LINC00355调控miR-494-3p/CCND2对前列腺癌细胞增殖、侵袭、迁移的影响  

Effects of LINC00355 on regulation of miR-494-3p/CCND2 on proliferation, invasion and migration of prostate cancer cells

作　　者：张建育[1] 李毅宁[1] 郭一泓[1] 穆鑫[1] ZHANG Jian-Yu;LI Yi-Ning;GUO Yi-Hong;MU Xin(Department of Urology Surgery,the Second Affiliated Hospital of Quanzhou Medical University,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属第二医院泌尿外科,泉州362000

出　　处：《中国免疫学杂志》2021年第5期564-570,共7页Chinese Journal of Immunology

基　　金：福建省自然科学基金项目(No.2018J01282)。

摘　　要：目的:探讨长链非编码RNA(LncRNA LINC00355)对前列腺癌细胞增殖、侵袭、迁移的影响及其作用机制。方法:qRT-PCR检测前列腺癌组织及癌旁组织中LINC00355、miR-494-3p、细胞周期蛋白HD2(CCND2)mRNA的表达水平;体外培养前列腺癌细胞DU145,分别将si-NC、si-LINC00355、si-LINC00355与anti-miR-NC、si-LINC00355与anti-miR-494-3p、si-LINC00355与miR-NC、si-LINC00355与miR-494-3p mimics共转染至DU145细胞。MTT检测细胞增殖;Transwell小室实验检测细胞迁移及侵袭能力;双荧光素酶报告实验验证LINC00355与miR-494-3p的靶向调控作用以及miR-494-3p与CCND2的靶向调控作用;Western blot检测CCND2、P21、上皮钙黏附素(E-cadherin)、基质金属蛋白酶-2(MMP-2)蛋白表达量。结果:与癌旁组织相比,前列腺癌组织中LINC00355与CCND2 mRNA的表达水平显著升高(P<0.05),miR-494-3p的表达水平显著降低(P<0.05);与si-NC组、si-LINC00355+miR-NC组相比,si-LINC00355组、si-LINC00355+miR-494-3p组细胞增殖抑制率显著升高(P<0.05),迁移细胞数与侵袭细胞数显著减少(P<0.05),P21、E-cadherin蛋白水平显著升高(P<0.05),MMP-2蛋白水平显著降低(P<0.05);与si-NC组相比,si-LINC00355组细胞中miR-494-3p的表达水平显著升高(P<0.05),CCND2 mRNA及蛋白表达水平均显著降低(P<0.05);与si-LINC00355+anti-miR-NC组相比,si-LINC00355+anti-miR-494-3p组细胞增殖抑制率显著降低(P<0.05),迁移细胞数与侵袭细胞数显著增多(P<0.05),P21、E-cadherin蛋白水平显著降低(P<0.05),MMP-2蛋白水平显著升高(P<0.05);双荧光素酶报告实验证实LINC00355能够靶向结合miR-494-3p, miR-494-3p能够靶向CCND2。结论:LINC00355通过与CCND2竞争结合miR-494-3p,降低miR-494-3p对CCND2表达的抑制作用,从而促进前列腺癌细胞增殖、侵袭、迁移。Objective:To investigate the effect of LncRNA LINC00355 on the proliferation, invasion and migration of prostate cancer cells and its mechanism of action.Methods:qRT-PCR was used to detect the expression levels of LINC00355,miR-494-3 p and CCND2 mRNA in prostate cancer tissues and adjacent tissues.Prostate cancer cell DU145 was cultured in vitro.si-NC,si-LINC00355,si-LINC00355 and anti-miR-NC,si-LINC00355 and anti-miR-494-3 p, si-LINC00355 and miR-NC,si-LINC00355 co-transfected with miR-494-3 p mimics into DU145 cells.MTT is used to detect cell proliferation.Transwell chamber experiments were performed to detect cell migration and invasion.The double luciferase report experiment verified the targeted regulation of LINC00355 and miR-494-3 p, and the targeted regulation of miR-494-3 p and CCND2.Western blot was used to detect the expression of CCND2,P21,E-cadherin and MMP-2 proteins.Results:Compared with adjacent cancer tissues, the expression levels of LINC00355 and CCND2 mRNA in prostate cancer tissues were significantly increased(P<0.05),and the expression levels of miR-494-3 p were significantly decreased(P<0.05).Compared with the si-NC group and si-LINC00355+ miR-NC group, the si-LINC00355 group, si-LINC00355+ miR-494-3 p group had significantly higher cell proliferation inhibition rates(P<0.05),the number of migrating cells and the number of invasion cells were significantly reduced(P<0.05),the levels of P21 and E-cadherin proteins were significantly increased(P<0.05),and the levels of MMP-2 protein were significantly reduced(P<0.05).Compared with the si-NC group, the expression level of miR-494-3 p in the si-LINC00355 group was significantly increased(P<0.05),and the levels of CCND2 mRNA and protein were significantly reduced(P<0.05).Compared with the si-LINC00355+ anti-miR-NC group, the cell proliferation inhibition rate in the si-LINC00355+ anti-miR-494-3 p group was significantly reduced(P<0.05),and the number of migrating cells and invading cells were significantly increased(P<0.05),the levels of P21,E-c

关 键 词：LncRNA LINC00355 miR-494-3p CCND2 前列腺癌 增殖 侵袭 迁移 

分 类 号：R737.25[医药卫生—肿瘤]