目的:探讨采用RNA干扰技术沉默端粒重复序列结合因子2(TRF2)基因对喉鳞状细胞癌Hep-2细胞生长增殖的作用情况。方法:构建特异性表达TRF2mRNA且含有荧光素基因的shRNA真核表达质粒pshRNA,并将其转染Hep-2细胞,采取PCR琼脂糖凝胶电泳检验靶基因TRF2表达情况,流式细胞仪检测细胞凋亡,cell counting kit-8(CCK-8)法观察细胞增殖活性。结果:Hep-2细胞被转染后,TRF2基因表达明显受抑,细胞凋亡发生,增殖活性明显降低。结论:RNA沉默TRF2可明显抑制人喉鳞状癌细胞的增殖,促进细胞发生凋亡。
Objective:To investigate the effect of RNA interference silencing telomere repeat factor 2 by observing Hep-2 cells' proliferation and apoptosis in larynx carcinoma cell line.Method:A recombinant plasmid containing a single shRNA(shTRF2) was constructed.The expression of TRF2 gene was detected by PCR and cell proliferation was examined using CCK-8.Hep-2 cells' apoptosis was detected by flow cytometer (FCM).Result:After treatment with shTRF2,the expression of TRF2 was distinctively depressed,and Hep-2 cells proliferation was obviously inhibited.Compared with control and negative group,cells with treatment of RNAi exhibited significantly more apoptosis.Conclusion:Using RNA interference technique to silence TRF2 gene is effective on inhibiting cancer cells' proliferation and help to induce cancer cells' apoptosis.