Case Control Study
Copyright ©The Author(s) 2018. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Oct 21, 2018; 24(39): 4462-4471
Published online Oct 21, 2018. doi: 10.3748/wjg.v24.i39.4462
Molecular evaluation of glutathione S transferase family genes in patients with sporadic colorectal cancer
Gabriela Helena Rodrigues-Fleming, Glaucia Maria de Mendonça Fernandes, Anelise Russo, Patrícia Matos Biselli-Chicote, João Gomes Netinho, Érika Cristina Pavarino, Eny Maria Goloni-Bertollo
Gabriela Helena Rodrigues-Fleming, Glaucia Maria de Mendonça Fernandes, Anelise Russo, Patrícia Matos Biselli-Chicote, Érika Cristina Pavarino, Eny Maria Goloni-Bertollo, Genetics and Molecular Biology Research Unit - UPGEM, São José do Rio Preto Medical School, FAMERP, São José do Rio Preto, SP 15090-000, Brazil
João Gomes Netinho, Department of Surgery and Coloproctology, São José do Rio Preto Medical School, FAMERP, São José do Rio Preto, SP 15090-000, Brazil
Author contributions: Rodrigues-Fleming GH planned and conducted the study, collected and interpreted data, and drafted and wrote the manuscript; Fernandes GMM participated in the collection of the genetic material, performed the analytical assessments, and revised the manuscript; Russo A participated in the collection of the genetic material; Biselli-Chicote PM critically revised the analytical tools and the manuscript; Netinho JG collected data on sporadic colorectal cancer patients; Pavarino ÉC served as scientific advisor; Goloni-Bertollo EM was the guarantor, planned the study, and critically revised the manuscript.
Supported by the São Paulo Research Foundation (FAPESP), No. 2011/23969-1 and No. 2012/02473-0; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001 (Master Grant); and National Council of Technological and Scientific Development (CNPq) No. 310582/2014-8.
Institutional review board statement: The study was reviewed and approved by the FAMERP Institutional Review Board, and it was approved by the Ethics in Research Committee CEP/FAMERP, protocol no. 012/2012.
Informed consent statement: All patients gave informed consent.
Conflict-of-interest statement: The authors declare no conflicts of interest.
Data sharing statement: Participants gave written informed consent for data sharing.
STROBE statement: The authors have read the STROBE Statement-checklist of items, and the manuscript was prepared and revised according to the STROBE Statement.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Eny Maria Goloni-Bertollo, PhD, Adjunct Professor, Postdoc, Genetics and Molecular Biology Research Unit - UPGEM, Department of Molecular Biology, São José do Rio Preto Medical School (FAMERP), Av. Brigadeiro Faria Lima, - 5416 - Vila São Pedro, São José do Rio Preto, SP 15090-000, Brazil. eny.goloni@famerp.br
Telephone: +55-17-32015720 Fax: +55-17-32015708
Received: June 29, 2018
Peer-review started: July 2, 2018
First decision: July 17, 2018
Revised: July 27, 2018
Accepted: August 24, 2018
Article in press: August 24, 2018
Published online: October 21, 2018
Processing time: 111 Days and 20.7 Hours
ARTICLE HIGHLIGHTS
Research background

Colorectal cancer is the third most common cancer worldwide and develops on the inner walls of the colon and rectum. Genetic and environmental factors may increase the risk of colorectal cancer via the metabolism of carcinogens. Therefore, the evaluation of polymorphisms in genes involved in this process may help to modulate the development of colorectal cancer. Polymorphisms in genes encoding GSTP1, GSTT1, and GSTM1 may alter enzymatic activity. This change can lead to DNA damage and deregulation of the mechanisms involved in colorectal cancer.

Research motivation

Polymorphisms in the coding genes GSTP1, GSTT1, and GSTM1 have been studied in terms of susceptibility to diseases such as cancer. However, the literature presents conflicting results. Therefore, several studies are needed to assess and confirm the role of factors that influence changes in metabolic processes related to colorectal cancer.

Research objectives

The main objective of this study was to evaluate the influence of polymorphisms in the GSTP1, GSTT1 and GSTM1 genes on the risk of colorectal cancer. The data showed that carriers of polymorphisms in the GSTM1 genes and the combination of GSTT1 non-null/GSTM1 null genotypes and GSTT1 non-null/GSTM1 null/GSTP1 Val* (*with the presence of at least one polymorphic allele) constitute a risk group for sporadic colorectal cancer (SCRC), and polymorphisms in the GSTM1 gene and the GSTT1 non-null/GSTM1 null combinations, GSTT1 non-null/GSTM1 null/GSTP1 Val* increase the aggressiveness of the tumor. Thus, considering the high incidence of this cancer, it is important to understand the factors that lead to carcinogenesis in order to develop preventive and therapeutic strategies for the management of cancer.

Research methods

This case-control study evaluated 970 individuals, 232 cases and 738 controls, using multiplex polymerase chain reaction (PCR) and polymerase chain reaction-restriction fragment chain reaction (PCR-RFLP) polymorphism, Demographics are tabulated by percentage. The binary logistic regression model was used to evaluate the association of age, gender, smoking and eating habits with SCRC, and to evaluate the association of the Hardy-Weinberg equilibrium (HWE) with the Chi-square test. Multiple binary logistic regression, adjusted for age, gender and smoking and alcohol habits, was also used to evaluate the association between the genetic models of the polymorphisms and the development of SCRC. The dominant genotypic model was used to assess the interaction between polymorphisms and smoking habits, adjusted for age, gender, and ethnicity, and to evaluate the interaction of polymorphisms and alcohol consumption, adjusted for age, gender and smoking, on the risk of SCRC. In addition, the Kapla-Meier curve was used to assess the overall survival of patients with SCRC.

Research results

The data showed that carriers of polymorphisms in the GSTM1 genes and the combination of GSTT1 non-null/GSTM1 null genotypes and GSTT1 non-null/GSTM1 null/GSTP1 Val* (*with the presence of at least one polymorphic allele) constitute a risk group for SCRC, and polymorphisms in the GSTM1 gene and the GSTT1 non-null/GSTM1 null combinations, GSTT1 non-null/GSTM1 null/GSTP1 Val* increase the aggressiveness of the tumor. Thus, considering the high incidence of this cancer, it is important to understand the factors that lead to carcinogenesis in order to develop preventive and therapeutic strategies for the management of cancer.

Research conclusions

Similar studies have not previously been performed in the Brazilian population. Therefore, this work is unprecedented in this population. In addition, we emphasize the importance of the association between female gender and susceptibility to SCRC as well as the survival analysis associated with the polymorphisms studied, which have not been extensively studied in the literature. Polymorphisms in the GSTP1, GSTT1 and GSTM1 genes were involved in carcinogenesis and the poor prognosis of SCRC. In the Brazilian population it was observed that females with advanced age were more susceptible to SCRC. The presence of the GSTM1 null genotype, and the combination of GSTT1/GSTM1 and GSTT1/GSTM1/GSTP1 genotypes are associated with an increased risk of SCRC and tumor progression.

This study provides a perspective on biomarkers of GSTs related to the prognosis of SCRC that has not been extensively studied in other populations, especially the Brazilian population. These data may contribute to clinical practice. Another interesting fact was the association between females, age over 60 years and the risk of SCRC. Menopausal women (estrogen reduction) were also shown to be more susceptible to SCRC. Polymorphisms in the genes involved in the xenobiotic metabolism pathway are associated with the development and poor prognosis of SCRC.

In this study, statistical analyses were widely used, and unlike other studies, multiple logistic regression was performed to evaluate the interactions between the polymorphisms studied and variables. In addition, survival was assessed by Kaplan Meier analysis. These analyses are extremely relevant in studies involving population genetic polymorphisms.

An association between some polymorphisms of xenobiotic metabolism and the development and progression of SCRC was observed. Advanced age and female gender were associated with the development of SCRC and polymorphisms in the genes involved in the xenobiotic metabolism pathway were associated with the development and poor prognosis of SCRC. This study may contribute to GSTs being used as diagnostic and prognostic biomarkers for SCRC. These data together with the findings of other studies may contribute to the development of treatment strategies for SCRC.

Research perspectives

This study demonstrated the importance of population studies with a large sample size in research on polymorphisms. Thus, we intend to expand the sample size to validate the results and include more polymorphisms related to the xenobiotic pathways in order to better understand the roles of these pathways in SCRC carcinogenesis. Research methods such as real-time PCR, are important in order to accurately quantify the presence of polymorphisms.