Norcantharidin induces apoptosis in human prostate cancer cells through both intrinsic and extrinsic pathways

Background norcantharidin,a modified pure compound from blister beetles,was previously demonstrated to induce apoptosis of cancer cells. this study investigated its anti-cancer activity in prostate cancer cells and the mechanisms involved. methods two human prostate cancer cell lines,22rv1 and du145,were treated with norcantharidin at concentrations ranging from 3 to 30 μg/ml. cytotoxic effect of norcantharidin was determined by use of the 3-(4,5-dimethylthiazol-yl)-diphenyl tetrazoliumbromide (mtt) assay. the effects of apoptosis were evaluated by cell death assay,caspase-3,-8,-9 activity and cytochrome c release. the apoptotic related protein expressions (bcl-2 family and inhibitor of apoptosis proteins) were determined using western blotting. results an mtt assay revealed that norcantharidin induced cytotoxicity against both prostate cancer cells in dose- and time-dependent manners. treatment with norcantharidin at 3 μg/ml or higher significantly increased oligonucleosomal formation with concomitant appearance of parp cleavage,implicating the induction of apoptosis. norcantharidin intrinsically elevated cytosolic cytochrome c levels and activated caspase-3,-8,and -9. extrinsically,it upregulated the expression of not only the death receptors fas and dr5 in 22rv1 cells,but also of rip and tradd adaptor proteins in du145 cells. mechanistically,norcantharidin increased ratios of pro-/anti-apoptotic proteins and decreased expression of iap family member proteins,including ciap1 and survivin,regardless of the distinct status of androgen receptor expression in both cells. conclusions norcantharidin exhibited cytotoxicity against 22rv1 and du145 prostate cancer cells by inducing both intrinsic and extrinsic apoptotic pathways and could thus potentially be a remedy for prostate cancer. © 2016 institute of pharmacology,polish academy of sciences. published by elsevier sp. z o.o. all rights reserved.