钙敏感受体在3,3′-二吲哚甲烷抑制肝癌细胞增殖与迁移侵袭中的作用  

Effect of calcium-sensing receptor on the inhibition of proliferation,migration and invasion of hepatocellular carcinoma cell induced by 3,3′-diindolylmethane

作　　者：徐伟 杨奇[3] 陆定坤 肖钊 姜袁越 何大伟 孙俊 顾杰 叶洋 陆荣柱[1,5] XU Wei;YANG Qi;LU Dingkun;XIAO Zhao;JIANG Yuanyue;HE Dawei;SUN Jun;GU Jie;YE Yang;LU Rongzhu(School of Medicine,Jiangsu University,Zhenjiang Jiangsu 212013;Department of Pathology,Yancheng Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Traditional Chinese Medicine,Yancheng Jiangsu 224001;Department of Pathology,Zhenjiang Hospital of Chinese Traditional and Western Medicine,Zhenjiang Jiangsu 212000;Department of Pathology,Kunshan Hospital of Traditional Chinese Medicine,Suzhou Jiangsu 215300;Center for Experimental Research,the First People′s Hospital of Kunshan City,Suzhou Jiangsu 215300;Department of Hepatobiliary Surgery,the First People′s Hospital of Kunshan City,Suzhou Jiangsu 215300;School of Life Science,Jiangsu University,Zhenjiang Jiangsu 212013,China)

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]南京中医药大学附属盐城市中医院病理科,江苏盐城224001 [3]镇江市中西医结合医院病理科,江苏镇江212000 [4]昆山市中医医院病理科,江苏苏州215300 [5]昆山市第一人民医院临床实验研究中心,江苏苏州215300 [6]昆山市第一人民医院肝胆外科,江苏苏州215300 [7]江苏大学生命科学学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2022年第1期26-31,共6页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(82073543,81502800);江苏省自然科学基金青年基金项目(BK20140576);昆山市重点研发计划(社会发展)项目(KSF202139)。

摘　　要：目的:探讨钙敏感受体(calcium-sensing receptor,CaSR)在3,3′-二吲哚甲烷(3,3′-diindolylmethane,DIM)对肝癌细胞增殖、迁移与侵袭影响中的作用。方法:采用不同浓度(0、20、40、60、80μmol/L)DIM分别处理人肝癌HepG2和SMMC-7721细胞24 h,采用MTT法检测肝癌细胞增殖能力,蛋白免疫印迹法检测CaSR蛋白表达,筛选合适的DIM浓度,Transwell法检测DIM对肝癌细胞迁移和侵袭能力的影响。另将HepG2和SMMC-7721细胞分为4组:对照组、60μmol/L DIM组、300μmol/L氯化钆(CaSR激动剂)组及氯化钆+DIM组(用300μmol/L氯化钆预处理细胞0.5 h,再与60μmol/L DIM共处理24 h),评价细胞增殖、CaSR蛋白表达以及细胞迁移和侵袭能力。结果:与对照组相比,60μmol/L和40μmol/L DIM处理后可分别显著抑制HepG2和SMMC-7721细胞增殖能力(P均<0.05),降低两株细胞CaSR蛋白表达量(P均<0.05);DIM抑制两株细胞迁移和侵袭能力呈效应剂量关系;与60μmol/L DIM组相比,300μmol/L氯化钆+60μmol/L DIM组细胞CaSR蛋白表达、增殖、迁移和侵袭能力均明显升高(P<0.05)。结论:激活CaSR可以缓解DIM引起的抗肝癌效应,提示CaSR可能是DIM抗肝癌作用的潜在靶点。Objective:To investigate the role of calcium-sensing receptor(CaSR)in the effect of 3,3′-diindolylmethane(DIM)on the proliferation,migration and invasion of human hepatocellular carcinoma cells.Methods:Different concentrations of DIM(0,20,40,60 and 80μmol/L)were used to treat the human hepatocellularcarcinoma cells HepG2 and SMMC-7721 for 24 h.The proliferative ability was detected by MTT assay.Western blotting assay was used to detect the expression of CaSR.The optimal DIM concentration was selected based on the two results above.Transwell assay was performed to detect the ability of migration and invasion.The treatments of cells were divided into four groups:control group,60μmol/L DIM group,300μmol/L GdCl_(3)(CaSR agonist)group and 300μmol/L GdCl 3+60μmol/L DIM co-treatment group.The cells of last group were pretreated with 300μmol/L GdCl_(3) for 0.5 h,and then were co-treated with 60μmol/L DIM for 24 h.The above experiments were repeated to assess the proliferative ability,the expression of CaSR and the ability of migration and invasion of human hepatocellular carcinoma cells.Results:Compared with the control group,after treatment with 60μmol/L and 40μmol/L DIM,the proliferative ability of two cell lines was decreased significantly(P<0.05)and the expression of CaSR protein was also markedly decreased(P<0.05);DIM suppressed the ability of migration and invasion of the two cell lines in a concentration-dependent manner.Compared with 60μmol/L DIM group,the expression of CaSR,cell proliferation,migration and invasion ability were enhanced in the 300μmol/L GdCl 3+60μmol/L DIM co-treatment group(P<0.05).Conclusion:CaSR plays a role in anti-cancer effect by DIM and may be severed as a potential target for anti-liver cancer.

关 键 词：钙敏感受体 3 3′-二吲哚甲烷 肝癌 HEPG2 SMMC-7721 细胞增殖 细胞迁移与侵袭 

分 类 号：R285[医药卫生—中药学]