Alternative Carcinogenicity Screening Assay Using Colon Cancer Stem Cells: A Quantitative PCR (qPCR)-Based Prediction System for Colon Carcinogenesis

J Microbiol Biotechnol. 2018 Apr 28;28(4):645-651. doi: 10.4014/jmb.1712.12043.

Abstract

The carcinogenicity of chemicals in the environment is a major concern. Recently, numerous studies have attempted to develop methods for predicting carcinogenicity, including rodent and cell-based approaches. However, rodent carcinogenicity tests for evaluating the carcinogenic potential of a chemical to humans are time-consuming and costly. This study focused on the development of an alternative method for predicting carcinogenicity using quantitative PCR (qPCR) and colon cancer stem cells. A toxicogenomic method, mRNA profiling, is useful for predicting carcinogenicity. Using microarray analysis, we optimized 16 predictive gene sets from five carcinogens (azoxymethane, 3,2'-dimethyl-4-aminobiphenyl, N-ethyl-n-nitrosourea, metronidazole, 4-(n-methyl-n-nitrosamino)-1-(3-pyridyl)-1-butanone) used to treat colon cancer stem cell samples. The 16 genes were evaluated by qPCR using 23 positive and negative carcinogens in colon cancer stem cells. Among them, six genes could differentiate between positive and negative carcinogens with a p-value of < or =0.05. Our qPCR-based prediction system for colon carcinogenesis using colon cancer stem cells is cost- and time-efficient. Thus, this qPCR-based prediction system is an alternative to in vivo carcinogenicity screening assays.

Keywords: Microarray; cancer stem cell; carcinogenicity; colon cancer; quantitative PCR.

MeSH terms

  • Biomarkers, Tumor / analysis
  • Biomarkers, Tumor / genetics
  • Carcinogenesis / genetics*
  • Carcinogenicity Tests / economics
  • Carcinogenicity Tests / methods*
  • Carcinogens / toxicity*
  • Cell Survival / drug effects
  • Colon
  • Colonic Neoplasms / chemically induced*
  • Colonic Neoplasms / genetics*
  • DNA Primers
  • Gene Expression Profiling / methods
  • Gene Expression Regulation, Neoplastic / genetics
  • Gene Regulatory Networks
  • HCT116 Cells
  • Humans
  • In Vitro Techniques
  • Neoplastic Stem Cells*
  • RNA, Messenger / analysis
  • Real-Time Polymerase Chain Reaction / economics
  • Real-Time Polymerase Chain Reaction / methods*
  • Time Factors

Substances

  • Biomarkers, Tumor
  • Carcinogens
  • DNA Primers
  • RNA, Messenger