Human colon cancer cells lacking Bax resist curcumin-induced apoptosis and Bax requirement is dispen.pdfVIP

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Human colon cancer cells lacking Bax resist curcumin-induced apoptosis and Bax requirement is dispen.pdf

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Human colon cancer cells lacking Bax resist curcumin-induced apoptosis and Bax requirement is dispen

Human colon cancer cells lacking Bax resist curcumin-induced apoptosis and Bax requirement is dispensable with ectopic expression of Smac or downregulation of Bcl-XL Ramachandran Rashmi, Santhosh Kumar and Devarajan Karunagaran Cancer Biology Laboratory, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala 695 014, India To whom correspondence should be addressed. Tel: t91 471 2347975; Fax: t91 471 2348096; E-mail: dkarunagaran@ Multiple apoptotic stimuli induce conformational changes in Bax, a proapoptotic protein from the Bcl-2 family and its deficiency is a frequent cause of chemoresistance in colon adenocarcinomas. Curcumin, a dietary compound from turmeric, is known to induce apoptosis in a variety of cancer cells. To understand the role of Bax in curcumin- induced apoptosis we used HCT116 human colon cancer cells with one allele of Bax gene (Baxt/) and Bax knockout HCT116 (Bax/) cells in which Bax gene is inactivated by homologous recombination. Cell viability decreased in a concentration-dependent manner in Baxt/ cells treated with curcumin (0--50 mM) whereas only minimal changes in viability were observed in Bax/ cells upon curcumin treatment. In Bax/ cells curcumin-induced activation of caspases 9 and 3 was blocked and that of caspase 8 remained unaltered. Curcumin-induced release of cyto- chrome c, Second mitochondria derived activator of cas- pase (Smac) and apoptosis inducing factor (AIF) was also blocked in Bax/ cells and reintroduction of Bax, down- regulation of the antiapoptotic protein Bcl-XL by antisense DNA as well as the overexpression of Smac, highly sensit- ized the Bax/ cells toward curcumin-induced apoptosis. There was no considerable difference in the percentage of apoptotic cells in Bak RNAi transfected Baxt/ or Bax/ cells treated with curcumin when compared with their corresponding vector transfected cells treated with curcu- min. The present study demonstrates the role of Bax but not Bak as a critical regulat