青藤碱对体外人胆管癌细胞的抑制作用

Inhibitory effects of sinomenine on cholangiocarcinoma cells in vitro

目的研究青藤碱对体外人胆管癌细胞的抑制作用。方法 CCK-8和平板克隆实验检测青藤碱对胆管癌细胞存活率及增殖率的影响,流式细胞术检测青藤碱对胆管癌细胞凋亡作用的影响,Transwell检测青藤碱对胆管癌细胞侵袭能力的影响,细胞划痕检测青藤碱对胆癌细胞迁移能力的影响,Western blot检测青藤碱对胆管癌细胞中凋亡相关蛋白caspase-3和caspase-9和侵袭相关蛋白E-Cadherin、Vimentin表达的影响。结果随着青藤碱浓度的提高以及时间的延长,胆管癌细胞的存活率逐渐下降(P<0.05,P<0.01),细胞集落形成数量逐渐减少(P<0.05,P<0.01)。青藤碱可呈剂量依赖性地促进胆管癌细胞凋亡(P<0.01),抑制胆管癌细胞的侵袭迁移(P<0.01),并且可以使cleaved caspase-3、cleaved caspase-9蛋白以及E-cadherin蛋白表达逐渐升高,vimentin蛋白表达逐渐下降(P<0.05,P<0.01)。结论青藤碱可以有效地抑制体外胆管癌细胞的增殖,侵袭和迁移能力,诱导细胞凋亡,这可能与其上调cleaved caspase-3、cleaved caspase-9和E-cadherin蛋白表达,以及下调vimentin蛋白表达有关。

AIM To investigate the inhibitory effects of sinomenine on cholangiocarcinoma cells in vitro. METHODS The changes of viability and proliferative rate of cholangiocarcinoma in response to sinomenine treatment were detected using CCK-8 assay and coloy formation assay. The flow cytometry was used to explore the effects of sinomenine on the cellular apoptosis of cholangiocarcinoma. Transwell assay and wound-healing assay were conducted to measure the cell invasion and migration abilities respectively. Western blot was applied to detecting the expression of apoptosis-related protein cleaved caspase-3 and cleaved caspase-9, as well as the invasion-related proteins E-cadherin and vimentin. RESULTS The viability and proliferative rate of cholangiocarcinoma cells were significantly inhibited by sinomenine in a time-dependent and dose-dependent manner(P<0.05, P<0.01),and the number of cell colony formation decreased gradually(P<0.05, P<0.01). In a dose-dependent manner, sinomenine increased cellular apoptosis(P<0.01), inhibited the cellular invasion and migration of cholangiocarcinoma(P<0.01), up-regulated the expressions of cleaved caspase-3,cleaved caspase-9 and E-cadherin, and down-regulated the expressions of vimentin(P<0.05, P<0.01).CONCLUSION Sinomenine can effectively inhibit the cellular proliferation,migration and invasion of human cholangiocarcinoma most likely via up-regulation of the protein levels of cleaved caspase-3,cleaved caspase-9 and E-cadherin,and down-regulation of the protein level of vimentin.