沉默INHBA-AS1靶向miR-335-3p抑制外阴鳞状细胞癌细胞增殖、迁移

目的:探讨沉默长链非编码RNA（long non-coding RNA，lncRNA）INHBA反义RNA1（INHBA antisense RNA1，INHBA-AS1）对外阴鳞状细胞癌细胞SW954生物学行为的影响及机制。方法:37例外阴鳞状细胞癌组织中INHBA-AS1、miR-335-3p表达运用qRT-PCR检测，高迁移率族蛋白2（high mobility group AT-hook 2，HMGA2）蛋白的表达运用Western blotting检测。实验分为con组、si-NC组、si-INHBA-AS1组、miR-NC组、miR-335-3p mimic组、si-INHBA-AS1+miR-335-3p inhibitor组。应用CCK-8实验检测SW954细胞增殖抑制率，克隆形成实验检测SW954细胞克隆形成，Transwell实验和划痕实验检测SW954细胞迁移，流式细胞术检测SW954细胞凋亡。双荧光素酶报告实验鉴定INHBA-AS1与miR-335-3p、miR-335-3p与HMGA2的靶向关系。结果:外阴鳞状细胞癌组织中INHBA-AS1、HMGA2蛋白的表达水平远高于癌旁组织，miR-335-3p的表达水平低于癌旁组织（P＜0.05）。转染si-INHBA-AS1沉默INHBA-AS1或转染miR-335-3p mimic过表达miR-335-3p后，miR-335-3p表达水平、SW954细胞抑制率及凋亡率升高，HMGA2蛋白的表达水平、SW954细胞克隆形成数、迁移细胞数及划痕愈合率降低（P＜0.05）。INHBA-AS1靶向miR-335-3p，miR-335-3p靶向HMGA2。沉默INHBA-AS1处理的SW954细胞增殖、迁移、凋亡的影响被抑制miR-335-3p所逆转。结论:沉默外阴鳞状细胞癌细胞SW954中的INHBA-AS1通过miR-335-3p靶向HMGA2，抑制SW954细胞增殖、迁移，并诱导其凋亡。

Silenced INHBA-AS1 inhibits proliferation,migration of vulvar squamous cell carcinoma cells by targeting miR-335-3p

Objective:To investigate the effect and mechanism of silencing long non-coding RNA (lncRNA) INHBA antisense RNA1 (INHBA-AS1) on the biological behavior of vulvar squamous cell carcinoma SW954 cells.Methods:In 37 cases of vulvar squamous cell carcinoma tissues,the expression of INHBA-AS1 and miR-335-3p was detected by qRT-PCR,and the expression of high mobility group AT-hook 2 (HMGA2) protein was measured by Western blotting.The experiment was divided into con group,si-NC group,si-INHBA-AS1 group,miR-NC group,miR-335-3p mimic group,and si-INHBA-AS1+miR-335-3p inhibitor group.The CCK-8 experiment was used to assay the proliferation inhibition rate of SW954 cells.The colony formation assay was applied to determine the colony formation of SW954 cells.The Transwell experiment and the scratch experiment were employed to monitor the migration of SW954 cells,and the apoptosis of SW954 cells was analyzed by flow cytometry.The dual luciferase reporter experiment identified the targeting relationship between INHBA-AS1 and miR-335-3p,miR-335-3p and HMGA2.Results:The expression level of INHBA-AS1 and HMGA2 protein in vulvar squamous cell carcinoma tissues was much higher than that of adjacent tissues,and the expression level of miR-335-3p was lower than that of adjacent tissues (P＜0.05).After transfecting si-INHBA-AS1 to silence INHBA-AS1 or transfecting miR-335-3p mimic to overexpress miR-335-3p,the expression level of miR-335-3p,the inhibition rate and apoptosis rate of SW954 cells increased,and the expression level of HMGA2 protein,the number of SW954 cells clones,the number of migrating cells and the healing rate of scratches reduced (P＜0.05).INHBA-AS1 targeted miR-335-3p,and miR-335-3p targeted HMGA2.The effect of silencing INHBA-AS1 on the proliferation,migration and apoptosis of SW954 cells was reversed by the inhibition of miR-335-3p.Conclusion:Silencing INHBA-AS1 in vulvar squamous cell carcinoma SW954 cells through miR-335-3p targeting HMGA2,inhibits the proliferation and migration of SW954 cells,and induces its apoptosis.