Effects of norcantharidin on the growth and apoptosis of breast cancer cell line MCF-7~(adr)

Objective:To explore the effect of norcantharidin (NCTD) on the growth and apoptosis of breast cancer cell line MCF-7adr for providing reliable evidence for clinical application.Methods:Suppressive effect of NCTD on cell line was detected by MTT assay. Morphological alterations were observed by light microscope and fluorescence microscope. Flow cytometry were used to observe the effect on cell cycle and apoptosis. The related factors of proliferation, apoptosis were determined by immunocytochemisty.Results:NCTD inhibited the growth and proliferation of MCF-7adr cells in a dose dependent manner and a time dependent manner. NCTD reduced the positive percentage of the ki-67 positive index from (96.8±1.3)% to(18.8±1.9)% at 48h after treatment; the percentage of PCNA was decreased from (95.4±1.1)% to(59.2±1.9)%.Morphological alterations of NCTD-treated cells under a microscope were membrane blebbing, condensation of nuclear chromatin, nuclear fragmentation, and apoptotic bodies. Hoechst staining assay confirmed this. Sub-G1 peaks were increasing in a dose dependent manner at 48h after NCTD treated cells. When dose of NCTD treated cells by 120、240、360 and 480μM, The increasing apoptosis ratio was 11.67%, 16.51%, 18.29% and 20.79%, respectively. The express ratio of p53 protein reduced from (97.8±1.64)% to (47.2±9.8)%. The express ration of bcl-2 protein decreased from (98.87±3.74)% to (17.5±1.64)%.Conclusion:This study shows the inhibition of NCTD on MCF-7adr cells. After NCTD treatment, cell apoptosis is promoted. The expression of apoptosis relative factors, mutant p53 is down-regulated.