Genistein Modulates Signaling Pathways and Targets Several Epigenetic Markers in HeLa Cells.

Madhumitha Kedhari Sundaram,Arif Hussain,Raju K Mandal,Shafiul Haque,Sreepoorna Unni,Tulika Bhardwaj,Pallavi Somvanshi

doi:10.3390/genes10120955

Abstract

Background: Several epigenetic changes are responsible for transcriptional alterations of signaling pathways and tumour suppressor genes (TSGs) contributing to carcinogenesis. This study was aimed to examine the effect of the phytochemical, genistein on various molecular targets in HeLa cells. Methods: Quantitative PCR was used to analyze the expression of various molecular targets. Biochemical assays were employed to study the epigenetic enzymes. To correlate the transcriptional status of the selected TSGs and epigenetic modulation, their promoter 5’CpG methylation levels were evaluated by quantitative methylation array followed by methylation specific restriction digestion. Results: The expression of several genes involved in the cell cycle regulation, migration, inflammation, phosphatidylinositol 3-kinase (PI3K) and mitogen activated kinase-like protein (MAPK) pathway were found to be modulated including CCNB1, TWIST1, MMP14, TERT, AKT1, PTPRR, FOS and IL1A. Genistein modulated the expression of DNA methyltransferases (DNMTs), histone deacetylases (HDACs), histone methyltransferases (HMTs), demethylases, and histone phosphorylases. Furthermore, genistein decreased the activity of DNMTs, HDACs, and HMTs and reduced global DNA methylation levels. Promoter methylation of several TSGs, including FHIT, RUNX3, CDH1, PTEN, and SOC51, was lowered with corresponding transcriptional increase. Network analysis indicated similar effect of genistein. Conclusion: This study presents a comprehensive mechanism of action of genistein showcasing effective epigenetic modulation and widespread transcriptional changes resulting in restoration of tumour suppressor gene expression. This study corroborates the development of genistein as a candidate for anti-cancer therapy.

Highlights

Carcinogenesis involves both genetic and epigenetic changes, which can alter the expression of genes that are central to the development of cancer
Carcinogenesis is the fallout of aberrant changes in signal transduction pathways fueled by both genetic and epigenetic alterations
Epigenetic alterations are increasingly being acknowledged as an early and ongoing alteration that aids in cancer initiation and progression

Summary

Introduction

Carcinogenesis involves both genetic and epigenetic changes, which can alter the expression of genes that are central to the development of cancer. DNA methyltransferases (DNMTs), HDACs and other histone modifiers are found to be overexpressed in various cancers, changing the subtle equilibrium and allowing for tumor development and progression. The changes in the levels of promoter methylation and histone acetylation of tumor suppressor genes (TSGs) are an outcome of the disturbance in enzyme equilibrium and results in cancer development [8]. Several epigenetic changes are responsible for transcriptional alterations of signaling pathways and tumour suppressor genes (TSGs) contributing to carcinogenesis. Conclusion: This study presents a comprehensive mechanism of action of genistein showcasing effective epigenetic modulation and widespread transcriptional changes resulting in restoration of tumour suppressor gene expression. This study corroborates the development of genistein as a candidate for anti-cancer therapy

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Conclusion