miR-936对人喉癌细胞株Hep-2增殖、迁移、侵袭和药物敏感性的影响

Effects of microRNA-936 on the proliferation,migration,invasion and drug sensitivity of human laryngeal cancer cell line Hep-2

目的探讨微小RNA-936(miR-936)对人类喉癌细胞株Hep-2增殖、迁移、侵袭和药物敏感性的影响。方法通过慢病毒感染构建miR-936过表达的喉癌细胞株Hep-2细胞系(miR-936组)及空载对照组(Vector组)。荧光显微镜观察病毒感染效率,实时荧光定量PCR检测2组细胞miR-936的表达量。MTT增殖实验分析2组细胞增殖能力的变化。MTT药敏实验比较2组细胞药物敏感性的变化。划痕实验和Transwell小室实验分别检测2组细胞迁移和侵袭能力,激光共聚焦扫描显微镜观察细胞骨架变化。结果 miR-936组Hep-2细胞的miR-936 mRNA表达水平高于Vector组(t=5.600,P<0.05)。miR-936组Hep-2细胞增殖能力明显低于Vector组。培养24 h和48 h时miR-936组Hep-2细胞的迁移能力均低于Vector组(P均<0.01);miR-936组细胞的侵袭能力低于Vector组(P<0.01);此外,miR-936过表达后Hep-2细胞的微管微丝与Vector组相比减少。药物敏感实验结果显示miR-936组Hep-2细胞对顺铂和阿霉素的敏感性较Vector组增加(均P<0.05)。结论 miR-936可以抑制Hep-2细胞的增殖、迁移和侵袭,并且能够增加Hep-2细胞对顺铂和阿霉素的敏感性。

Objective To investigate the effect of microRNA-936（miR-936） on the proliferation, migration, invasion and drug sensitivity of human laryngeal cancer cell line Hep-2. Methods The Hep-2 cell line overexpressing miR-936 （miR-936 group） and the empty vector control group （vector group） were constructed by lentivirus infection. The efficiency of virus infection was observed by fluorescence microscope. The expression of miR-936 was detected by RT-qPCR. MTT proliferation assay was used to analyze the changes of cell proliferation in both groups. MTT drug sensitivity assay was performed to compare the changes of drug sensitivity in both groups. Cell migration and invasion ability in two groups was evaluated by Transwell chamber assay. Confocal microscope was adopted to observe the variations in cell cytoskeleton. Results The expression level of miR-936 mRNA of Hep-2 cells in the miR-936 group was significantly higher than that in the vector group （ t=5.600,P 〈0.05 ） . In the miR-936 group, the proliferation capacity of Hep-2 cells was significantly lower compared with that in the vector group. The migration ability of Hep-2 cells in the miR-936 group was significantly lower than that in the vector group at 24 h and 48 h after culture （both P 〈0.01 ） . The cell invasion ability in the miR-936 group was considerably lower than that in the vector group （ P 〈0.01 ） . The quantity of microtubules of Hep-2 cells was significantly reduced after overexpression of miR-936 compared with that in the vector group. Drug sensitivity test demonstrated that the sensitivity of Hep-2 cells to cisplatin and adriamycin was significantly increased in the miR-936 group compared with that in the vector group （ P 〈0.01 ） . Conclusion miR-936 can inhibit the proliferation, migration and invasion of Hep-2 cells, and can increase the sensitivity of Hep-2 cells to cisplatin and adriamycin.