miR-129-3p与LPAR3在肝癌细胞中的表达水平及其靶向关系研究  

Expression level and targeting relationship of miR-129-3p and LPAR3 in hepatoma cells

作　　者：贾明君 王义真 常聪颖 张银阁 JIA Mingjun;WANG Yizhen;CHANG Congying;ZHANG Yinge(No.1 Department of Hepatology,Xingtai Second Hospital,Xingtai,Hebei 054000,China;Department of Hepatobiliary Surgery,Xingtai Third Hospital,Xingtai,Hebei 054000,China)

机构地区：[1]邢台市第二医院肝病一科,河北邢台054000 [2]邢台市第三医院肝胆外科,河北邢台054000

出　　处：《国际检验医学杂志》2023年第1期69-73,共5页International Journal of Laboratory Medicine

基　　金：邢台市重点研发计划自筹项目(2021ZC090)。

摘　　要：目的探究微小RNA(miR)-129-3p与溶血磷脂酸受体3(LPAR3)在肝癌细胞中的表达水平及其靶向调控机制。方法细胞学实验:选取3种肝癌细胞系HepG2、BEL-7402、SMMC-7721及正常肝细胞系HL-7702作为研究对象,通过实时荧光定量聚合酶链式反应(qPCR)检测miR-129-3p与LPAR3的表达水平,生物信息学软件DBmiR预测miR-129-3p与LPAR3的靶向关系,并使用双荧光报告实验探究miR-129-3p与LPAR3在4种细胞系HepG2、BEL-7402、SMMC-7721和HL-7702的靶向关系。裸鼠荷瘤实验:将20只裸鼠分为野生组(n=10)和miR-129-3p组(n=10),分别接种野生型HepG2细胞和转染miR-129-3p的HepG2细胞,绘制肿瘤生长曲线并通过小动物活体成像检测肿瘤体积,qPCR和western blot检测肿瘤组织miR-129-3p,LPAR3的表达水平及PI3K/AKT信号通路分子。结果细胞实验表明,与对照细胞相比,3种肝癌细胞中miR-129-3p显著低表达,LPAR3显著高表达,差异有统计学意义(P<0.01),生物信息学软件和双荧光报告实验证实miR-129-3p与LPAR3具有靶向调控关系(P<0.01)。裸鼠荷瘤实验表明:与对照组相比,miR-129-3p组肿瘤生长显著受到抑制,肿瘤体积显著降低,qPCR和Western blot实验表明肿瘤组织中miR-129-3p显著升高,LPAR3、PI3K、AKT表达水平显著降低,差异有统计学意义(P<0.01)。结论miR-129-3p在肝癌细胞中显著低表达,与LPAR3具有靶向调控关系;miR-129-3p的高表达水平可以显著抑制肿瘤生长,其机制可能与PI3K/AKT信号通路的调控相关。Objective To investigate the expression level and targeted regulation mechanism of microRNA(miR)-129-3p and lysophosphatidic acid receptor 3(LPAR3)in hepatoma cells.Methods Cytology experiment:three liver cancer cell lines HepG2,BEL-7402,SMMC-7721 and normal liver cell line HL-7702 were selected as the research objects.The expression levels of miR-129-3p and LPAR3 were detected by qPCR,the targeting relationship between miR-129-3p and LPAR3 was predicted by bioinformatics software DBmiR and double fluorescence report experiment was used to explore the targeting relationship between miR-129-3p and LPAR3 in four cell lines HepG2,BEL-7402,SMMC-7721 and HL-7702.Nude mice tumor bearing experiment:20 nude mice were divided into two groups,wild group(n=10)and miR-129-3p group(n=10).The two groups were inoculated with wild type HepG2 cells and HepG2 cells transfected with miR-129-3p respectively.The tumor growth curve was drawn and the tumor volume was detected by small animal imaging in vivo.The expression levels of miR-129-3p,LPAR3 and PI3K/AKT signal pathway molecules in tumor tissues were detected by qPCR and Western blot.Results Compared with the control cells,the expression of miR-129-3p was significantly lower and LPAR3 was significantly higher in the three hepatoma cells(P<0.01).Bioinformatics software and double fluorescence report experiment confirmed that miR-129-3p had a targeted regulatory relationship with LPAR3(P<0.01).The tumor bearing experiment in nude mice showed that compared with the control group,the tumor growth of miR-129-3p group was significantly inhibited and the tumor volume was significantly reduced.qPCR and Western blot showed that the expression levels of miR-129-3p,LPAR3,PI3K and AKT in tumor tissues were significantly increasedand,the differences were statistically decreasd(P<0.01).Conclusion The expression of miR-129-3p is significantly low in hepatoma cells and has a targeted regulatory relationship with LPAR3.The overexpression of miR-129-3p could significantly inhibit tumor growth,whi

关 键 词：微小RNA-129-3p 肝癌 溶血磷脂酸受体3 靶向调控 PI3K/AKT信号通路 

分 类 号：R735.7[医药卫生—肿瘤]