摘要
目的研究钙离子通道抑制剂逆转肝癌细胞多药耐药和对异黏蛋白表达的影响及分子机制。方法采用不同浓度钙离子抑制剂(0、25、50、100、200、400μmol/L)干预肝癌MHCC97H、7402细胞不同时间(12、24、48 h)后,用体外细胞划痕实验检测处理后肝癌细胞运动迁徙能力;采用RT-PCR和免疫细胞化学技术检测钙离子抑制剂对肝癌细胞中P-gp和异黏蛋白的表达的影响。结果钙离子抑制剂能够一过性抑制肝癌MHCC97H、7402细胞的迁徙运动能力,且具有一定的时间和浓度依赖性特点(P<0.05)。同时,不同浓度钙离子抑制剂盐酸地尔硫卓均能一过性上调肝癌MHCC97H、7402细胞中异黏蛋白mRNA和蛋白的表达(P<0.05),但并不抑制P-gp蛋白的表达。结论钙离子抑制剂地尔硫卓能一过性抑制MHCC97H、7402肝癌细胞的迁徙运动,并能反馈性上调促癌基因异黏蛋白mRNA和蛋白的表达水平。因而,采用钙离子抑制剂很可能具有增加肿瘤进展的潜在风险。
Objective To investigate the effect of calcium channel blocker diltizem in reversing multi-drug resistance(MDR) and on metadherin expression in hepatocellular carcinoma cells and explore the molecular mechanism. Methods Hepatocellular carcinoma MHCC97 H and 7402 cells were treated with diltiazem hydrochloride, a calcium channel blocker(0, 25, 50, 100, 200,and 400 μmol/L), for 12, 24, or 48 h. Wound healing assay was employed to assess the changes in the mobility and migration of the cells following the treatments, and the changes in the expression levels of metadherin mRNA and protein and P-gp protein were determined using RT-PCR and immunocytochemistry. Results Diltiazem hydrochloride could transiently inhibit the migration and movement of MHCC97 H and 7402 cells in vitro in a time-and concentration-dependent manner(P<0.05).Diltiazem hydrochloride at different concentrations also transiently up-regulated the expressions of metadherin mRNA and protein but did not inhibit the expression of P-gp protein in MHCC97 H and 7402 cells. Conclusion Calcium channel blocker can transiently inhibit the migration of hepatocellular carcinoma cells in vitro and up-regulate the expression of metadherin mRNA and protein through a feedback mechanism, suggesting the potential risk of calcium channel blockers for promoting tumor progression during the treatment of malignant tumors.
作者
郭睿
金雪媛
田怡
黄小钟
李宗芳
杨军
GUO Rui;JIN Xueyuan;TIAN Yi;HUANG Xiaozhong;LI Zongfang;YANG Jun(Department of Pathology,Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710004,China;National&Local Joint Engineering Research Center of Biodiagnostics and Biotherapy,Xi'an 710004,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2019年第3期298-303,共6页
Journal of Southern Medical University
基金
国家自然科学基金(NSFC30872403)
教育部新世纪优秀人才计划(NCET-10-0647)~~
关键词
肝细胞癌
多药耐药
钙离子抑制剂
地尔硫卓
异黏蛋白
hepatocellular carcinoma
multi-drug resistance
diltizem
metadherin
calcium channel blockers
