Upregulation of hyaluronan binding protein 1 (HABP1/p32/gC1qR) is associated with Cisplatin induced apoptosis.
We have earlier reported that overexpression of HABP1 in fibroblast cells causes perturbed cell growth, extensive vacuolation and restricted entry to the S-phase, finally leading to apoptosis (Biochem Biophys Res Commun 2003; 300: 686-693). In the present study, we investigate the regulation of HABP1 expression in cisplatin induced apoptosis in HeLa cells. Apoptosis induced in HeLa cells at 24 h of cisplatin treatment was confirmed by nuclear fragmentation, increase in subdiploid population and the enhanced activation of ERK and upregulation of p53. In association with apoptosis induction, an upregulation of HABP1 expression was observed in HeLa cells at 18 and 24 h of cisplatin treatment. Quantification of HABP1 expression by flow cytometry confirmed a two-fold increase in total intracellular HABP1 expression at 24 h of cisplatin treatment. Under the same condition the HABP1 transcript level measured by semi quantitative RT PCR showed 2.5-fold increase ascertaining transcriptional regulation of HABP1 during cisplatin induced apoptosis. Further, in normal HeLa cells though a small amount of HABP1 can be detected in nucleus, but with apoptosis induction the protein is mainly concentrating around the nuclear periphery at 18 h of cisplatin treatment and is present both in the nucleus as well as in the cytosol at 24 h of treatment, suggesting its nuclear translocation during apoptosis. To substantiate our findings prior to the cisplatin treatment, the expression of HABP1 was reduced by small interfering RNA mediated knockdown. We observed a reduction in apoptotic cell population in cisplatin treated HeLa cells with disrupted HABP1 conferring resistance to cisplatin induced apoptosis. We report here that HABP1 upregulation in the cell is important for cisplatin induced apoptosis.
Biochemistry Laboratory, School of Environmental Sciences, Jawaharlal Nehru University, New Delhi-110067, India.
Anupama
A
Kamal
K
K
Datta
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002352:Carrier Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002945:Cisplatin / Q000494:pharmacology*; D004789:Enzyme Activation; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D024101:Mitochondrial Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000378:metabolism; D015854:Up-Regulation
D000970:Antineoplastic Agents; C087299:C1QBP protein, human; D002352:Carrier Proteins; D024101:Mitochondrial Proteins; D016159:Tumor Suppressor Protein p53; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D002945:Cisplatin