microRNA-384靶向HDAC9调控肝癌细胞增殖迁移侵袭和凋亡的机制研究  

Molecular mechanism of microRNA-384 regulating the biological characteristics of proliferation, migration, invasion and apoptosis of hepatic carcinoma cells by targeting HDAC9

作　　者：刘皎皎 杨跃青 戴玲 王佩 叶苗清 李煜国 LIU Jiao-jiao;YANG Yue-qing;DAI Ling;WANG Pei;YE Miao-qing;LI Yu-guo(Department of Hepatic,Shaanxi Traditional Chinese Medicine Hospital,Xi’an 710003,China;Departmentof Hepatic,Hubei Traditional Chinese Medicine Hospital,Wuhan 430060,China)

机构地区：[1]陕西省中医医院肝病科,陕西西安710003 [2]湖北省中医院光谷院区肝病科,湖北武汉430060

出　　处：《中国中西医结合消化杂志》2020年第8期573-579,共7页Chinese Journal of Integrated Traditional and Western Medicine on Digestion

基　　金：国家自然科学基金(No:81373513);国家中医临床研究基地(湖北)重点病种研究项目(No:JDZX2012054,JDZX2015172)。

摘　　要：[目的]:研究microRNA-384(miR-384)、人组蛋白去乙酰化酶(HDAC9)对肝癌细胞增殖、迁移侵袭及凋亡的影响,并探讨其作用机制。[方法]运用qRT-PCR检测正常肝上皮细胞THLE3和肝癌细胞HEP3B、BEL-7402、HUH7中HDAC9、miR-384的mRNA的表达;将PC DNA组(转染PC DNA)、PC DNA-HDAC9组(转染PC DNA-HDAC9)、anti-miR-NC组(转染anti-miR-NC)、anti-miR-384组(转染anti-miR-384)、si-NC组(转染si-NC)、si-HDAC9组(转染si-HDAC9)、PC DNA-HDAC9+miR-NC组(共转染PC DNA-HDAC9和miR-NC)、PC DNA-HDAC9+miR-384组(共转染PC DNA-HDAC9和miR-384),用脂质体法转染至HUH7;Western blot检测细胞中HDAC9的蛋白表达;Transwell小室检测细胞迁移和侵袭;流式细胞术检测细胞凋亡;双荧光素酶报告基因实验检测细胞的荧光素酶活性。[结果]与THLE3细胞比较,肝癌细胞HEP3B、BEL-7402、HUH7中HDAC9的mRNA和蛋白均显著上调,miR-384的mRNA显著下调(P<0.05);过表达HDAC9可明显促进HUH7的增殖、迁移侵袭,抑制凋亡,敲减HDAC9具有相反的结果;HDAC9是miR-384的靶标,且受miR-384的负向调控。过表达miR-384可逆转HDAC9对肝癌细胞的作用。[结论]miR-384抑制肝癌细胞增殖、迁移和侵袭,促进凋亡,其机制与靶向负调控HDAC9相关,将可为肝癌的治疗提供新的治疗靶点。[Objective]To study the effects of HDAC9 and microRNA-384(miR-384) on proliferation,migration,invasion and apoptosis of hepatic carcinoma cells,and to explore its mechanism.[Methods]qRT-PCR was used to detect the expression of HDAC9 and miR-384 mRNA in normal liver epithelial cells THLE3 and hepatic carcinoma cells Hep3 B,BEL-7402 and HUH7;pc DNA group(transfected pc DNA),pc DNA-HDAC9 group(transfected pc DNA-HDAC9),anti-miR-NC group(transfected anti-miR-NC),anti-miR-384 group(transfected anti-miR-384),si-NC group(transfected si-NC),si-HDAC9 group(transfected si-HDAC9),pc DNA-HDAC9+miR-NC group(co-transfected pc DNA-HDAC9 and miR-NC)and pc DNA-HDAC9+miR-384 group(co-transformed PC DNA-HDAC9 and miR-384)were transfected into HUH7 by liposome method;protein expression of HDAC9 in cells was detected by Western Blot;cell migration and invasion were detected by Transwell Chamber;apoptosis was detected by flow cytometry;the prime enzyme reporter assay detects the luciferase activity of the cells.[Results]Compared with THLE3,the mRNA and protein of HDAC9 in Hep3 B,BEL-7402 and HUH7 were significantly up-regulated,and the mRNA of miR-384 was significantly down-regulated(P<0.05).Overexpression of HDAC9 significantly promoted proliferation,migration and inhibition of apoptosis in HUH7 while knockdown of HDAC9 has the opposite effect;HDAC9 is targeted and negatively regulated by miR-384.Overexpression of miR-384 reverses the regulation of HDAC9 on hepatoma cells.[Conclusion]miR-384 inhibits the proliferation,migration,invasion and apoptosis of hepatocellular carcinoma cells,and its mechanism is related to the targeting negative regulation of HDAC9,providing a new therapeutic target for the treatment of hepatic carcinoma.

关 键 词：miR-384 HDAC9 肝癌 增殖 迁移 侵袭 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]