Integrated transcriptome and in vitro analysis revealed anti-proliferative effect of citral in human stomach cancer through apoptosis

Sri Renukadevi Balusamy,Sathishkumar Natarajan,Yeon Ju Kim,Sivasubramanian Ramani,Haribalan Perumalsamy

doi:10.1038/s41598-019-41406-8

Sri Renukadevi Balusamy, Sathishkumar Natarajan + Show 3 more

Open Access

https://doi.org/10.1038/s41598-019-41406-8

Copy DOI

Abstract

Cancer is the second leading cause of death globally, particularly stomach cancer is third most common causes of cancer death worldwide. Citral possesses anti-tumor activity in various cancer cell lines, However its effect toward stomach cancer and its mechanism of action is have yet to be elucidated. The goal of the present study is to elucidate the role of citral in stomach cancer using transcriptome and in vitro approaches. We performed transcriptome analysis using RNA-seq and explored its capability to persuade apoptosis in AGS human stomach cancer cell lines in vitro. Furthermore, the enrichment and KEGG pathway results suggested that there are several genes involved to induce apoptosis pathway. Furthermore, our study also demonstrated that citral arrested colony formation and migration of cancer cells significantly than that of untreated cells. RNA-seq revealed a total of 125 million trimmed reads obtained from both control and citral treated groups respectively. A total number of 612 differentially expressed genes (DEGs) were identified which includes 216 genes up-regulated and 396 genes down-regulated genes after treatment. The enrichment analysis identified DEGs genes from transcriptome libraries including cell death, cell cycle, apoptosis and cell growth. The present study showed the significant inhibition effect upon citral by regulating various genes involved in signaling pathways, inhibits metastasis, colony formation and induced apoptosis both in silico and in vitro.

Highlights

Stomach cancer is the third most among common cancer and second leading cause of death in the world[1,2]
This is the first report to combine both RNA seq and in vitro analysis to prove the effect of citral on suppressing stomach cancer growth by promoting apoptosis
The major active constituent citral was isolated identified through various spectroscopic analyses; including electron ionized mass spectrometry (EI-MS) and nuclear magnetic resonance (NMR) spectroscopy

Summary

Introduction

Stomach cancer is the third most among common cancer and second leading cause of death in the world[1,2]. Www.nature.com/scientificreports fruits rich in vitamin c prevent the development of gastric cancer by delaying tumor progression in experimental animals[10] These delaying actions may be due to any one of the combinational mechanisms: procarcinogen activation prevention, inactivating carcinogens, stimulating DNA repair mechanisms, down-regulating protooncogene expression, up-regulation of tumor suppressor genes, inhibition of cell proliferation, differentiation induction and promoting apoptosis, stimulation of immune system, and regulating transcription factors and abnormal signaling pathways[11,12]. There are several previous studies demonstrated the role of citral on inhibiting the growth of cancer cells by targeting molecular signaling pathways including small-cell lung cancer; breast cancer-MDA-MB-231; Colorectal cancer[15,16] This is the first report to combine both RNA seq and in vitro analysis to prove the effect of citral on suppressing stomach cancer growth by promoting apoptosis

Objectives

Methods

Results