circZNF609靶向miR-1287-5p对皮肤鳞状细胞癌A431细胞增殖和凋亡的影响

Effects of circZNF609 Targeting miR-1287-5p on Proliferation and Apoptosis of Cutaneous Squamous Cell Carcinoma A431 Cells

目的探索环状核糖核酸锌指蛋白609(circZNF609)是否通过靶向微小核糖核酸-1287-5p(miR-1287-5p)影响皮肤鳞状细胞癌(cSCC)A431细胞增殖和凋亡。方法应用实时定量聚合酶链反应(qRT-PCR)测定circZNF609和miR-1287-5p在cSCC组织中的表达。在cSCC A431细胞中转染小分子干扰阴性对照(si-NC组)、sicircZNF609(si-circZNF609组)、miR阳性对照(miR-NC组)、miR-1287-5p模拟物(miR-1287-5p组)、si-circZNF609+anti-miR-NC(si-circZNF609+anti-miR-NC组)、si-circZNF609+miR-1287-5p抑制剂(si-circZNF609+anti-miR-1287-5p组)。运用细胞计数试剂盒-8(CCK-8)、克隆形成实验、流式细胞术和蛋白质印迹法(Western blotting)测定细胞活力、克隆形成、凋亡和活化的胱天蛋白酶-3(cleaved caspase-3)、cleaved caspase-9蛋白表达。双荧光素酶报告实验分析circZNF609和miR-1287-5p的靶向关系。结果circZNF609在cSCC组织中的表达水平比癌旁组织增加约2.85倍,miR-1287-5p在cSCC组织中的表达水平是癌旁组织的34%(P<0.05)。干扰circZNF609表达或miR-1287-5p过表达可降低cSCC A431细胞活力、克隆形成数,提高细胞凋亡率和cleaved caspase-3、cleaved caspase-9蛋白表达(P<0.05)。circZNF609靶向调控miR-1287-5p的表达。下调miR-1287-5p表达逆转了干扰circZNF609表达对cSCC A431细胞增殖和凋亡的作用。结论干扰circZNF609表达通过靶向miR-1287-5p,抑制cSCC的细胞增殖,促进其凋亡。

Objective In order to explore whether circular RNA zinc finger protein 609(circZNF609)can affect the proliferation and apoptosis of cutaneous squamous cell carcinoma(cSCC)A431 cells by targeting microRNA-1287-5p(miR-1287-5p).Methods Real-time quantitative polymerase chain reaction(qRT-PCR)was used to determine the expression of circZNF609 and miR-1287-5p in cSCC tissues.Transfect si-NC(si-NC group),si-circZNF609(si-circZNF609 group),miR-NC(miR-NC group),miR-1287-5p mimic(miR-1287-5p group),si-circZNF609 and anti-miR-NC(si-circZNF609+anti-miR-NC group),si-circZNF609 and miR-1287-5p inhibitor(si-circZNF609+anti-miR-1287-5p group)into cSCC A431 cells.Cell counting kit 8(CCK-8),colony formation experiment,flow cytometry and Western blotting were employed to determine cell viability,colony formation,apoptosis and the expression of activation of cleaved caspase-3,cleaved caspase-9 protein.The dual luciferase report experiment analyzed the targeting relationship between circZNF609 and miR-1287-5p.Results The expression level of circZNF609 in cSCC tissues was about 2.85 times higher than that in adjacent tissues.The expression level of miR-1287-5p in cSCC tissues was 34%of that in adjacent tissues(P<0.05).Interference with the expression of circZNF609 or overexpression of miR-1287-5p reduced the viability and the number of colony formation of cSCC A431 cells,and increased the apoptosis rate and the protein expression of cleaved caspase-3 and cleaved caspase-9(P<0.05).circZNF609 targets and regulates the expression of miR-1287-5p.Down-regulating the expression of miR-1287-5p reversed the effect of interfering with the expression of circZNF609 on the proliferation and apoptosis of cSCC A431 cells.Conclusion Interfering the expression of circZNF609 can inhibit the proliferation of cSCC and promote its apoptosis by targeting miR-1287-5p.