毛茛总苷通过circ_0079593/miR-324-5p通路抑制肝癌细胞增殖、迁移及侵袭的机制研究  

Study on the mechanism of total glycosides of ranunculus japonicus inhibiting the proliferation,migration and invasion of liver cancer cells through the circ_0079593/miR-324-5p pathway

作　　者：杜红蕾 张锋[1] 张咏华[1] 班志超 王旭 Du Honglei;Zhang Feng;Zhang Yonghua;Ban Zhichao;Wang Xu(Baoding No.2 Central Hospital,Zhuozhou 072750,Hebei,China)

机构地区：[1]保定市第二中心医院,河北涿州072750

出　　处：《肝癌电子杂志》2022年第1期48-54,共7页Electronic Journal of Liver Tumor

摘　　要：目的:探讨毛茛总苷对肝癌细胞增殖、迁移、侵袭的影响及其对circ_0079593/miR-324-5p通路的调控作用。方法:体外培养人肝癌细胞MHCC97H,并分为对照组,毛茛总苷低、中、高剂量组,si-NC组,si-circ_0079593组,毛茛总苷+pcDNA组,毛茛总苷+pcDNA-circ_0079593组;采用CCK-8法、Transwell实验分别检测细胞活力、迁移及侵袭;采用定量反转录聚合酶链反应(quantitative reverse transcription polymerase chain reaction,qRT-PCR)检测circ_0079593和miR-324-5p的表达水平;采用双荧光素酶报告基因实验检测circ_0079593和miR-324-5p的靶向关系;采用蛋白质印迹法检测基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的表达水平。结果:与对照组比较,毛茛总苷不同剂量组细胞增殖抑制率和miR-324-5p表达水平均升高(P<0.05),迁移及侵袭细胞数均减少(P<0.05),circ_0079593表达水平及MMP-2、MMP-9蛋白水平均降低(P<0.05),且呈剂量依赖性;circ_0079593能靶向调控miR-324-5p;与si-NC组比较,si-circ_0079593组细胞增殖抑制率升高(P<0.05),迁移及侵袭细胞数均减少(P<0.05),MMP-2、MMP-9蛋白水平均降低(P<0.05);与毛茛总苷+pcDNA组比较,毛茛总苷+pcDNA-circ_0079593组细胞增殖抑制率降低(P<0.05),迁移及侵袭细胞数均增多(P<0.05),MMP-2、MMP-9蛋白水平均升高(P<0.05)。结论:毛茛总苷可通过调控circ_0079593/miR-324-5p通路降低肝癌细胞增殖、迁移及侵袭能力。Objective:To explore the effect of total glycosides of ranunculus japonicus on the proliferation,migration and invasion of liver cancer MHCC97H cells and its regulation of circ_0079593/miR-324-5p pathway.Method:Human hepatocarcinoma cells MHCC97H were cultured in vitro,experimental groups:control group,total glycosides of ranunculus japonicus low-dose group,total glycosides of ranunculus japonicus medium-dose group,total glycosides of ranunculus japonicus high-dose group,si-NC group,si-circ_0079593 group,total glycosides of ranunculus japonicus+pcDNA group,total glycosides of ranunculus japonicus+pcDNA-circ_0079593 group.CCK-8 and Transwell chamber experiments were used to detect cell viability,migration and invasion.The quantitative reverse transcription polymerase chain reaction(qRT-PCR)method was used to detect the expression of circ_0079593 and miR-324-5p.The dual luciferase reporter gene experiment was used to detect the targeting relationship between circ_0079593 and miR-324-5p.Western blotting was used to detect the protein expression of matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase-9(MMP-9).Result:Compared with the control group,the cell proliferation inhibition rate and the expression level of miR-324-5p in different dose groups of total glycosides of ranunculus japonicus were increased(P<0.05),while the number of migrating and invading cells was decreased(P<0.05),and the expression level of circ_0079593 and the protein levels of MMP-2,MMP-9 were decreased(P<0.05),and were dose-dependent.circ_0079593 could target and regulate miR 324-5p.Compared with the si-NC group,the cell proliferation inhibition rate in the si-circ_0079593 group was increased(P<0.05),while the number of migrating and invading cells was decreased(P<0.05),and the protein levels of MMP-2 and MMP-9 were decreased(P<0.05).Compared with the total ranunculus glycosides+pcDNA group,the inhibition rate of cell proliferation in the total ranunculus glycosides+pcDNA-circ_0079593 group was reduced(P<0.05),while the number of migr

关 键 词：毛茛总苷 肝癌 细胞增殖 细胞迁移 细胞侵袭 

分 类 号：R735.7[医药卫生—肿瘤]