miR-503靶向作用RECK调控口腔鳞状细胞癌细胞增殖、侵袭和凋亡的机制研究  

miR-503 targeting RECK regulates the proliferation,invasion and apoptosis of oral squamous cells carcinoma cells

作　　者：周新谊 叶涛 邱凤美 ZHOU Xinyi;YE Tao;QIU Fengmei(Department of Stomatology,Jingmen Second People's Hospital,Jingmen 448000,China)

机构地区：[1]荆门市第二人民医院口腔科,荆门448000

出　　处：《口腔颌面外科杂志》2023年第5期305-313,共9页Journal of Oral and Maxillofacial Surgery

摘　　要：目的:通过研究miR-503靶向回复引导半胱氨酸丰富蛋白Kazal基元(reversion inducing cysteine rich protein with Kazal motifs,RECK)在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)进展中的作用。方法:将人OSCC细胞SCC-4分为NC inhibitor组(转染miR-503 inhibitor阴性对照序列)、miR-503 inhibitor组(转染miR-503 inhibitor)、si-NC组(转染RECK siRNA阴性对照序列)、si-RECK组(转染RECK siRNA)、miR-503 inhibitor+si-NC组(共转染miR-503 inhibitor和RECK siRNA阴性对照序列)和miR-503 inhibitor+si-RECK组(共转染miR-503 inhibitor和RECK siRNA)。实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测miR-503和RECK在各组SCC-4细胞及永生化人口腔角质形成细胞RT7中的表达;Western blotting检测RECK蛋白的表达;TargetScan预测miR-503的靶基因并通过双荧光素酶报告基因检测来验证miR-503与RECK的靶向关系;CCK-8和EdU染色检测各组细胞的增殖能力;Transwell小室检测细胞侵袭能力;流式细胞术检测各转染组细胞凋亡情况。结果:相比RT7细胞,miR-503在SCC-4细胞中高表达(P<0.05),RECK则呈低表达(P<0.05);与NC inhibitor组相比,miR-503 inhibitor组细胞中RECK mRNA及RECK蛋白的表达水平均显著升高(P<0.05);TargetScan数据库及荧光素酶报告基因分析显示了RECK和miR-503之间的靶向关系(P<0.01),提示miR-503可靶向抑制RECK的表达;与NC inhibitor组相比,miR-503 inhibitor组SCC-4细胞的增殖能力、侵袭能力均显著下降(P<0.05),而细胞的凋亡则显著增加(P<0.01),抑制miR-503表达降低了OSCC细胞的增殖、侵袭并加速凋亡;与si-NC组相比较,si-RECK组细胞增殖、侵袭能力均显著增加(P<0.05),而细胞凋亡能力显著下降(P<0.05),敲低RECK增加了OSCC细胞的增殖、侵袭并降低凋亡;与miR-503 inhibitor+si-NC组相比较,miR-503 inhibitor+si-RECK组细胞中RECK mRNA的表达水平和细胞凋亡能力均显著下降(P<0.05),细胞增殖、侵袭能力显著增加(P<0.05Objective:To study the role of miR-503 targeting reversion inducing cysteine rich protein with Kazal motifs(RECK)in the progression of oral squamous cell carcinoma(OSCC).Methods:Human OSCC cells SCC-4 were divided into NC inhibitor group(transfected with miR-503 inhibitor negative control sequence),miR-503 inhibitor group(transfected with miR-503 inhibitor),si-NC group(transfected with RECK siRNA negative control sequence),si-RECK group(transfected with RECK siRNA)and miR-503 inhibitor+si-NC group(co-transfected with miR-503 inhibitor and RECK siRNA negative control sequence)and miR-503 inhibitor+si-RECK group(co-transfected with miR-503 inhibitor and RECK siRNA);realtime quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of miR-503 and RECK in immortalized human oral keratinocytes RT7 and SCC-4 cells in each group;Western blotting was used to detect the protein expression level of RECK.TargetScan was used to predict the target gene of miR-503 and dual luciferase reporter gene detection was used to verify the targeting relationship between miR-503 and RECK;CCK-8 and EdU staining were used to detect cell proliferation ability in each group;Transwell chamber was used to detect cell invasion ability;flow cytometry was used to detect cell apoptosis in each transfected group.Results:Compared with RT7 cells,the expression level of miR-503 in SCC-4 cells was increased(P<0.05),while the expression level of RECK was decreased(P<0.05);Compared with NC inhibitor group,the mRNA expression level of RECK and the protein expression level of RECK in the miR-503 inhibitor group were significantly increased(P<0.05).In addition,TargetScan database and luciferase reporter gene analysis showed a targeting relationship between RECK and miR-503(P<0.01),suggesting that miR-503 could target the expression of RECK;compared with NC inhibitor group,the proliferation ability and invasion ability of SCC-4 cells in miR-503 inhibitor group were significantly decreased(P<0.05),while cell apoptosis ability was

关 键 词：miR-503 回复引导半胱氨酸丰富蛋白Kazal基元 靶向调控 口腔鳞状细胞癌 增殖 侵袭 凋亡 

分 类 号：R782[医药卫生—口腔医学]