in plasma may be indicative of presence of CRC. Larger diagnostic studies are needed to evaluate potential use of miRNA expression in early detection and diagnosis of CRC.

Importins Are Critical for Colorectal Cancer (CRC) Growth and Are Novel Biomarkers of CRC. Underlying Mechanisms Include: Increased Nuclear Transport of P-CREB and P-STAT3, VEGF Gene Promoter Activation and Aberrant VEGF Expression

Importins Are Critical for Colorectal Cancer (CRC) Growth and Are Novel Biomarkers of CRC. Underlying Mechanisms Include: Increased Nuclear Transport of P-CREB and P-STAT3, VEGF Gene Promoter Activation and Aberrant VEGF Expression

Sperm-Associated Antigen 9 Is a Novel Biomarker for Colorectal Cancer and Is Involved in Tumor Growth and Tumorigenicity

Colorectal cancer (CRC) is the second most common tumor in developed countries. The present study was undertaken to determine the expression of the sperm-associated antigen 9 gene (SPAG9) as a possible biomarker in CRC, to investigate its correlation with humoral immune response and different stages and grades in CRC patients, and to explore its possible role in colon tumorigenesis in vitro and in an in vivo mouse model. SPAG9 expression was determined by RT-PCR, in situ RNA hybridization, and immunohistochemistry. Humoral response against SPAG9 was detected by enzyme-linked immunosorbent assay and Western blotting. SPAG9 gene silencing was performed using plasmid-based small interfering RNA to study various malignant properties of colon cancer cells in vitro and in vivo. The majority of CRC patients showed SPAG9 expression and generated humoral response. There was a close relationship between SPAG9 protein expression and humoral immune response in the majority of early-stage CRC patients, indicating that anti-SPAG9 antibodies could be a novel serum biomarker for early diagnosis. The down-regulation of SPAG9 (mediated by small interfering RNA) inhibited malignant properties in in vitro and significantly suppressed tumor growth in vivo. These findings collectively suggest that SPAG9 may have a role in tumor development and early spread and thus could serve as a novel target for early detection and for cancer immunotherapy.

Clinical Validation of Colorectal Cancer Biomarkers Identified from Bioinformatics Analysis of Public Expression Data

Identification of novel biomarkers of cancer is important for improved diagnosis, prognosis, and therapeutic intervention. This study aimed to identify marker genes of colorectal cancer (CRC) by combining bioinformatics analysis of gene expression data and validation experiments using patient samples and to examine the potential connection between validated markers and the established oncogenes such as c-Myc and K-ras. Publicly available data from GenBank and Oncomine were meta-analyzed leading to 34 candidate marker genes of CRC. Multiple case-matched normal and tumor tissues were examined by RT-PCR for differential expression, and 9 genes were validated as CRC biomarkers. Statistical analyses for correlation with major clinical parameters were carried out, and RNA interference was used to examine connection with major oncogenes. We show with high confidence that 9 (ECT2, ETV4, DDX21, RAN, S100A11, RPS4X, HSPD1, CKS2, and C9orf140) of the 34 candidate genes are expressed at significantly elevated levels in CRC tissues compared to normal tissues. Furthermore, high-level expression of RPS4X was associated with nonmucinous cancer cell type and that of ECT2 with lack of lymphatic invasion while upregulation of CKS2 was correlated with early tumor stage and lack of family history of CRC. We also demonstrate that RPS4X and DDX21 are regulatory targets of c-Myc and ETV4 is downstream to K-ras signaling. We have identified multiple novel biomarkers of CRC. Further analyses of their function and connection to signaling pathways may reveal potential value of these biomarkers in diagnosis, prognosis, and treatment of CRC.

MicroRNAs are novel biomarkers of colorectal cancer

Recent studies have identified unique small ribonucleic acids called microRNAs (miRNAs) in colonic tumour tissue and blood that may accurately diagnose the presence of colorectal cancer and help predict disease recurrence. This review explores the potential role of these biomarkers. A literature search identified studies describing miRNAs in colorectal cancers. The outcomes of interest included diagnosis, progression and recurrence of disease, and future therapy. Overexpression and silencing of specific miRNAs are associated with the development and progression of colorectal cancer. Such a role in oncogenesis suggest that miRNAs may be important targets for gene therapies. Differential expression of specific miRNAs in tissues and blood offers the prospect of their use in early detection and screening for colorectal cancer. MiRNAs are implicated in metastasis and cytotoxic drug resistance. Their manipulation has potential in both prevention of recurrence and palliation. The miRNAs expression profile in tissue and blood has potential for their use in the detection, screening and surveillance of colorectal cancer. Furthermore, miRNAs may be targeted by gene therapy to treat colorectal cancer.

Up-regulation and clinical relevance of novel helicase homologue DHX32 in colorectal cancer

BackgroundThis study aimed to find novel biomarkers for colorectal cancer.MethodsFluorescent mRNA differential display PCR (DD-PCR) was used to screen the genes differentially expressed in colorectal cancer tissues and their adjacent tissues. The differentially expressed genes were confirmed by real-time PCR and then their clinical relevance (such as association with tumor location and lymph gland metastasis) was further investigated.ResultsWe identified by DD-PCR a novel RNA helicase, DHX32, which showed higher expression in colorectal cancer tissues than their adjacent tissues, and this result was confirmed by real time RT-PCR. In addition, we found that the level of DHX32 gene expression in colorectal cancer was significantly associated with cancer location, lymph gland metastasis, cancer nodal status, differentiation grade, and Dukes, stage.ConclusionDHX32 may play an important role in the development of colorectal cancer and could serve as a novel biomarker for colorectal cancer after additional investigation.