Prognosis Of Colorectal Carcinoma Research Articles

The role and gene expression profile of SOCS3 in colorectal carcinoma.

SOCS3 has been postulated to play a role in the occurrence and progression of malignancies. However, the relationship of SOCS3 with colorectal carcinoma remains poorly understood. The purpose of the study was to explore the role of SOCS3 in colorectal carcinoma and its underlying mechanisms. Protein and mRNA expression of SOCS3 in colorectal carcinoma and normal colorectal mucosa was detected using immunohistochemistry and real-time quantitative PCR. SOCS3 expression was significantly lower in colorectal carcinoma tissue than in normal colorectal mucosa, and was negatively correlated with tumor invasion depth, lymph node metastasis, differentiation degree, and TNM stage. A stably transfected colorectal carcinoma cell line (8348SOCS3) with high expression of SOCS3 was established. The effects of SOCS3 overexpression on the growth, proliferation, invasion and tumor formation of colorectal carcinoma cells were examined by CCK-8 assay, transwell method and tumorigenicity assays in nude mice. Then we found SOCS3 overexpression significantly decreased proliferation and invasion capability of 8348 cells in vitro and in vivo. Furthermore, the effect of SOCS3 overexpression on the gene expression profile of colorectal carcinoma cells was analyzed using human genome arrays. The results revealed 369 genes that were differentially expressed in 8348SOCS3 cells. 193 genes was significantly increased and 176 genes was significantly decreased. Bioinformatics analysis demonstrated that high SOCS3 expression affected multiple signaling pathways in colorectal carcinoma including TGF-β/Smads, NF-κB, and HIF-MAPK pathways. Especially for the TGF-β/Smads pathways, high SOCS3 expression could inhibit TGF-β1 expression and activate Smad4 expression. These data suggested that low expression of SOCS3 was associated with the occurrence and progression of colorectal carcinoma. SOCS3 protein may be a useful indicator for malignancy and prognosis of colorectal carcinoma and also a new target for gene therapy.

Elevated serum level of lncRNA-HIF1A-AS1 as a novel diagnostic predictor for worse prognosis in colorectal carcinoma.

To evaluate the diagnostic efficacy and prognostic value of serum long non-coding RNA (lncRNA) HIF 1alpha-antisense RNA 1 (HIF1A-AS1) in patients with colorectal carcinoma (CRC). We obtained serum samples from 151 CRC patients and 160 health controls. Serum level of HIF1A-AS1 was detected via real-time PCR (RT-PCR). Receiver operating characteristics (ROC) curve analysis was conducted to determine the diagnostic value of HIF1A-AS1. Then HIF1A-AS1 in CRC was divided into high- and low-expression groups, and the associations of the HIF1A-AS1 serum level with clinicopathological features and prognosis were analyzed. Serum level of HIF1A-AS1 was significantly increased from CRC patients as compared to those of health controls (P< 0.05). ROC curve analysis revealed a relative high diagnostic performance of HIF1A-AS1 to distinguish CRC from health controls, with the area under the curves (AUC) of 0.960 (95% CI: 0.940 ∼ 0.980; P< 0.001). Kaplan-Meier analysis showed that differentiation degree, tumor size, TNM stage, T stage, N stage, M stage and serum level of HIF1A-AS1 were all linked to CRC prognosis (All P< 0.05). Compared to CRC patients with low HIF1A-AS1 expression, high expression of patients were associated with a shorter 5-year-survival rate (P< 0.001). Multivariate Cox regression analysis revealed that lower differentiation degree, tumor > 5cm and higher expression of HIF1A-AS1 were independent risk factors affecting the survival rate of patients with CRC (P< 0.05). Our results illustrated that elevated serum HIF1AAS1 could be clinically functioned as a potential biomarker for CRC diagnoses and prognosis.

LncRNA FAM83H‑AS1 is associated with the prognosis of colorectal carcinoma and promotes cell proliferation by targeting the Notch signaling pathway

The long non-coding RNA, FAM83H antisense RNA 1 (head to head) (FAM83H-AS1), has exhibited a functional role as an oncogene in a number of different types of cancer. The aim of the present study was to reveal the dysregulation of FAM83H-AS1 in colorectal carcinoma (CRC) samples and elucidate its underlying associations with the Notch signaling pathway. The expression profiles of FAM83H-AS1 and two Notch signaling-associated molecules, Notch1 and Hes family basic-helix-loop-helix transcription factor 1 (Hes1), were measured by reverse transcription-polymerase chain reaction and western blot analysis. The Pearson χ2 test was employed to evaluate the associations between FAM83H-AS1 expression and clinical features. A statistically significant positive association between the expression levels of FAM83H-AS1 and those of Notch1 or Hes1 in CRC tissues was analyzed by Spearman's correlation analysis. The Kaplan-Meier method was used to compare the overall survival curves between the highly-expressed and low-expressed FAM83H-AS1 groups via a log-rank test. Specific small hairpin RNA was transfected to silence endogenous FAM83H-AS1. MTT and colony formation assays were performed to measure the growth-inhibition effect of silenced FAM83H-AS1. The levels of FAM83H-AS1, Notch1 and Hes1 were significantly increased in CRC samples and cell lines. Cell proliferation was markedly inhibited when FAM83H-AS1 was knocked down and this effect mediated by FAM83H-AS1 could be reversed by Notch1 regulators. Thus, downregulated FAM83H-AS1 exhibited an anti-proliferative role in CRC by repressing the Notch signaling pathway.

Liver Imaging for Colorectal Cancer Metastases

Purpose of Review The prognosis of colorectal carcinoma with liver metastases (CRLM) is driven largely by the resectability of hepatic metastatic disease. An increasing number of systemic and hepatic-directed therapies are being applied in the neoadjuvant setting to downstage patients for surgical eligibility. The goal of this review is to describe the use of imaging techniques in the management of CRLM with an emphasis on staging, pretreatment planning, and response assessment.

Detection and clinical significance of circulating tumor cells in patients with colorectal carcinoma

Objective: To detect circulating tumor cells (CTC) in patients with colorectal carcinoma and to evaluate the relationship among CTC, clinic-pathological characteristics and prognosis of colorectal carcinoma. Methods: Peripheral blood samples were obtained from 109 patients with colorectal carcinoma in Department of General Surgery, Beijing Friendship Hospital, Capital Medical University from April 2014 to October 2016. There were 60 male and 49 female patients, aging from 33 to 86 years with a mean age of (65±10) years.CTC were detected using density-gradient centrifugation and immunofluorescence staining. χ(2) test, Fisher exact test and rank-sum test were used to analyze the relation between positive rate of CTC and clinical characteristic, respectively. The correlation analysis of CTC and common tumor markers was detected by χ(2) test and Spearman test. The overall survival of patients was analyzed by Kaplan-Meier curve and Cox proportional hazard model. Results: CTC were found in 71 of the 109 patients with colorectal carcinoma. The presence of CTC was significantly correlated with N stage (Z=4.422, P=0.035) and M stage (χ(2)=4.424, P=0.049). However, CTC was not significantly correlated with age, sex, tumor location, tumor size, differentiation, T stage, Ki-67 and TNM stage (P>0.05). Meanwhile, there was significant correlation between CTC and carcino-embryonic antigen (CEA) (χ(2)=4.897, P=0.027; r=0.212, P=0.027) indicated by χ(2) test and Spearman correlations analysis. The positive rate of CTC was higher than that of CEA (χ(2)=15.45, P=0.000). Survival analysis suggested that positive CTC was poor for overall survival in colorectal cancer with adjusted HR as 3.023(95%CI: 1.330 to 6.872, P=0.008). Conclusions: CTC is helpful to early diagnosis tumor recurrence and metastasis. Hence, combined multiple tumor markers, including the CTC as common indicators of tumor diagnosis, relapse and metastasis could effectively improve the accuracy of diagnosis.

Risk score based on ten lncRNA-mRNA expression predicts the survival of stage II-III colorectal carcinoma.

The prognosis of colorectal carcinoma (CRC) is unstable in the stage II-III patients. Patients with early stage II CRC have a relative poor prognosis while other stage III CRC patients have a better prognosis. In our work, by utilizing the expression of lncRNAs and mRNAs measured by microarray (GSE39582), we constructed a risk score staging system with Cox multivariate regression model to predict the outcome of grade II-III CRC patients. Ten genes including two lncRNAs and eight mRNAs were used to estimate the survival of stage II-III CRC patients. The patients with high risk scores have poorer survival rate those with low risk scores, significantly. These results were further validated in another three independent datasets (GSE37892, GSE33113, and GSE17536). The relationship between clinical information and were evaluated, and the risk score is independent from the other clinical information and performs better in evaluating the survival of stage II-III CRC patients. Moreover, the correlation between chemotherapy was also evaluated, and we found that both patients with or without chemotherapy have a poor survival in high risk group. Gene Set Enrichment Analysis were used to find the difference between high-risk and low-risk groups, and pathways including cell adhesion and focal adhesion were significantly enriched, suggesting that the risk score reflects the status of cell-cell physical interaction. In summary, we constructed a risk staging model for grade II-III CRC, which is independent from and performs better than clinical information.

Increased expression of BPI fold-containing family A member 1 is associated with metastasis and poor prognosis in human colorectal carcinoma.

Bactericidal or permeability-increasing protein fold-containing family A member 1 (BPIFA1) has been demonstrated to be involved in inflammatory responses in the upper airway and the progression of non-small cell lung cancer. However, the expression levels of BPIFA1 and its clinical prognostic significance in colorectal carcinoma (CRC) has not yet been elucidated. Reverse transcription-polymerase chain reaction and immunohistochemistry were used to analyze the expression levels of BPIFA1 in CRC and normal mucosal tissues. The associations between BPIFA1 expression levels and clinicopathological characteristics, and its predictive value for prognosis in CRC, were statistically evaluated as appropriate. The expression levels of BPIFA1 were revealed to be upregulated at the transcriptional and translational levels in CRC tissues, compared with in normal mucosal tissues. A high expression level of BPIFA1 is significantly associated with invasion depth (P=0.040), lymph node metastasis (P=0.035) and distant metastasis (P=0.010). Furthermore, Kaplan-Meier analysis indicated that BIPFA1 overexpression is associated with short survival time, and the Cox proportional hazards model of risk analysis indicated that BPIFA1 is an independent prognostic factor for patients with CRC. The results of the present study suggested that BPIFA1 expression is upregulated in CRC tissues, and that an increased expression level of BPIFA1 is associated with tumor invasion, metastasis and poor prognosis, indicating that BPIFA1 may be a potential clinical prognostic predictor and therapeutic target for patients with CRC.

Expression of family with sequence similarity 172 member A and nucleotide-binding protein 1 is associated with the poor prognosis of colorectal carcinoma.

In our previous studies, a functionally unknown gene, family with sequence similarity 172, member A (FAM172A), was identified. High levels of FAM172A suppressed the cell cycle process, arresting HepG2 cells in G1/S and inhibiting cell proliferation. The present study aimed to confirm the expression levels of FAM172A and nucleotide-binding protein 1 (NUBP1) in colorectal cancer (CRC) tissues and normal colorectal tissues. The impact of FAM172A and NUBP1 on the prognosis of patients with CRC was also analyzed. Immunohistochemical staining for FAM172A and NUBP1 was performed on 180 cancerous tissues and 60 normal paraffin-embedded tissues from patients with CRC. In total, 85 and 83% of 180 patients revealed positive expression of FAM172A and NUBP1, respectively. FAM172A expression level was associated with Tumor-Node-Metastasis (TNM) staging (P<0.001), the levels of serum carcinoembryonic antigen (CEA; P=0.023) and carbohydrate antigen 19–9 (CA19-9; P=0.016), lymph node involvement (P=0.004), tissue type (P=0.016), Dukes' staging (P<0.001) and NUBP1 (P=0.026). Furthermore, the expression level of NUBP1 was also markedly associated with the levels of serum CEA (P=0.006) and CA19-9 (P=0.001), TNM staging (P<0.001), lymph node involvement (P=0.005), histological typing (P=0.024) and Dukes' stage (P<0.001). Results of the univariate analysis demonstrated that there was a negative correlation between the expression level of FAM172A and overall survival (OS) and relapse-free survival (RFS) (P=0.013 and P=0.012, respectively), and there was also a negative correlation between NUBP1 expression level and OS and RFS (P<0.001 and P<0.001, respectively). With regards to OS and RFS, multivariate analysis revealed that expression levels of FAM172A and NUBP1 and tumor stage may be independent prognostic factors Thus, the present study suggested that FAM172A and NUBP1 may be prognostic makers for CRC.

Abstract 4723: Improving prognosis or promoting progression? Double sword of S100A8 in colorectal cancer

Abstract Tumor tissues are heterogeneous, including cancer cells, cancer stem cells and immune cells. The immune states are involved in malignancy and prognosis in colorectal carcinomas (CRC). S100A8, which represented granulocytes and monocytes, was identified as a favorable indicator for survival by previous work. Cancer stem cells (CSCs) and epithelial-mesenchymal transition (EMT) play critical roles on tumor cell malignancy and metastasis. The immune states, CSC and EMT should be regarded as integrity, which is important for clinical efforts and the mechanism investigation. For better understanding the associations among immune cells, CSC and EMT in CRC, S100A8 as along with CSC (Lgr5, Nanog, ALDH1 and CD44v6) and EMT (E-cadherin and Snail) markers were detected in the tumor invasive front (TF) and tumor center (TC) respectively by immunohistochemistry in 419 CRCs and survival analysis was done. Furthermore, functions of S100A8 in EMT were verified in vitro. S100A8+TF count was positively correlated with Lgr5TC (r=0.162, p=0.004) and cytoplasmic SnailTF expression (r=0.116, p=0.043), while it was not correlated with other stem cell markers wherever they were located, and not associated with E-cadherinTC or E-cadherinTF, or cytoplasmic snailTC or nucleic SnailTC and SnailTF expression (p&amp;gt;0.005). After combining S100A8+TF and Lgr5TC, three groups were got. It showed S100A8+TFhigh Lgr5TChigh had the best 5-year survival, while S100A8+TFlow Lgr5TClow had the worst 5-year survival. After joining with cytoplasmic snailTF into S100A8+TF and Lgr5TC, six groups were got. Finally, three groups with similar outcome were merged as A, B, and C, called SLS (S100A8, Lgr5 and snail). Kaplan-Meier survival curve analysis showed group C, with S100A8lowLgr5lowsnailhigh had the worst 5-year survival. SLS was also an independent predictor for 5-year survival. S100A8 was not only stained in granulocytes and monocytes but also in tumor cells. Then S100A8 was overexpressed by lentiviral vector in DLD1 and sw620 CRC cell lines to further investigate its intracellular functions. On the contrary, overexpression of S100A8 in tumor cells not only facilitated tumor cell proliferation, migration and invasion but also converted cells to EMT phenotype. Western blot showed E-cadherin was down-regulated and Vimentin, MMP9 and snail were up-regulated in S100A8 overexpressed cells. The transcription factor USF2 was verified to interact with S100A8 by luciferase reporter gene. Western blot showed overexpression of USF2 increased S100A8 expression, while overexpression of S100A8 had no effect on USF2. In conclusion, S100A8 from immune cells or tumor cells has opposing effects on cancer progression. S100A8 in immune cells predicts good prognosis. However, S100A8 in tumor cells, which could be regulated by USF2, promotes proliferation, migration and invasion, along with EMT transformation. Citation Format: Si Li, Fangying Xu, Lili Wang, Hui Li, Maode Lai. Improving prognosis or promoting progression? Double sword of S100A8 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 4723. doi:10.1158/1538-7445.AM2017-4723

Reduced RCE1 expression predicts poor prognosis of colorectal carcinoma

BackgroundAs an end-proteolytic enzyme that cleaves the last three residues of proteins with a terminal CAAX, Ras-converting enzyme 1 (RCE1) has an essential role in multiple signaling pathways and take part in the process of differentiation, proliferation and carcinogenesis. The aim of the study is to investigate expression pattern of RCE1 and its prognosis in colorectal carcinoma (CRC).MethodsThe expression of RCE1 and phospho-MAPK family members was confirmed by immunohistochemical staining of CRC tissues. miR-RCE1 lentiviral vectors were transduced into HCT116 and SW489 cells. Reverse transcription PCR (RT-PCR) and western blot were conducted to measure the transfection efficiency. Transwell assays were used to detect the invasiveness of CRC cells.ResultsIn the present study, we assessed RCE1 expression in 244 CRC specimens and matching adjacent, non-tumorous tissues by immunohistochemistry (IHC). Compared with the matched adjacent non-tumor tissue samples, the RCE1 reduced in the tumor tissue samples (p < 0.001). RCE1 expression was significantly decreased in 106 of 244 (43.4%) CRC cases. In univariate and multivariate analyses, Decreasing expression of RCE1 independently predicts poor prognosis for patients in both overall survival and disease-free survival. Further study indicated that RCE1 influenced tumor invasion through the p38 pathway. Knockdown of RCE1 reduced phosphorylation and significantly increased the invasive capacity of CRC cells.ConclusionTaken together, the outcomes of this study indicate that RCE1 acts as a tumor suppressor in CRC, as its reduced expression may increase CRC cell invasion and independently predict an unsatisfactory prognosis in CRC patients.

Intraepithelial CD94+ tumor-infiltrating lymphocytes in a context of HLA-E overexpression as a new immune checkpoint to predict prognosis in colorectal carcinomas.

e14592 Background: A better understanding of the immune-modulating interactions between tumor cells and immune cells underlying the balance between immune control and immune resistance in colorectal cancer (CRC) is crucial for the design of immunotherapies. We have previously demonstrated that overexpression of the human MHC class Ib molecule - HLA-E/β2 microglobulin - by tumor cells in CRC was associated with an unfavorable prognosis, suggesting its involvement in immune escape. However, the specific receptor of HLA-E/β2m - CD94/NKG2A, inhibitory or CD94/NKG2C, activating - expressed by tumor-infiltrating lymphocytes (TIL), as well as the influence of the microsatellite status in HLA-E/β2m overexpression, remain unknown. Methods: We investigated in the primary tumor of 245 CRC patients 1) the association of HLA-E/β2m overexpression and the density of CD94+ intraepithelial TIL (IEL-TIL) with the microsatellite status, 2) the nature of CD94+ TIL and the receptor expressed - CD94/NKG2A or CD94/NKG2C - and 3) the prognostic influence of CD94+ IEL-TIL. Results: HLA-E/β2m was preferentially overexpressed in MSI compared with MSS CRC (44,6 % vs 18,4 % respectively, p = 0.0001), and significantly associated with a high density of CD94+ IEL-TIL in MSI (0,9% in HLA-E/β2m+ vs 0,2% in HLA-E/β2m– CRC, p = 0,001), and in MSS CRC (0,38% vs 0,15%, p &lt; 0,0001). These CD94+ TIL mostly corresponded to CD8+ αβ T cells preferentially expressing the inhibitory NKG2A chain. Finally, a high density of CD94+ IEL-TIL was independently associated with a worse OS (p = 0.03). Conclusions: These results strongly suggest that HLA-E/β2m - CD94/NKG2A interactions, preferentially up-regulated in MSI CRC, represent a promising inhibitory immune checkpoint. From a clinical point of view, this inhibitory immune checkpoint could be blocked by the new anti-NKG2A monoclonal antibody.

Evaluation of the correlation of vasculogenic mimicry, ALDH1, KAI1 and microvessel density in the prediction of metastasis and prognosis in colorectal carcinoma

BackgroundMetastasis and recurrence are the most common reasons for treatment failure of colorectal carcinoma (CRC). Vasculogenic mimicry (VM, blood supply formation often seen in highly aggressive tumors), Aldehyde dehydrogenase 1 (ALDH1, a biomarker of cancer stem cells), KAI1 (a suppressor gene of tumor metastasis) are all valuable factors for metastasis and prognosis in diverse human cancers. However, the correlation of VM, ALDH1, KAI1 and microvessel density (MVD) in CRC is unclear. In this study, we analyzed the correlations among VM, ALDH1, KAI1 and MVD, as well as their respective correlations with clinicopathological parameters and survival in CRC.MethodsThe level of VM, ALDH1, KAI1 and MVD in 204 whole tissue samples of CRC were examined by immunhistochemistry. Clinical data was also collected.ResultsLevels of VM, ALDH1 and MVD were significantly higher, and levels of KAI1 significantly lower, in CRC tissues than in normal colorectal tissues. Levels of VM, ALDH1 and MVD were positively associated with invasion of depth, lymph node metastasis (LNM), distant metastasis and tumor-node-metastasis (TNM) stages, and negatively with patients’ overall survival (OS). Levels of KAI1 was negatively correlated with invasion of depth, LNM, distant metastasis and TNM stages, and the KAI1 positive expression subgroup had significantly longer OS than did the KAI1- subgroup. In multivariate analysis, high levels of VM, ALDH1 and KAI1, as well as TNM stages were independently correlated with lower OS in patients with CRC.ConclusionsVM, MVD and the expression of ALDH1 and KAI1 may represent promising metastatic and prognostic biomarkers, as well as potential therapeutic targets for CRC.

Expression of Cyclin E, proliferation cell nuclear antigen and matrix metalloproteinase-9 in colorectal carcinoma cells and the significance

Objective To explore the effect of the expression of cell cycle regulators on the development and prognosis of colorectal carcinoma. Methods The surgicall resected tissues from 128 cases of colorectal cancer were collected in our hospital from June 2012-December 2015 December. The expression levels of Cyclin E, proliferation cell nuclear antigen (Ki-67) and matrix metalloproteinase (MMP)-9 were detected by immunohistochemieal staining in the large intestine mucosa from 128 cases of carcinoma and 24 samples of normal mucosa.χ2 test, single factor analysis of variance, Kaplan-Meier survival rate curve analysis and Log-rank test were applied.P<0.05 was considered to be statistically significant difference. Results The positive rate of Cyclin E, Ki-67 and MMP-9 was significalntly higher in colorectal carcinoma than in normal mucosa (χ2=5.203, P=0.013). The expression of Cyclin E in Duke’s stage A was markedly higher than in Duke’s stages B, C and D (χ2=10.331, P=0.001), and it was higher in cases with negative lymph node metastasis than in those with positive lymph node metastasis (χ2=6.732, P=0.005). The expression of Cyclin E was not correlated with the prognosis of the cases with lymph node metastasis. The down-regulated expression or deficiency of MMP-9 was related to advanced Duke’s stages (χ2=7.201, P=0.003) and lymph node metastasis (χ2=12.072, P=0.000). The Ki-67 expression was not associated with the carcinoma localization, advanced Duke’s stages and prognosis. There were 44 deaths from 128 patients, accounting for 39.29%, with 24, 14 and 36 cases positive for, and 20, 30 and 8 cases negative for Cyclin E, Ki-67 and MMP-9 expression, respectively. The 5-year survival rate in Cyclin E positive and negative expression groups was 63.64% and 56.52% respectively, with no significant difference (χ2=0.958, P=0.273). The 5-year survival rate in MMP-9 positive expression group was 73.33%, significantly higher than that in MMP-9 negative group (46.15%, χ2=3.143, P=0.035). The 5-year surfival rate in Ki-67 positive group (61.70%) and negative group (55.56%) showed no significant difference (χ2=0.827, P=0.419). The expression of Cyclin E was positively correlated with Ki-67 (r=0.391, P=0.019), MMP-9 and Ki-67 showed no significant correlation (r= -0.109, P=0.387), and MMP-9 and Cyclin E expression had no significant correlation (r=-0.168, P=0.183). Conclusion The overexpression of Cyclin E and decline or deficiency of MMP-9 expression might accelerate the progression of the cell cycle and promote the carcinogenesis of colorectal carcinoma. The overexpression of Cyclin E might be a relatively early event in initiation of colorectal carcinoma. The decline or deficiency of MMP-9 expression might promote the development and metastasis of colorectal carcinoma, and it could be an indicator for unfavorable prognosis. Key words: Colorectal carcinoma; Cyclin E; Matrix metalloproteinase-9; Cell cycle; Prognosis

S100A8+ stroma cells predict a good prognosis and inhibit aggressiveness in colorectal carcinoma

ABSTRACTGene microarray and bioinformatic analysis showed that S100A8 was more abundant in the stroma surrounding tumor buddings (TBs) than in the stroma surrounding primary tumor cells in colorectal carcinomas. Here, S100A8+ cells in 419 colorectal carcinoma samples were stained by immunohistochemistry and counted using Image-pro plus 6.0. TBs were also counted and biomarkers associated with the epithelial–mesenchymal transition and apoptosis were assessed by immunohistochemistry. We evaluated the association between S100A8+ cells and clinico-pathological variables as well as survival. Migration and invasion as well as biomarkers of the epithelial–mesenchymal transition and apoptosis were tested in CRC cells, treated with graded concentrations of recombinant human S100A8 protein.We found that the density of S100A8+ cells in the tumor invasive front (S100A8+TIF) clearly distinguished patients with 5-y survival from those who did not survive (p = 0.01). The S100A8+-associated tumor budding (SATB) index determined by the S100A8+TIF and TB was an independent predictor of overall survival (p = 0.001) other than the S100A8+TIF or TB alone. Migration and invasion properties of CRC cells were inhibited by recombinant human S100A8 treatment. The particular S100A8+ cells in the stroma were associated with important biomarkers of the epithelial–mesenchymal transition (E-cadherin and SNAIL) and apoptosis (BCL2).In conclusion, S100A8+ cells in the stroma predict a good prognosis in colorectal carcinoma. An index combining S100A8+ cells and TB independently predicts survival. Recombinant human S100A8 inhibited CRC cell migration and invasion, which was involved in epithelial–mesenchymal transition (E-cadherin and SNAIL) and apoptosis (BCL2).

Nuclear expression and/or reduced membranous expression of β-catenin correlate with poor prognosis in colorectal carcinoma: A meta-analysis.

The differential subcellular localizations of β-catenin (including membrane, cytoplasm, and nucleus) play different roles in the progression of colorectal cancer (CRC). However, the correlation between each subcellular localization of β-catenin and the prognosis of CRC patients remains undetermined. Systematic strategies were applied to search for eligible published studies in the PubMed, Embase, and Web of Science databases. The correlation between each subcellular localizations of β-catenin expression and patients' clinicopathological features or prognosis was analyzed. Finally, this meta-analysis, including 6238 cases from 34 studies, revealed that β-catenin overexpression in the nucleus (HR: 1.50[95% CI: 1.08-2.10]) or reduced expression of β-catenin in the membrane (HR: 1.33[95% CI: 1.15-1.54]) significantly correlated with lower 5-year overall survival (OS). Conversely, overexpression of β-catenin in the cytoplasm (HR: 1.00[95% CI: 0.85-1.18]) did not show significant association with 5-year OS. This study suggested that β-catenin overexpression in the nucleus or reduced expression in the membrane, but not its overexpression in cytoplasm, could serve as a valuable prognostic predictor for CRC. However, additional large and well-designed prospective studies are required to verify our results.

The relationship of A Proliferation-Inducing Ligand expression with the clinical pathology and prognosis in colorectal carcinoma

Objective: To investigate the protein expression of A Proliferation-Inducing Ligand (APRIL) in colorectal cancer (CRC) tissue, and its correlation with clinical pathology and prognosis. Methods: One hundred and twenty cases of CRC tissue and related clinical data were collected from the Department of Pathology of Affiliated Hospital of Nantong University. The specimens were made into tissue microarray. The protein expression of APRIL was detected by Envision immunohistochemistry. The relationship of the protein expression of APRIL with clinicopathological parameters and follow-up data were statistically analyzed. Results: APRIL protein expression positive rate of CRC tissue and cancer adjacent normal mucosal tissue were 83.33% (100/120) and 12.5% (15/120), with a statistical significance (P<0.001). The expression of APRIL was related to TNM staging (χ2=10.222, P=0.006) and the depth of tumor invasion (χ2=5.987, P=0.014). Kaplan-Meier survival curves showed that five years survival rate of high APRIL expression group was lower than that of low APRIL expression group. Univariate survival analysis showed that the expression of APRIL (P<0.001), the degree of tumor differentiation (P=0.033), TNM staging (P<0.001), depth of tumor invasion (P=0.011), lymph node metastasis (P<0.001) and serum CEA (P=0.003) time were associated with postoperative survival. Multivariate COX proportional hazard regression model analysis further revealed the expression of APRIL (P=0.001), the degree of tumor differentiation (P=0.005), TNM staging (P=0.001) and serum CEA (P=0.039) were the independent risk factors for prognosis. Conclusion: The expression of APRIL in CRC tumor tissue is closely related to TNM stage and depth of tumor invasion. The high expression of APRIL prompts bad prognosis. APRIL is one of the independent prognostic factors in patients with CRC, and could be a potential target for the therapy of CRC.

Patterns and prognostic relevance of PD-1 and PD-L1 expression in colorectal carcinoma

AbstractImmune checkpoint blockade targeting the programmed death-1 (PD-1) pathway has shown efficacy in several types of cancers including mismatch-repair-deficient colorectal carcinoma. In some tumor types, programmed death-ligand 1 (PD-L1) expression detected by immunohistochemistry has shown utility as a predictive marker for response to anti-PD-1 therapies. This utility, however, remains to be determined in colorectal carcinoma. In addition, although tumor-infiltrating lymphocytes have been associated with better prognosis in colorectal carcinoma, the prognostic value of PD-1 expression in these lymphocytes and its interaction with PD-L1 expression still await investigation. To address these questions, we performed a pilot study to evaluate the patterns of PD-L1 and PD-1 immunohistochemical expression on colorectal carcinoma cells and their tumor-infiltrating lymphocytes, respectively. Using tissue microarray, we found that 5% (19/394) of colorectal carcinomas exhibited high tumor PD-L1 expression, and 19% (76/392) had elevated numbers of PD-1-positive tumor-infiltrating lymphocytes. PD-L1 levels correlated with PD-1 levels (P<0.001), and mismatch-repair-deficient tumors had significantly higher rates of high PD-L1 and PD-1 expression when compared with mismatch-repair-proficient tumors (18% vs 2% and 50% vs 13%, respectively; P<0.001 for both). Staining intensity was also stronger for both markers in mismatch-repair-deficient tumors. Furthermore, we observed that among patients with mismatch-repair-deficient colorectal carcinoma, PD-1/PD-L1 expression stratified recurrence-free survival in an inter-dependent manner: an association between high PD-1-positive tumor-infiltrating lymphocytes and improved recurrence-free survival (P=0.041) was maintained only when the tumors had low-level PD-L1 expression (P=0.006); patients whose tumors had both high PD-1-positive tumor-infiltrating lymphocytes and high PD-L1 expression had a significantly worse recurrence-free survival (P<0.001). Thus, our results not only provide a foundation for further assessment of PD-L1 immunohistochemistry as a predictive marker for anti-PD-1 therapy in colorectal carcinoma, they also shed light on the prognostic impact of tumor-infiltrating lymphocytes in different subsets of mismatch-repair-deficient colorectal carcinomas.

Rab11 collaborates E-cadherin to promote collective cell migration and indicates a poor prognosis in colorectal carcinoma.

Collective cell migration, whereby the cell-cell contacts such as E-cadherin are maintained during migration, has only recently emerged, and its detailed mechanisms are still unclear. In this study, the role of Rab11, which functions in recycling endosomes, and its relationship to E-cadherin in colorectal carcinoma were identified, and the role of Rab11 in the collective cell migration of colon cancer cells was clarified. A total of 107 patients with surgically resected colorectal carcinoma were enrolled in this immunohistochemical study. Relationships between the overexpression of Rab11 and E-cadherin and survival were evaluated. The cell biology of Rab11 overexpression or knock-down in HT-29 colon cells was studied. The expression of Rab11 and E-cadherin was not correlated with the stage of cancer or lymph node metastasis. However, the overall survival was poor in the group of 67 patients with duo-positive Rab11 and E-cadherin expression compared to the group (40 patients) without dual-positive expression (P = 0·038). Rab11 was demonstrated to have a physical interaction with E-cadherin, and overexpression of Rab11 was found to promote collective cell migration through the increased distribution of E-cadherin, which enhanced cell-cell connections. In addition, Rac1 activation and matrix metalloproteinase-2 expressions were upregulated upon Rab11 expression. This study demonstrated that Rab11 and E-cadherin expressions are indicators of poor survival time in colorectal carcinoma, but that Rab11 overexpression may contribute to increased collective cell invasion in colorectal carcinoma.

Dysregulation of MALAT1 and miR-619-5p as a prognostic indicator in advanced colorectal carcinoma.

The present study aimed to detect the expression of metastasis associated lung adenocarcinoma transcript 1 (MALAT1) and microRNA (miR)-619-5p in colorectal carcinoma (CRC), and to evaluate the significance of MALAT1 and miR-619-5p expression in the clinical diagnosis and prognosis of CRC. Quantitative polymerase chain reaction was used to detect MALAT1 and miR-619-5p expression in 120 colorectal carcinoma and 120 adjacent normal tissue samples. The expression levels of MALAT1 and miR-619-5p were significantly different between colorectal carcinoma and adjacent normal tissues (P<0.05). MALAT1 exhibited an average 2.52-fold increase in colorectal adenoma when compared with adjacent normal tissues, while miR-619-5p exhibited an average 5.79-fold decrease in colorectal adenoma when compared with adjacent normal tissues. There was a significant difference between the MALAT1 expression in CRC tissues obtained from men and women (P=0.027), and in tumor-node-metastasis (TNM) stage II and stage III lesions (P=0.019). MALAT1 expression was associated with lymphovascular invasion (P=0.047) and perineural invasion (P=0.012). In addition, miR-619-5p expression was also significantly different between men and women (P=0.032), and between TNM stage II and stage III lesions (P=0.012). miR-619-5p expression was also associated with lymphovascular invasion (P=0.023) and perineural invasion (P=0.009). Patients with high expression of MALAT1 and low expression of miR-619-5p demonstrated significantly shorter disease-free survival (DFS) (P=0.002) and overall survival (OS) times (P=0.004) compared with patients with low MALAT1 expression and high miR-619-5p expression. Patients with perineural invasion demonstrated significantly shorter DFS (P=0.001) and OS times (P=0.003) compared with patients without perineural invasion. In addition, there was a negative correlation between MALAT1 expression and miR-619-5p expression (r=-0.415, P=0.004) in CRC tissues. In conclusion, MALAT1 and miR-619-5p have potential for the molecular diagnosis of CRC patients, and combined assaying of MALAT1 and miR-619-5p may improve the accuracy of the diagnosis of CRC and act as a good prognostic indicator in CRC patients.

Molecular profile of 5-fluorouracil pathway genes in colorectal carcinoma

BackgroundThis study addresses involvement of major 5-fluorouracil (5-FU) pathway genes in the prognosis of colorectal carcinoma patients.MethodsTesting set and two validation sets comprising paired tumor and adjacent mucosa tissue samples from 151 patients were used for transcript profiling of 15 5-FU pathway genes by quantitative real-time PCR and DNA methylation profiling by high resolution melting analysis. Intratumoral molecular profiles were correlated with clinical data of patients. Protein levels of two most relevant candidate markers were assessed by immunoblotting.ResultsDownregulation of DPYD and upregulation of PPAT, UMPS, RRM2, and SLC29A1 transcripts were found in tumors compared to adjacent mucosa in testing and validation sets of patients. Low RRM2 transcript level significantly associated with poor response to the first-line palliative 5-FU-based chemotherapy in the testing set and with poor disease-free interval of patients in the validation set irrespective of 5-FU treatment. UPP2 was strongly methylated while its transcript absent in both tumors and adjacent mucosa. DPYS methylation level was significantly higher in tumor tissues compared to adjacent mucosa samples. Low intratumoral level of UPB1 methylation was prognostic for poor disease-free interval of the patients (P = 0.0002). The rest of the studied 5-FU genes were not methylated in tumors or adjacent mucosa.ConclusionsThe observed overexpression of several 5-FU activating genes and DPYD downregulation deduce that chemotherapy naïve colorectal tumors share favorable gene expression profile for 5-FU therapy. Low RRM2 transcript and UPB1 methylation levels present separate poor prognosis factors for colorectal carcinoma patients and should be further investigated.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-016-2826-8) contains supplementary material, which is available to authorized users.