miR-106a-5p调控 PTEN对鼻咽癌细胞SUNE2增殖和迁移的抑制作用研究

目的研究miR-106a-5p对鼻咽癌细胞SUNE2增殖和迁移的影响。 方法将体外培养的鼻咽癌细胞SUNE2分成对照组（细胞未做任何处理）、Anti-NC组（转染阴性对照抑制剂）、Anti-miR-106a-5p组（转染miR-106a-5p抑制剂）、后期实验另设Anti-miR-106a-5p-inhibitor+si-NC组（转染miR-106a-5p抑制剂、siRNA阴性对照）、Anti-miR-106a-5p-inhibitor+si-PTEN组（转染miR-106a-5p抑制剂、PTEN siRNA），采用Realtime PCR测定miR-106a-5p表达，MTT法检测增殖，Transwell小室检测细胞侵袭和迁移能力变化，用Western blot方法测定vimentin、E-cadherin蛋白表达。在线靶基因预测软件预测miR-106a-5p的靶基因可能为PTEN，双荧光素酶报告系统鉴定miR-106a-5p和PTEN的靶向关系。两组间比较用独立样本t检验，多组间比较采用单因素方差分析，组间两两比较采用LSD-t检验。 结果与正常鼻咽上皮细胞NP69比较，鼻咽癌细胞CNE-2、HK1、SUNE2中miR-106a-5p水平（1.00±0.11比1.84±0.13、2.19±0.14、2.87±0.25）升高，差异具有统计学意义（P < 0.05）。与对照组、Anti-NC组比较，Anti-miR-106a-5p组鼻咽癌细胞miR-106a-5p水平（1.00±0.10、0.99±0.12比0.76±0.08）降低，OD值（0.56±0.05、0.57±0.04比0.32±0.02），细胞侵袭数（128.47±11.65）个、（129.84±10.93）个比（85.12±6.75）个]，迁移数（182.51±14.81）个、（180.68±17.64）个比（122.01±11.62）个]，vimentin蛋白表达水平（0.84±0.09、0.82±0.07比0.30±0.05）降低，E-cadherin蛋白表达水平（0.29±0.04、0.28±0.05比0.76±0.08）升高，差异具有统计学意义（P均< 0.05）。与Anti-miR-106a-inhibitor+si-NC组比较，Anti-miR-106a-inhibitor+si-PTEN组细胞OD值（0.33±0.03比0.52±0.05）、侵袭数（84.16±5.91）个比（105.79±8.63）个]、迁移数（118.42±10.25）个比（164.28±12.05）个]、vimentin蛋白表达水平（0.34±0.06比0.68±0.05）均升高，E-cadherin蛋白表达水平（0.72±0.06比0.29±0.05）降低，差异具有统计学意义（P均< 0.05）。 结论miR-106a-5p可通过靶向调控PTEN抑制鼻咽癌细胞SUNE2增殖和迁移潜能。

Inhibitory of miR-106a-5p regulating PTEN on the proliferation and migration of nasopharyngeal carcinoma cells SUNE2

ObjectiveTo investigate the effect of miR-106a-5p on the proliferation and migration of nasopharyngeal carcinoma cells SUNE2. MethodsThe nasopharyngeal carcinoma cells SUNE2 cultured in vitro were divided into Control group (without treatment), Anti-NC group (transfected with inhibitor control) and Anti-miR-106a-5p group (transfected with miR-106a-5p inhibitor), as well as Anti-miR-106a-5p+si-NC group (transfected with miR-106a-5p inhibitor and siRNA control) and Anti-miR-106a-5p+si-PTEN group (transfected with miR-106a-5p inhibitor and PTEN siRNA). Realtime PCR was used to determine the expression of miR-106a-5p, MTT and Transwell were used to detect proliferation rate, invasion and migration ability of cells, respectively. The expression levels of vimentin and E-cadherin protein were detected by Western blot. The online target gene prediction software suggests that the target gene of miR-106a-5p may be PTEN, and the dual luciferase reporter system examines the targeting relationship between miR-106a-5p and PTEN. The independent sample t-test was used for the comparison between two groups, the ANOVA analysis was used for the multiple group comparison, and the LSD-t test was used for the pairwise comparison of the components. ResultsCompared with NP69 in normal nasopharyngeal epithelial cells, the expression level of miR-106a-5p in nasopharyngeal carcinoma cells CNE-2, HK1, and SUNE2 were increased (1.00±0.11 vs 1.84±0.13, 2.19±0.14, 2.87±0.25), the difference was statistically significant (P < 0.05) .Compared with the Control and Anti-NC group, the expression level of miR-106a-5p (1.00±0.10, 0.99±0.12 vs 0.76±0.08), the OD values (0.56±0.05, 0.57±0.04 vs 0.32±0.02), cell invasion (128.47±11.65, 129.84±10.93 vs 85.12±6.75), the number of migrations (182.51±14.81, 180.68±17.64 vs 122.01±11.62) as well as the expression level of vimentin protein (0.84±0.09, 0.82±0.07 vs 0.30±0.05) of nasopharyngeal cancer cells in the Anti-miR-106a-5p group were significantly reduced. While the expression level of E-cadherin protein was increased (0.29±0.04, 0.28±0.05 vs 0.76±0.08) (P < 0.05). Compared with the Anti-miR-106a-inhibitor+si-NC group, in the Anti-miR-106a-inhibitor+si-PTEN group, the OD value (0.33±0.03 vs 0.52±0.05), the number of invasions (84.16±5.91 vs 105.79±8.63) and migrations (118.42±10.25 vs 164.28±12.05) as well as the expression of vimentin protein (0.34±0.06 vs 0.68±0.05) were all significantly increased. While the expression of E-cadherin protein was decreased (0.72±0.06 vs 0.29±0.05) (P < 0.05) . ConclusionmiR-106a-5p could inhibit the proliferation and migration of nasopharyngeal carcinoma cells SUNE2 by targeted regulation of PTEN.