Prognostic Significance of Cytokeratin-20 Reverse Transcriptase Polymerase Chain Reaction in Lymph Nodes of Node-Negative Colorectal Cancer Patients

The aim of this study was to evaluate the association of single-nucleotide variant n.60G>C (rs2910164) of microRNA (miR)-146a, related to suppressing of BRCA1/2 DNA repair protein, with the risk and survival of colorectal cancer patients, as well as miR-146a and BRCA1/2 levels and miR binding efficiency. The genotypes were identified in 125 colorectal cancer patients and 276 controls using TaqMan polymerase chain reaction assay. The miR-146a and BRCA1/2 levels were assessed by quantitative–polymerase chain reaction protocols. Primary precursor of miR-146a containing G (wild-type) and C (variant) allele were cloned into pcDNA.3.3 vector and co-transfected in HT-29 colorectal cancer cell line. Luciferase reporter assay was performed to assess miR-146a binding to BRCA2 3′-untranslated region in HT-29. The differences between groups were calculated using chi-square or Fisher’s exact test, logistic regression, and Mann–Whitney test. The prognostic impact of single-nucleotide variant genotypes on overall survival was evaluated by Kaplan–Meier estimate and Cox regression. The GC or CC genotypes prevalence was similar in patients and controls (50.4% vs 50.7%, p = 0.74). However, patients with tumors in advanced stage with miR-146a GG genotype had 2.41 more chance of dying than GC or CC genotypes. In addition, tumor tissues of patients with GG genotype presented higher miR-146a ( p = 0.02) and lower BRCA1 ( p = 0.01) and BRCA2 ( p < 0.0001) levels when compared to those with GC or CC genotypes. In fact, pcDNA.3.3-miR-146a-G presented increased binding capacity to the 3′-untranslated region of BRCA2 ( p = 0.001) compared to pcDNA.3.3-miR-146a-C. In addition, the G allele altered the binding affinity between miR-146a and its BRCA2 3′-untranslated region target ( p < 0.001), thus enhancing suppression of BRCA2 expression. Our results suggest that single-nucleotide variant rs2910164 does not influence the colorectal cancer risk in Brazilian patients; however, the GG genotype could act as a factor of worse prognosis in patients with advanced disease due to suppression of BRCA1/2 modulated by miR-146a.

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Prognostic Significance of Cytokeratin-20 Reverse Transcriptase Polymerase Chain Reaction in Lymph Nodes of Node-Negative Colorectal Cancer Patients

Journal of Clinical Oncology ◽

10.1200/jco.2002.20.4.1049 ◽

2002 ◽

Vol 20 (4) ◽

pp. 1049-1055 ◽

Cited By ~ 54

Author(s):

Robert Rosenberg ◽

Axel Hoos ◽

James Mueller ◽

Patricia Baier ◽

Dominik Stricker ◽

...

Keyword(s):

Colorectal Cancer ◽

Overall Survival ◽

Lymph Nodes ◽

Prognostic Value ◽

Tumor Recurrence ◽

Cytokeratin 20 ◽

Rt Pcr ◽

Chain Reaction ◽

Node Negative ◽

Polymerase Chain

PURPOSE: Approximately 20% to 30% of patients with curatively resected, node-negative (pN0) colorectal cancer die of tumor recurrence, which can be caused by minimal residual disease. To identify patients with an increased risk of tumor recurrence and evaluate the prognostic value of cytokeratin-20 (CK-20), we detected CK-20–positive cells in histopathologically tumor-free lymph nodes (pN0) of patients with colorectal cancer. PATIENTS AND METHODS: Two peritumoral lymph nodes each from 85 patients with completely resected (R0) colorectal cancer without lymph node metastases (pN0) by routine examination were analyzed using a CK-20–specific reverse transcriptase polymerase chain reaction (RT-PCR) and compared with CK-20–specific immunohistochemistry (IHC). The results were correlated with histopathologic findings and with survival. RESULTS: CK-20 RT-PCR was positive in 44 patients (52%) and detected 83% of cancer-related death. Positive RT-PCR was significantly correlated with poorer overall survival (P < .009). Comparing RT-PCR with IHC, 13 patients with positive RT-PCR were identified, where the CK-20 expression was caused by tumor cell contamination located exclusively outside the lymph node capsule and had no prognostic impact. Defining these 13 patients as RT-PCR negative improved specificity of the RT-PCR assay from 57% to 75%. The 5-year overall survival of the 31 RT-PCR–positive patients was 71%, compared with 96% in the 54 negative patients (P < .001). Multivariate analysis showed expression of CK-20 mRNA to be an independent prognostic factor with a relative risk of cancer-related death of 6.1. CONCLUSION: CK-20 RT-PCR in peritumoral histopathologic tumor-free (pN0) lymph nodes of colorectal cancer is an independent prognostic factor for overall survival. Additional CK-20 IHC improves the specificity and prognostic value of RT-PCR for cancer-related death.

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Detection of colonic cells in peripheral blood of colorectal cancer patients by means of reverse transcriptase and polymerase chain reaction

British Journal of Cancer ◽

10.1038/bjc.1998.686 ◽

1998 ◽

Vol 78 (10) ◽

pp. 1368-1372 ◽

Cited By ~ 57

Author(s):

A Castells ◽

L Boix ◽

X Bessa ◽

L Gargallo ◽

JM Piqué

Keyword(s):

Colorectal Cancer ◽

Polymerase Chain Reaction ◽

Reverse Transcriptase ◽

Cancer Patients ◽

Peripheral Blood ◽

Chain Reaction ◽

Polymerase Chain ◽

Colorectal Cancer Patients ◽

Colonic Cells

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Genetic Polymorphisms of FOXO3a Gene in Colorectal Cancer among North Indian Population

10.21203/rs.3.rs-222413/v1 ◽

2021 ◽

Author(s):

Laraib Uroog ◽

Zafar Iqbal Bhat ◽

Bushra Zeya ◽

Khalid Imtiyaz ◽

Rauf AHMAD Wani ◽

...

Keyword(s):

Colorectal Cancer ◽

Polymerase Chain Reaction ◽

Cancer Patients ◽

Statistical Significance ◽

Incidence Rates ◽

Risk Haplotype ◽

Chain Reaction ◽

Increased Risk ◽

Polymerase Chain ◽

Colorectal Cancer Patients

Abstract Background Colorectal cancer (CRC) heritability is determined by the composite relations between inherited variants and environmental factors. In developing countries like India CRC incidence rates have been increasing specially. In the present study, we focused on the distribution of FOXO3a gene polymorphisms in North Indian colorectal cancer patients. Methods A case–control study was conducted on 900 samples including 450 colorectal cancer patients and 450 age matched controls. We genotyped the SNPs rs2253310 and rs4946936 via Polymerase Chain Reaction-Restriction fragment length polymorphism (RFLP) analysis and Polymerase Chain Reaction- single stranded conformation polymorphism (SSCP) procedure followed by sequence detection. Results A significantly increased risk of CRC was observed with rs4946936 genotype (P= 0.0393; OR= 1.405 CI=1.051-1.879). GT haplotype although not reaching statistical significance appeared to be at higher “risk” haplotype (OR- 1.164, 95%CI= 0.967~1.401), while as other haplotypes CC (OR- 0.893, 95% CI=0.665~1.200]), CT (OR- 0.806, 95%CI= 0.616~1.055) and GC (OR- 0.994, 95%CI= 0.809~1.221) were found to be “protective” for developing colorectal cancer. Conclusion This study lends support for an increased risk of CRC associated with the rs4946936 polymorphism. Nevertheless, statistically significant association between rs2253310 genotypes and CRC risk was not observed.

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