沉默 annexin A5对喉癌Hep-2细胞增殖和侵袭的影响

Annexin A5 Silence for Influence on Hep-2 Cell Proliferation and Invasion

目的探讨小干扰RNA(small interference RNA,siRNA)沉默人喉癌Hep-2细胞中的annexin A5基因表达对Hep-2细胞增殖和侵袭的影响研究。方法siDirect Version2.0设计软件设计针对annexin A5基因的特异性siRNA干扰片段及无关序列的siRNA片段各一对,将细胞分成3组,即siRNA干扰组、空白对照组及阴性对照组,用脂质体Lipofectamine^TM 2000将siRNA转染至喉癌Hep-2细胞,转染48h后用实时PCR及Western blot法检测Hep-2细胞干扰后各组annexin A5基因mRNA和蛋白的表达变化;CCK-8实验检测Hep-2细胞增殖情况,Transwell小室侵袭实验检测Hep-2细胞侵袭能力的变化。结果Hep-2 细胞转染后48h,siRNA干扰组annexin A5 mRNA及蛋白的表达较阴性对照组和空白对照组表达均降低(P<0.05),其表达被显著抑制,且有效干扰annexin A5后Hep-2细胞增殖抑制,侵袭能力减弱,阴性对照组和空白组比较,差异有统计学意义(P<0.05)。结论有效沉默annexin A5基因的表达能抑制Hep-2细胞的增殖和侵袭能力,annexin A5基因可能促进喉癌的发生、发展。

Objective To explore the effect of RNA interference(RNAi) targeting annexin A5 influenced the proliferation and invasion of Hep-2 cells.Methods The specially siRNA and negative siRNA fragments of annexin A5 were designed by the siDirect Version2.0,and these fragments were transported by the Lipofectamine TM 2000 to the Hep-2 cells.Hep-2 cells were divided into three groups,which were blank control group,negative control group,and specially siRNA group.RT-PCR and Western blot were used to identify the inhibition efficiency after transfection for 48 hours.Cell counting kit-8 (CCK-8) was used to detect cells proliferation.The Transwell chamber assay was employed to assess cell invasion.Results After cells transfection for 48 hours,the mRNA and protein expression level of annexin A5 were all down-regulated compared to the negative control gronp and the blank control group.After knocking down of annexin A5,the cells proliferation and invasion of the specially siRNA group were inhibited,and the difference was statistically significant which compared to the negative control group and the blank control group.Conclusion After effective silent the expression of annexin A5 in Hep-2 cells,the proliferation and invasion ability of Hep-2 cells were inhibited,annexin A5 may promote the development of laryngeal cancer.