基于线粒体凋亡通路探讨防己诺林碱对前列腺癌PC3细胞的生物学行为的影响

Effects of fangchinoline on biological behavior of prostate cancer PC3 cells based on mitochondrial apoptosis pathway

目的 基于线粒体凋亡通路探究防己诺林碱(fangchinoline, FAN)对前列腺癌PC3细胞生物学行为的影响。方法 将前列腺癌PC3细胞按照FAN处理浓度分为4组,分别为FAN 0μmol/L组、FAN 5μmol/L组、FAN 10μmol/L组和FAN 20μmol/L组,各组依次采用FAN 0μmol/L (DMSO替代)、5μmol/L、10μmol/L和20μmol/L进行处理,孵育48 h后进行实验。CCK-8法检测PC3细胞增殖,平板克隆法检测PC3细胞克隆形成能力,流式细胞仪检测PC3细胞凋亡,Transwell实验检测PC3细胞侵袭,划痕实验检测PC3细胞迁移,Western blot检测PC3细胞自噬相关蛋白和线粒体凋亡通路蛋白。结果 与FAN 0μmol/L组相比,FAN 5μmol/L组、FAN 10μmol/L组和FAN 20μmol/L组的PC3细胞增殖能力、克隆形成能力、侵袭和迁移能力明显降低(P<0.001),凋亡率明显升高(P<0.001);与FAN 5μmol/L组和FAN 10μmol/L组相比,FAN 20μmol/L组的PC3细胞增殖能力、克隆形成能力、侵袭和迁移能力明显降低(P<0.001);与FAN 0μmol/L组相比,FAN 5μmol/L组、FAN 10μmol/L组和FAN 20μmol/L组的PC3细胞中P62和Bcl-2蛋白表达明显降低(P<0.001),LC3B、Bax和Caspase-3蛋白表达明显升高(P<0.001)。结论 FAN可能通过线粒体凋亡通路抑制细胞增殖、侵袭和迁移,促进其自噬和凋亡,并呈现剂量依赖性。

Objective To explore the effects of fangchinoline(FAN)on the biological behavior of prostate cancer PC3 cells based on mitochondrial apoptosis pathway.Methods Prostate cancer PC3 cells were divided into 4 groups according to FAN treatment concentrations,namely FAN 0μmol/L,5μmol/L,10μmol/L and 20μmol/L groups,which were treated with FAN 0μmol/L(DMSO replacement),5μmol/L,10μmol/L and 20μmol/L,respectively,then the experiment was carried out after 48 h incubation.The proliferation of PC3 cells was measured by CCK-8 method,the cloning ability of PC3 cells was determined by plate cloning method,the apoptosis of PC3 cells was detected by flow cytometry,the invasion of PC3 cells was checked by Transwell assay,the migration of PC3 cells was examined by scratch assay,and the autophagy related proteins and mitochondrial apoptosis pathway proteins of PC3 cells were measured by Western blot.Results Compared with FAN 0μmol/L group,the proliferation,clone formation, invasion and migration of PC3 cells in FAN 5 μmol/L, 10 μmol/L and 20 μmol/L groups were significantly lower (P<0.001), and the apoptosis rate was significantly higher (P<0.001). Compared with FAN 5 μmol/L and FAN 10 μmol/L groups, the proliferation, clone formation, invasion and migration of PC3 cells in FAN 20 μmol/L group were significantly lower (P<0.001). Compared with FAN 0 μmol/L group, the expressions of P62 and Bcl-2 in PC3 cells in FAN 5 μmol/L, 10 μmol/L and 20 μmol/L groups were significantly lower (P<0.001), and the expressions of LC3B, Bax and Caspase-3 proteins were significantly higher (P<0.001). Conclusion FAN may inhibit cell proliferation, invasion and migration through mitochondrial apoptosis pathway, and promote autophagy and apoptosis in a dose-de pendent manner.