RNAi沉默miRNA-224-5p对喉鳞状细胞癌Hep-2细胞增殖、凋亡及侵袭能力的影响

目的通过慢病毒载体介导的RNA干扰技术,来探讨沉默miRNA-224-5p对人喉鳞状细胞癌（简称喉鳞癌）Hep-2细胞增殖、凋亡及侵袭能力的影响。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-224-5p-RNAi-LV3转染喉鳞癌Hep-2。设为干扰组（miRNA-224-5p-siRNA,SI组）、阴性对照组（NC组）和空白对照组（BC组）,分别采用聚合酶链反应方法检测miRNA-224-5p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-224-5p-RNAi-LV3可显著降低miRNA-224-5p的表达,转染后Hep-2细胞增殖能力显著降低,细胞侵袭能力明显减弱,细胞周期及细胞凋亡均未见明显影响。结论 RNAi沉默miRNA-224-5p可以抑制Hep-2细胞增殖,并降低其侵袭能力。

Effect of miRNA-224-5p gene silencing on cell proliferation,apoptosis and invasion in human laryngeal cancer cell line Hep-2 by RNA interference

OBJECTIVE To explore the influence of miRNA-224-5p gene silencing on cell proliferation,apoptosis and invasion in human laryngeal cancer cell line Hep-2 by lentiviral vector-mediated RNA interference.METHODS miRNA-224-5p specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector.Hep-2 cells were infected by miRNA-224-5p-siRNA recombinant lentivirus（miRNA-224-5p-siRNA-LV3）.The Hep-2 cells were divided into SI group,NC group and BC group in vitro.The expression of the targets of miRNA-224-5p 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-224-5p was inhibited significantly by miRNA224-5p-siRNA-LV3.The proliferation of Hep-2 was also markedly suppressed.The number of invasive cells that migrated through the chamber decreased.The cell cycle and apoptosis had no obvious effect.CONCLUSION MiRNA-224-5p gene silencing by RNAi can suppress the proliferation and reduce the invasiveness of Hep-2 cells.