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Effect of HA 14‐1 on Apoptosis‐Regulating Proteins in HeLa Cells
Author(s) -
Rehman Kanwal,
Tariq Muhammad,
Akash Muhammad S. H.,
Gillani Zeeshan,
Qazi Mehmood H.
Publication year - 2014
Publication title -
chemical biology and drug design
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.59
H-Index - 77
eISSN - 1747-0285
pISSN - 1747-0277
DOI - 10.1111/cbdd.12245
Subject(s) - hela , puma , apoptosis , blot , mtt assay , viability assay , chemistry , docking (animal) , microbiology and biotechnology , cell , biology , biochemistry , gene , medicine , nursing
Overexpression of Bcl‐2 has been recognized in various malignancies. Recently, HA 14‐1, a Bcl‐2 antagonist, has been identified for its anti‐apoptotic effect. However, mode of action of HA 14‐1 still remains to be elucidated. In this study, we examined HA 14‐1 binding efficiency with receptor proteins through molecular docking. Cell viability using HeLa cells was evaluated through MTT assay after exposure to different concentration of HA 14‐1. Moreover, after HA 14‐1 exposure, expressions of tumor suppressor protein (p53), BH 3‐only protein (Puma) and apoptosis‐associated proteins were analyzed by Western blotting. From the results, it was found that HA 14‐1 occupied all three domains; BH 1, BH 2, and BH 3 within the hydrophobic pocket of Bcl‐2. However, HA 14‐1 occupied only BH 1 and BH 3 of Bcl‐xl, conversely, no such stable bond was observed for Bax and Bak. ARG 107 and TYR 101 were the amino acids involved in the binding of HA 14‐1 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 HA 14‐1 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 HA 14‐1 may provide therapeutic potential for the treatment of human cervical cancer.