Bufalin alters gene expressions associated DNA damage, cell cycle, and apoptosis in human lung cancer NCI-H460 cells in vitro.
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.
Institute of Clinical Medical Science, China Medical University, Taichung 40402, Taiwan.
Shin-Hwar
SH
Wu
Department of Biological Science and Technology, China Medical University, Taichung 40402, Taiwan.
Yung-Ting
YT
Hsiao
Department of Medicinal Botany and Health Applications, Da-Yeh University, Changhua 51591, Taiwan.
Jaw-Chyum
JC
Chen
Department of Biological Science and Technology, China Medical University, Taichung 40402, Taiwan.
Ju-Hwa
JH
Lin
Department of Biological Science and Technology, China Medical University, Taichung 40402, Taiwan.
Shu-Chun
SC
Hsu
Gradualted Institute of Chinese Medical Science, China Medical University, Taichung 40402, Taiwan.
Te-Chun
TC
Hsia
Department of Radiology, China Medical University Hospital, Taichung 40447, Taiwan.
Su-Tso
ST
Yang
Department of Internal Medicine, China Medical University Hospital, Taichung 40447, Taiwan. hsuwh@mail.cmuh.org.tw.
Wu-Huei
WH
Hsu
Department of Biological Science and Technology, China Medical University, Taichung 40402, Taiwan. jgchung@mail.cmu.edu.tw.
Jing-Gung
JG
Chung
D017209:Apoptosis / Q000187:drug effects*; D002018:Bufanolides / Q000008:administration & dosage*; D002453:Cell Cycle / Q000187:drug effects*; D004249:DNA Damage / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000473:pathology
D002018:Bufanolides; C022777:bufalin