长链非编码RNALINC02587对前列腺癌细胞增殖和侵袭的影响及机制实验研究

Effect and mechanism of long non-coding RNA LINC02587 on the proliferation and invasion of prostate cancer cells

目的:探讨长链非编码RNA(lncRNA)LINC02587对前列腺癌细胞增殖和侵袭的影响及其分子机制。方法:采用LncRNADiseasev2.0数据库分析LINC02587在前列腺癌组织中的表达。RT-qPCR检测LINC02587在前列腺癌细胞系PC3、LNCaP、DU-145、22Rv1、C4-2B和正常前列腺上皮细胞RWPE-1中的表达水平,选取表达水平最高的细胞进行后续实验。将si-NC、si-LINC02587分别转染至22Rv1细胞,分为si-NC组和si-LINC02587组。采用CCK-8法检测22Rv1细胞增殖能力,Transwell实验检测22Rv1细胞侵袭能力,克隆形成实验检测22Rv1细胞克隆形成率。双荧光素酶报告基因实验验证LINC02587与miR-203的靶向关系。采用LncRNADiseasev2.0数据库分析LINC02587和miR-203在前列腺癌组织中表达的相关性。采用RT-qPCR检测两组22Rv1细胞miR-203表达水平。采用Westernblot检测高迁移率族ATHook蛋白2(HMGA2)、细胞周期蛋白依赖性蛋白激酶3(CDK3)、细胞周期蛋白C(CyclinC)、纤维连接蛋白(Fibronectin)以及sineoculis同源框同源物1(SIX1)蛋白表达。结果:与正常前列腺组织比较,LINC02587在前列腺癌组织中表达水平升高(P<0.01)。与RWPE-1细胞比较,PC3、LNCaP、DU-145、22Rv1以及C4-2B细胞中LINC02587表达水平升高,且22Rv1细胞表达水平最高(均P<0.01)。si-LINC02587组22Rv1细胞中LINC02587表达水平低于si-NC组(P<0.01)。与si-NC组比较,si-LINC02587组22Rv1细胞的增殖能力及克隆形成率降低、侵袭细胞数减少(均P<0.05)。LINC02587与miR-203存在靶向关系,LINC02587和miR-203在前列腺癌组织中的表达呈负相关(P<0.01)。与si-NC组比较,si-LINC02587组22Rv1细胞中miR-203表达水平升高,HMGA2、CDK3、CyclinC、Fibronectin、SIX1蛋白表达水平降低(均P<0.05)。结论:LINC02587在前列腺癌组织和细胞中高表达,下调LINC02587通过靶向miR-203抑制前列腺癌细胞的增殖和侵袭。

Objective:To explore the effect of lncRNA LINC02587 on the proliferation and invasion of prostate cancer cells and its molecular mechanism.Methods:The expression of LINC02587 in prostate cancer tissues was analyzed using the LncRNADisease v2.0 database.The expression levels of LINC02587 in human prostate cancer cell lines PC3,LNCaP,DU-145,22Rv1,C4-2B,and normal prostate epithelial cells RWPE-1 were detected by RT-qPCR,and the cell line with the highest expression was selected for subsequent experiments.si-NC and si-LINC02587 were transfected into 22Rv1 cells,which were then divided into si-NC group and si-LINC02587 group.CCK-8 assay was used to detect the proliferation of 22Rv1 cells,and Transwell assay was used to detect the invasion ability of 22Rv1 cells.Colony formation assay was used to detect the colony formation rate of 22Rv1 cells.Dual-luciferase reporter gene experiment was used to verify the targeting relationship between LINC02587 and miR-203.The LncRNADisease v2.0 database was used to analyze the correlation between LINC02587 and miR-203 expression in prostate cancer tissues.The expression levels of miR-203 in 22Rv1 cells in the two groups were detected by RT-qPCR.The protein expression of high mobility group AT-hook 2(HMGA2),cyclin-dependent kinase 3(CDK3),cyclin C,fibronectin and sine oculis homeobox homolog 1(SIX1)were detected by Western blot.Results:Compared with normal prostate tissues,the expression of LINC02587 in prostate cancer tissues was increased(P<0.01).Compared with RWPE-1 cells,the expression levels of LINC02587 in PC3,LNCaP,DU-145,22Rv1 and C4-2B cells were increased,with the highest expression in 22Rv1 cells(all P<0.01).The expression level of LINC02587 in si-LINC02587 group 22Rv1 cells was lower than that in the si-NC group(P<0.01).Compared with the si-NC group,the proliferation ability and colony formation rate of 22Rv1 cells in the si-LINC02587 group were decreased,and the number of invasive cells was reduced(all P<0.05).LINC02587 has a targeting relationship with miR-203,and the expression of LINC02587 and miR-203 in prostate cancer tissues is negatively correlated(P<0.01).Compared with the si-NC group,the expression level of miR-203 in the si-LINC02587 group 22Rv1 cells was increased,and the protein expression levels of HMGA2,CDK3,Cyclin C,Fibronectin and SIX1 were decreased(all P<0.05).Conclusion:LINC02587 is highly expressed in prostate cancer tissues and cells,and down-regulation of LINC02587 inhibits the proliferation and invasion of prostate cancer cells by targeting miR-203.