Minimally Invasive Urology Center, Shandong Provincial Hospital affiliated with Shandong University, Jinan, China.
Department of Urology, Shandong Provincial Hospital affiliated with Shandong University, Jinan, China.
Prostate Cancer Prostatic Dis. 2016 Dec;19(4):358-366. doi: 10.1038/pcan.2016.29. Epub 2016 Jul 19.
Berbamine (BBM) has been reported with antitumor activities. BBM inhibited the growth of prostate cancer (PCa) cells and caused vacuolization of mitochondria in preliminary study. We hypothesized BBM could enhance apoptosis of PCa cells through mitochondrial pathway.
Growth of PC-3 and LNCaP cells treated by BBM was determined by cell viability assay. The morphology of mitochondria was observed by electron microscopy. Apoptosis was quantified by flow cytometry. Expressions of bax/bcl-2, active caspase-9 and active caspase-3 were examined by western blot and real-time PCR. Methazolamide, an inhibitor of cytochrome c, was added to examine its blockade effect on BBM. PC-3 cells were injected subcutaneously into athymic mice, and BBM or saline was administrated intravenously. Diameters of induced tumors were compared, and ratios of apoptotic cells in tumor tissues were calculated.
BBM inhibited viability of cultured PC-3 and LNCaP cells in dose- and time-dependent manners. Flow cytometry showed BBM induced apoptosis in cultured cells. Mitochondria showed swelling, vacuolization and fused mitochondrial cristae. Western blot and real-time PCR analysis both showed increases of bax/bcl-2, active caspase-9 and active caspase-3. Methazolamide impeded effect of BBM on inducing apoptosis of cultured cells, and activations of caspase-9 and caspase-3 were also inhibited. Growth of tumors by grafted PC-3 cells was significantly slower in BBM group. Ratios of apoptotic cells in tumors were significantly higher in BBM group. Expressions of active caspase-9 and active caspase-3 in tumors were significantly upregulated by BBM.
BBM exhibited strong anti-PCa activities in vitro and in vivo relying on activating caspase-3 via intrinsic pathway of apoptosis, holding a great promise to develop new strategies for PCa.
小檗胺（BBM）具有抗肿瘤活性。在初步研究中，BBM 抑制前列腺癌（PCa）细胞的生长并导致线粒体空泡化。我们假设 BBM 可以通过线粒体途径增强 PCa 细胞的凋亡。
用细胞活力测定法测定 PC-3 和 LNCaP 细胞经 BBM 处理后的生长情况。用电子显微镜观察线粒体的形态。用流式细胞术定量检测细胞凋亡。用 Western blot 和实时 PCR 检测 bax/bcl-2、活性 caspase-9 和活性 caspase-3 的表达。加入甲噻唑胺（一种细胞色素 c 的抑制剂），以检查其对 BBM 的阻断作用。将 PC-3 细胞皮下注射到无胸腺小鼠中，静脉内给予 BBM 或生理盐水。比较诱导肿瘤的直径，并计算肿瘤组织中凋亡细胞的比例。
BBM 以剂量和时间依赖的方式抑制培养的 PC-3 和 LNCaP 细胞的活力。流式细胞术显示 BBM 诱导培养细胞凋亡。线粒体肿胀、空泡化和融合的线粒体嵴。Western blot 和实时 PCR 分析均显示 bax/bcl-2、活性 caspase-9 和活性 caspase-3 增加。甲噻唑胺阻碍了 BBM 对培养细胞凋亡的诱导作用，同时也抑制了 caspase-9 和 caspase-3 的激活。移植 PC-3 细胞的肿瘤生长在 BBM 组明显较慢。BBM 组肿瘤中凋亡细胞的比例明显较高。BBM 显著上调肿瘤中活性 caspase-9 和活性 caspase-3 的表达。
BBM 在体外和体内均表现出强大的抗 PCa 活性，依赖于通过细胞凋亡的内在途径激活 caspase-3，为开发治疗 PCa 的新策略提供了巨大的希望。
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