title: Effect of HA14-1 on apoptosis-regulating proteins in HeLa cells.
abstract: Overexpression of Bcl-2 has been recognized in various malignancies. Recently, HA14-1, a Bcl-2 antagonist, has been identified for its anti-apoptotic effect. However, mode of action of HA14-1 still remains to be elucidated. In this study, we examined HA14-1 binding efficiency with receptor proteins through molecular docking. Cell viability using HeLa cells was evaluated through MTT assay after exposure to different concentration of HA14-1. Moreover, after HA14-1 exposure, expressions of tumor suppressor protein (p53), BH3-only protein (Puma) and apoptosis-associated proteins were analyzed by Western blotting. From the results, it was found that HA14-1 occupied all three domains; BH1, BH2, and BH3 within the hydrophobic pocket of Bcl-2. However, HA14-1 occupied only BH1 and BH3 of Bcl-xl, conversely, no such stable bond was observed for Bax and Bak. ARG107 and TYR101 were the amino acids involved in the binding of HA14-1 to Bcl-2 and Bcl-xl, respectively. Additionally, decrease in Bcl-2 and Bcl-xl expression along with increase in p53 and Puma expression after exposure to HA14-1 was observed. The results suggested p53 pathway to be the probable mechanism of action for the induction of apoptosis in HeLa cell by downregulating the effect of anti-apoptotic proteins suggesting that HA14-1 may provide therapeutic potential for the treatment of human cervical cancer.
authors.affiliation: Institute of Pharmacology, Toxicology and Biochemical Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Kanwal
authors.initials: K
authors.lastname: Rehman
authors.forename: Muhammad
authors.initials: M
authors.lastname: Tariq
authors.forename: Muhammad S H
authors.initials: MS
authors.lastname: Akash
authors.forename: Zeeshan
authors.initials: Z
authors.lastname: Gillani
authors.forename: Mehmood H
authors.initials: MH
authors.lastname: Qazi
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001578:Benzopyrans / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D001665:Binding Sites; D002470:Cell Survival / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D062105:Molecular Docking Simulation; D009570:Nitriles / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051020:bcl-X Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; C438378:BBC3 protein, human; D001578:Benzopyrans; D009570:Nitriles; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D016159:Tumor Suppressor Protein p53; D050998:bcl-2 Homologous Antagonist-Killer Protein; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; C456800:ethyl 2-amino-6-bromo-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate
keywords: anti-apoptotic proteins; apoptosis-regulating proteins; cervical cancer; pro-apoptotic proteins