欧前胡素抑制人肾癌786-O细胞增殖并促进其凋亡  

Imperatorin inhibits proliferation and promotes apoptosis of human renal carcinoma cell line 786-O

作　　者：景红梅 樊海燕[3] 黄小勇[1] 杜娟[1] 张静[1] 慕明涛[1] 史海燕[1] JING Hongmei;FAN Haiyan;HUANG Xiaoyong;DU Juan;ZHANG Jing;MU Mingtao;SHI Haiyan(Medical Research and Experimental Center,Medical College,Yan’an University,Yan’an 716000,China;Department of Pediatrics,Yan’an Baota District People’s Hospital;Department of Laboratory,First Hospital of Yulin)

机构地区：[1]延安大学医学院医学研究实验中心,延安716000 [2]延安市宝塔区人民医院儿科 [3]榆林市第一医院检验科

出　　处：《山西医科大学学报》2023年第10期1323-1330,共8页Journal of Shanxi Medical University

基　　金：延安市重点研发计划项目(SL2022SLSFGG-116)。

摘　　要：目的探究欧前胡素(imperatorin)对人肾癌细胞786-O增殖及凋亡的影响,初步研究其作用机制。方法分别使用0,20,40,60,80,100,120,140,160,180,200μmol/L的欧前胡素处理786-O细胞24,48,72 h后,CCK-8法检测786-O细胞增殖,并计算IC 50,筛选最佳干预浓度进行后续实验。生物显微镜观察细胞形态变化;CCK-8实验和克隆形成实验检测各组细胞增殖能力的变化;流式细胞术检测各组细胞凋亡的情况;RT-qPCR和Western blot方法检测各组Fas、Caspase-8、Caspase-3、Bcl-2的表达情况。从TCMSP和HERB数据库收集欧前胡素作用靶点。由GeneCards数据库获取肾癌疾病靶点,使用STRING数据库构建蛋白互作网络,并筛选欧前胡素治疗肾癌的核心靶点。结果欧前胡素作用786-O细胞24 h和48 h的IC 50分别为94.36μmol/和65.66μmol/L。与0μmol/L组相比,40μmol/L组786-O细胞体积减小,细胞密度略减少,增殖能力受到抑制(P<0.01),细胞凋亡率增加(P<0.01);80μmol/L组细胞壁皱缩明显,细胞密度明显减少,增殖能力明显被抑制(P<0.001),细胞凋亡率明显增加(P<0.001)。RT-qPCR和Western blot实验结果显示,与0μmol/L组相比,40μmol/L组Fas、Caspase-8、Caspase-3的表达上调(P<0.05),80μmol/L组上述3个分子表达明显上调(P<0.01),Bcl-2则无明显变化。通过网络药理学方法筛选出7个欧前胡素治疗肾癌的核心靶点,分别为GSK3β、CDK2、PPARG、ESR1、MAPK14、AR、PTGS2。结论欧前胡素可能是通过靶向GSK3β调控死亡受体途径,进而抑制肾癌786-O细胞的增殖,并促进其凋亡。Objective To investigate the effect of imperatorin on the proliferation and the apoptosis of human renal cell carcinoma 786-O and explore its mechanism.Methods 786-O cells were treated with 0,20,40,60,80,100,120,140,160,180,200μmol/L imperatorin for 24,48,72 h,respectively.The cell proliferation activity of 786-O was determined by CCK-8 method,and IC 50 was calculated to screen the best intervention concentration for subsequent experiments.The morphological changes of 786-O cells were observed under a biological microscope.The cell proliferation ability was detected by CCK-8 assay and clone formation assay.Apoptosis was detected by flow cytometry.The expressions of Fas,Caspase-8,Caspase-3 and Bcl-2 were detected by RT-qPCR and Western blot.The targets of imperatorin were collected from TCMSP and HERB databases,the disease targets of renal cancer were obtained from GeneCards database,the protein interaction network was constructed using STRING database,and the core targets of imperatorin in the treatment of renal cancer were screened.Results The IC 50 of imperatorin on 786-O cells for 24 h and 48 h was 94.36μmol/L and 65.66μmol/L,respectively.Compared with 0μmol/L group,the volume of 786-O cells was decreased in 40μmol/L group,the cell density was slightly decreased,the proliferation ability was inhibited(P<0.01),and the apoptosis rate increased(P<0.01).Compared with 0μmol/L group,the cell wall was significantly shrunk in 80μmol/L group,the cell density was obviously reduced,the proliferation ability was significantly inhibited(P<0.001),and the apoptosis rate was significantly increased(P<0.001).The results of RT-qPCR and Western blot showed that compared with 0μmol/L group,the expressions of Fas,Caspase-8,and Caspase-3 were upregulated in 40μmol/L group(P<0.05),and the expressions of the above three molecules were significantly upregulated in 80μmol/L group(P<0.01),but Bcl-2 expression showed no significant change.Seven key targets of imperatorin in the treatment of renal cell carcinoma were screened by

关 键 词：欧前胡素 肾癌 细胞增殖 细胞凋亡 死亡受体途径 网络药理学 

分 类 号：R737.11[医药卫生—肿瘤]