慢病毒介导的miRNA-197-3p-siRNA对喉癌细胞系Hep-2生物学行为的影响

The effect of miRNA-197-3p-siRNA-1 entivirus on biological behaviors in human laryngeal carcinoma cell line Hep-2

目的通过构建慢病毒介导(lentivirus,LV)的miRNA-197-3p-siRNA,探讨下调miRNA-197-3p对喉癌细胞系Hep-2生物学行为的影响。方法针对目标序列合成特异性siRNA干扰片段并克隆入慢病毒载体,将重组miRNA-197-3p-RNAi-LV3转染喉鳞状细胞癌Hep-2。Hep-2细胞设为干扰组(miRNA-197-3p-siRNA,MI组)、阴性对照组(negative control,NC组)和空白对照组(blank,B组);分别采用聚合酶链反应方法检测miRNA-197-3p的表达情况;用CCK8法检测细胞增殖情况,流式细胞仪检测细胞周期及凋亡情况,Transwell侵袭实验检测细胞侵袭情况。结果 miRNA-197-3p-RNAi-LV3可显著降低miRNA-197-3p的表达。CCK8实验显示细胞增殖能力显著降低(P<0.05)。Transwell实验显示细胞侵袭能力明显减弱(P<0.05)。流式细胞仪检测表明转染后对Hep-2细胞周期及细胞凋亡均无明显影响(P>0.05)。结论 miRNA-197-3p-siRNA通过下调miRNA-197-3p的表达,可以抑制Hep-2细胞增殖,并降低其侵袭能力。

Objective To study the effect of lentivirus-mediated miRNA-197-3p-RNAi on biological behaviors in human laryngeal carcinoma cell line Hep-2. Methods miRNA-197-3p specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector.Hep-2 cells were infected by miRNA-197-3p-siRNA （MI） recombinant lentivirus（miRNA-197-3p-siRNA-LV3）.The Hep-2 cells were divided into MI group,negative control （NC） group and blank （B） group in vitro.The expression of the targets of miRNA-197-3p was detected by RT-PCR.Cell proliferation was detected by CCK8 kit.Cell cycle and apoptosis were detected by flow cytometry.The invasion was detected by Transwell test. Results The expression level of miRNA-197-3p was inhibited significantly by miRNA197-3p-siRNA-LV3.The proliferation of Hep-2 was also markedly suppressed in CCK8 （P〈0.05）.The number of invasive cells that migrated through the chamber decreased （P〈0.05）. The cell cycle of Hep-2 and apoptosis had no obvious effect （P〉0.05）. Conclusions miRNA-197-3p specific siRNA can suppress the proliferation and reduce the invasiveness of Hep-2 cells.