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目的:研究hsa-miR-193b对喉鳞状细胞癌Hep-2细胞增殖、凋亡等生物学功能的作用。方法:实验分组为Hep-2组、NC组(Hep-2细胞+miR-193binhibitor NC)、miR-193b抑制组(Hep-2细胞+miR-193binhibitor)。采用qRT-PCR检测miR-193b的表达,MTT法检测细胞增殖,流式细胞仪检测细胞周期以及凋亡,Transwell检测细胞迁移能力。采用生物信息学技术进行靶基因预测,Western blot检测转染miR-193b后Hep-2细胞SMAD3的表达情况。结果:转染miR-193binhibitor后,miR-193b抑制组细胞miR-193b的表达显著降低,与Hep-2组和NC组比较差异有统计学意义(P<0.01),后2组miR-193b表达量差异无统计学意义(P>0.05)。和NC组比较,miR-193b抑制组的Hep-2细胞增殖能力在24、48和72h显著降低(P<0.05或P<0.01),G1/S期的细胞比例增多(P<0.01),细胞凋亡明显增多(P<0.01),细胞迁移能力显著降低(P<0.01)。生物学信息分析预测miR-193b靶基因为SMAD3,Hep-2细胞转染miR-193b后与NC组相比,SMAD3蛋白表达显著下调(P<0.05)。结论:miR-193b抑制剂能够抑制Hep-2细胞的增殖、迁移,促进细胞凋亡,提示miR-193b对喉鳞状细胞癌起到“癌基因”的作用,有望成为临床治疗的新靶点。
OBJECTIVE: To study the effect of hsa-miR-193b on the biological functions of Hep-2 cells such as proliferation and apoptosis in laryngeal squamous cell carcinoma. Methods: The experimental group was divided into Hep-2 group, NC group (Hep-2 + miR-193 binhibitor NC) and miR-193b group (Hep-2 + miR-193 binhibitor). The expression of miR-193b was detected by qRT-PCR. Cell proliferation was detected by MTT assay. Cell cycle and apoptosis were detected by flow cytometry. Transwell assay was used to detect cell migration. The target genes were predicted by bioinformatics technology. The expression of SMAD3 in Hep-2 cells transfected with miR-193b was detected by Western blot. Results: After transfected with miR-193binhibitor, the miR-193b expression in miR-193b group was significantly lower than that in Hep-2 group and NC group (P <0.01), and miR-193b expression in the latter two groups The difference was not statistically significant (P> 0.05). Compared with NC group, the proliferation of Hep-2 cells in miR-193b group was significantly decreased at 24,48 and 72h (P <0.05 or P <0.01), and the proportion of cells in G1 / S phase was increased (P <0.01) Apoptosis was significantly increased (P <0.01), and cell migration was significantly decreased (P <0.01). The biological information analysis predicted miR-193b target gene as SMAD3, and miR-193b transfected Hep-2 cells significantly reduced the expression of SMAD3 protein compared with NC group (P <0.05). Conclusion: The miR-193b inhibitor can inhibit the proliferation, migration and apoptosis of Hep-2 cells, suggesting that miR-193b may play an important role in the oncogenesis of laryngeal squamous cell carcinoma Targets.