丫蕊花甾体皂苷YB16对人前列腺癌细胞PC-3增殖与凋亡的影响

Effect of Steroidal Saponin YB16 of Ypsilandra Thibetica on Proliferation and Apoptosis of Human Prostate Cancer Cells PC-3

摘要:

目的：探讨丫蕊花甾体皂苷YB16对人前列腺癌细胞PC-3增殖与凋亡的影响，并探讨其作用机制。方法：体外培养PC-3细胞，给予不同浓度的YB16（0.125~16 μmol·L^-1），以噻唑蓝（MTT）比色法检测YB16对PC-3的细胞毒性，倒置相差显微镜观察细胞形态变化，吖啶橙（AO）染色、流式细胞术检测YB16对PC-3的凋亡影响，逆转录-聚合酶链式反应（RT-PCR）检测YB16对PC-3细胞B淋巴细胞瘤-2（Bcl-2），B细胞淋巴瘤-xl（Bcl-xl），Bcl-2相关X蛋白（Bax），半胱氨酸蛋白酶-3（Caspase-3）mRNA的表达；蛋白免疫印迹法（Western blot）检测Bcl-2，Bcl-xl，Bax，激活型Caspase-3（cleaved-Caspase-3）蛋白表达，并对结果进行分析。结果：YB16能显著抑制PC-3细胞的生长，具有剂量依赖性（P<0.05）；YB16能促进细胞的凋亡，相差显微镜，AO染色法观察可见细胞具凋亡特征性改变；YB16能下调Bcl-2，Bcl-xl，上调Bax，Caspase-3 mRNA和蛋白的表达（P<0.05）。结论：YB16能抑制PC-3细胞增殖，促进PC-3发生凋亡，其机制可能与促进Caspase-3表达有关，具有良好的抗肿瘤作用。

Abstract:

Objective: To investigate the effects of Ypsilandra thibetica 16 (YB16) extracted from the Ypsilandra thibetica on proliferation and apoptosis of human prostate cancer cells PC-3, and explore its mechanism. Method: PC-3 cells were cultured in vitro and treated with different concentrations of YB16 (0.125~16 μmol·L^-1). The effects of YB16 on PC-3 cytotoxicity were detected by (4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, and the cell morphology was observed by phase contrast microscope. Acridine orange (AO) staining and flow cytometry were used to detect the apoptosis of PC-3 cells induced by YB16.Reverse-transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression levels of B-cell lymphoma 2 gene (Bcl-2), B-cell lymphoma 2 gene-xl (Bcl-xl), Bcl-2 associated X protein (Bax), and cysteinyl aspartate specific proteinase-3(Caspase-3)of PC-3 cells, and Western blot was used to detect the protein expression levels of Bcl-2, Bcl-xl, Bax and cleaved-Caspase-3. Result: YB16 could significantly inhibit the growth of PC-3 cells in a dose-dependent manner (P<0.05). YB16 could promote PC-3 cells apoptosis, and PC-3 cells had obvious characteristic apoptosis changes in the observation under phase contrast microscope and AO staining. YB16 could reduce the mRNA and protein expression levels of Bcl-2, Bcl-xl, but increase the mRNA and protein expression levels of Bax, Caspase-3 (P<0.05). Conclusion: YB16 can inhibit PC-3 cells proliferation, and induce apoptosis obviously. YB16 has a potential antitumor value, and its mechanism may be associated with promoting Caspase-3 expression.