Cytotoxicity of berberine on human cervical carcinoma HeLa cells through mitochondria, death receptor and MAPK pathways, and in-silico drug-target prediction.
Berberine, a natural product, has been widely used to treat hyperlipoidemia and intestinal diseases. In the present paper, berberine showed a significant anti-proliferative effect to human cervical carcinoma HeLa cells confirmed by 3-(4,5)-dimethyl-thiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT), flow cytometry analysis (FCM) and so on. The methods including western blotting, radioimmunity assay (RIA), reverse transcription-polymerase chain reaction (RT-PCR) were used to investigate protein and mRNA expressions. We found that Bcl-2/Bax ratio was significantly decreased and cytochrome c was released from mitochondrion to cytosol, which indicated that the mitochondrial pathway was activated by berberine. The up-regulation of Fas, FasL, TNF-alpha and TRAF-1 indicated the involvement of the death receptor pathway in the process of berberine-induced apoptosis. Furthermore caspase-3 and caspase-8 were activated as a central event of apoptosis, and the levels of phosphorylation of mitogen-activated protein kinases (MAPKs) were also investigated. In addition, the increased expression of p53 was also observed in berberine-treated HeLa cells, and as a node point of these different pathways in a protein-protein interaction network constructed by GeneGo software, p53 might be the possible drug-target of berberine's anti-cancer on HeLa cells, which was predicted by a flexible ligand-protein inverse docking program, INVDOCK. This study is benefit for clarifying the mechanism of berberine's anti-tumor effect and might be helpful to find therapy-target for treatment of human cervical carcinoma.
Lu
Binan
B
College of Pharmacy, Dalian Medical University, Dalian, China.
Hu
Mingming
M
Liu
Kexin
K
Peng
Jinyong
J
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001599:Berberine / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D019295:Computational Biology; D055808:Drug Discovery; D053222:Fas Ligand Protein / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D008928:Mitochondria / Q000187:drug effects*; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D053218:Receptors, Death Domain / Q000378:metabolism*; D047990:TNF Receptor-Associated Factor 1 / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism; D014410:Tumor Stem Cell Assay; D002583:Uterine Cervical Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D019014:fas Receptor / Q000235:genetics / Q000378:metabolism
D000970:Antineoplastic Agents; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D012333:RNA, Messenger; D053218:Receptors, Death Domain; D047990:TNF Receptor-Associated Factor 1; D014409:Tumor Necrosis Factor-alpha; D019014:fas Receptor; D001599:Berberine; D020929:Mitogen-Activated Protein Kinase Kinases