派特灵对HeLa细胞增殖迁移能力及PI3K/Akt信号转导通路的影响  

Effect of Paiteling on HeLa Cell Proliferation and Metastasis Ability and PI3K/Akt Signal Transduction Pathway

作　　者：刘运华[1] 赵宗江[1] 张新雪[1] 杨涛[1] 张萌萌 吴英杰[1] 高坤[1] 郑鹏飞 LIU Yun-hua;ZHAO Zong-jiang;ZHANG Xin-xue;YANG Tao;ZHANG Meng-meng;WU Ying-jie;GAO Kun;ZHENG Peng-fei(Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学,北京100029

出　　处：《中国实验方剂学杂志》2020年第17期56-63,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：派特灵合作开发项目(2180071720049)。

摘　　要：目的:探索派特灵对HeLa细胞增殖迁移侵袭能力及磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)信号通路相关蛋白的影响。方法:①将HeLa细胞分为空白组,派特灵组(3.906,2.604,1.953,1.563,1.302,1.116,0.977 g·L^-1),加药干预24 h,镜下观察细胞形态变化,并采用噻唑蓝(MTT)比色法检测细胞活力,计算派特灵对HeLa细胞的半数抑制浓度(IC50)。②将HeLa细胞分为空白组,顺铂组(0.01 g·L^-1),派特灵高、中、低质量浓度组(2.974,1.487,0.991 g·-1),采用细胞增殖与毒性检测(CCK-8)法检测派特灵对HeLa细胞增殖能力的影响,使用划痕实验检测细胞的迁移情况,采用侵袭实验(Transwell)检测细胞侵袭能力的变化。③增设抑制剂LY294002组(0.006 g·L^-1),采用蛋白免疫印迹法(Western blot)检测派特灵对PI3K,Akt,重组人B细胞淋巴瘤因子-xl(Bcl-xl),B细胞淋巴瘤/白血病相关d蛋白(Bad)表达的影响。结果:①与空白组比较,镜下观察显示,用药组细胞数目减少,细胞形态不完整;MTT比色法显示,派特灵对HeLa细胞增殖有显著抑制作用(P<0.01),计算得出派特灵对HeLa细胞的IC50为2.974 g·L^-1。②CCK-8法显示,与空白组比较,培养24,36,48 h,所有用药组均对HeLa细胞增殖有抑制作用(P<0.01);与顺铂组比较,各时间点派特灵中、低质量浓度组对HeLa细胞增殖抑制作用减弱(P<0.01);划痕实验显示,与空白组比较,各加药组均能抑制HeLa细胞的迁移能力(P<0.01),派特灵高质量浓度组细胞迁移率低于顺铂组(P<0.05);Transwell实验显示,与空白组比较,各加药组HeLa细胞穿膜数减少(P<0.01),与顺铂组比较,派特灵中、低质量浓度组细胞穿膜数升高(P<0.01)。③Western blot显示,与空白组比较,派特灵高、中、低剂量及LY294002组的PI3K,Bcl-xl,Akt表达量下降(P<0.05,P<0.01),Bad表达量增高(P<0.01);与派特灵高质量浓度组比较,派特灵低质量浓度组的PI3K,Akt,Bcl-xl蛋白表达升高(P<0.01),Bad表达降低(P<0.01)。结论:派�Objective:To explore the effect of Paiteling on the proliferation,metastasis and invasion of HeLa cells and relevant proteins of phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway.Method:①HeLa cells were divided into blank group and Paiteling concentration gradient groups(3.906,2.604,1.953,1.563,1.302,1.116,0.977 g·L^-1).After drug intervention for 24 h,the cell morphological changes were observed under microscope.The cell viability was measured by thiazole blue(MTT)colorimetry,and the half inhibitory concentration(IC50)of Paiteling on HeLa cells was calculated.②HeLa cells were divided into blank group,cisplatin group(0.01 g·L^-1),Paiteling high-dose group(2.974 g·L^-1),Paiteling medium-dose group(1.487 g·L^-1)and Paiteling low-dose group(0.991 g·L^-1).Cell proliferation and toxicity test(CCK-8)method was used to detect the effect of Paiteling on the proliferation ability of HeLa cells,scratch test was used to detect cell migration,and invasion test(Transwell)was used to detect changes in cell invasion ability.③Inhibitor LY294002 group(0.006 g·L^-1)was added.Western blot(WB)was used to detect the expressions of Paiteling on PI3K,Akt,recombinant human B-cell lymphoma factor-xl(Bcl-xl),and B-cell lymphoma/leukemia associated D protein(Bad).Result:①Compared with the blank group,microscopic observation showed that the number of cells in the treatment group was significantly reduced,and the cell morphology was incomplete.MTT experiments showed that Paiteling has a significantly inhibitory effect on HeLa cell proliferation(P<0.01).The IC50 of Paiteling on HeLa cells was calculated as 2.974 g·L^-1.②The CCK-8 experiment showed that compared with the blank group,all the drug-treated groups had an inhibitory effect on HeLa cell proliferation at 24,36,48 h(P<0.01),compared with the cisplatin group,middle and low-dose Paiteling groups showed a reduced inhibitory effect on HeLa cell proliferation at each time point(P<0.01).The scratch test showed that,compared with the blank group,each

关 键 词：HELA细胞 派特灵 半数抑制浓度(IC50) 侵袭 转移 磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt) 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]