Bufalin alters gene expressions associated DNA damage, cell cycle, and apoptosis in human lung cancer NCI-H460 cells in vitro.

Shin-Hwar Wu,Yung-Ting Hsiao,Jaw-Chyum Chen,Te-Chun Hsia,Shu-Chun Hsu,Wu-Huei Hsu,Jing-Gung Chung,Ju-Hwa Lin,Su-Tso Yang

doi:10.3390/molecules19056047

Abstract

Lung cancer is the leading cause of cancer related death and there is no effective treatment to date. Bufalin has been shown effective in inducing apoptosis and DNA damage in lung cancer cells. However, the genetic mechanisms underlying these actions have not been elucidated yet. Cultured NCI-H460 cells were treated with or without 2 μM of bufalin for 24 h. The total RNA was extracted from each treatment for cDNA synthesis and labeling, microarray hybridization, and then followed by flour-labeled cDNA hybridized on chip. The localized concentrations of fluorescent molecules were detected and quantitated and analyzed by Expression Console software (Affymetrix) with default RMA parameters. The key genes involved and their possible interaction pathways were mapped by GeneGo software. About 165 apoptosis-related genes were affected. CASP9 was up-regulated by 5.51 fold and THAP1 by 2.75-fold while CCAR1 was down-regulated by 2.24 fold. 107 genes related to DNA damage/repair were affected. MDC1 was down-regulated by 2.22-fold, DDIT4 by 2.52 fold while GADD45B up-regulated by 3.72 fold. 201 genes related to cell cycles were affected. CCPG1 was down-regulated by 2.11 fold and CDCA7L by 2.71 fold. Many genes about apoptosis, cell cycle regulation and DNA repair are changed significantly following bufalin treatment in NCI-H460 cells. These changes provide an in depth understanding of cytotoxic mechanism of bufalin in genetic level and also offer many potentially useful biomarkers for diagnosis and treatment of lung cancer in future.

Highlights

Non-small cell lung carcinoma is one of leading causes of cancer-related death in the United States and throughout the world [1]
The Up-Regulated and Down-Regulated Gene Expression in H460 Exposed to Bufalin
One hundred and seven affected genes are associated with DNA damage and repair, such as MDC1, that was down-regulated by 2.22-fold or DDIT4 (DNA-damage-inducible transcript 4) which was suppressed by 2.52-fold, whereas, GADD45B

Summary

Introduction

Non-small cell lung carcinoma is one of leading causes of cancer-related death in the United States and throughout the world [1] It causes more than one million deaths every year [2]. Natural products were the main source of health care in ancient times In modern medicine, they are still major sources of new drug development. Some oncogenic mutations involving inactivating apoptosis or disrupting DNA repair mechanism These vulnerable genes are targets of detection or treatment for various cancers. We are closer to understand the mechanisms underlying bufalin’s anti-cancer effects at the genetic level. We hope this knowledge will provide a basis for future studies on anti-cancer drug development or modification

Results and Discussion

Discussion

Chemicals and Reagents

Lung Cancer Cells

Conclusions