摘要
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.
作者
吕彦霖
朱昌毫
潘耀振
陈玲
喻超
孙诚谊
LüYan lin;ZHU Chang hao;PAN Yao zhen;CHEN Ling;YU Chao;SUN Cheng yi(Department of Hepatobiliary Surgery,Hospital Affiliated to Guizhou Medical University,Guiyang 550004,China)
出处
《中成药》
CAS
CSCD
北大核心
2019年第11期2601-2608,共8页
Chinese Traditional Patent Medicine
基金
贵州省肝胆外科临床医学研究中心(黔科合平台人才[2017]5404)
贵州省高层次创新人才培养计划“十”层次人才(黔科合平台人才[2016]5647)
贵州省科学技术厅-贵阳医学院院士工作站肝胆外科分站(黔科合院士站[2015]4013)
贵州省孙诚谊“肝胆胰脾疾病诊治”导师工作室(黔教研合GZS[2016]09)
贵州省科技厅贵州医科大学联合基金(黔科合[2015]7406)
关键词
青藤碱
胆管癌
增殖
凋亡
侵袭
转移
Sinomenine
cholangiocarcinoma
proliferation
apoptosis
invasion
migration