靶向沉默胶质瘤细胞转录因子1基因表达抑制喉癌细胞增殖及其机制

Inhibition effect of targeted silence of Gli1 expression on the growth of Hep-2 human laryngeal squamous carcinoma cells and its mechanism

目的探讨应用RNA干扰技术沉默胶质瘤细胞转录因子1(Glioma-associated Oncogene Homolog 1,Gli1)基因表达对喉癌Hep-2细胞增殖影响极其机制。方法应用脂质体法将Gli1小发夹RNA(small hairpin RNA,shRNA)转染到人喉癌Hep-2细胞,流式细胞术检测转染效率,四甲基偶氮唑蓝(MTT)法检测细胞增殖变化,Western blot法检测Gli1和cyclin D1蛋白表达。结果 Gli1蛋白表达在Gli1 shRNA组与空白对照组(t=15.565,P<0.01)和空质粒对照组(t=14.406,P <0.01)相比明显降低,统计学有显著差异。细胞增殖抑制率在Gli1sh RNA组与空白对照组(t=9.728,P<0.01)和空质粒对照组(t=9.649,P<0.01)相比明显增高,统计学有显著差异。cyclin D1蛋白表达在Gli1 shRNA组与空白对照组(t=23.045,P<0.01)和空质粒组(t=26.630,P<0.01)相比均明显降低。结论 RNA干扰技术沉默Gli1基因表达明显抑制喉癌Hep-2细胞增殖活性,其机制可能是通过降低cyclin D1蛋白表达。

OBJECTIVE To investigate the effect of silencing of Gli1 by RNAi on proliferation and its mechanisms in laryngeal carcinoma cell line Hep-2 cells. METHODS Gli1 shRNA was transfected into Hep-2 cells by lipofectamin assay. The transfection efficiency was detected by f low cytometry. The cell proliferation was detected by MTT assay. The expression of Gli1 and cyclin D1 protein was analyzed by Wester n blot. R ESULTS The expression of Gli1 protein in group Gli1 shRNA was significantly lower than that in blank group and mock group respectively(t1=15.565, t2=14.406,all P <0.01). The growth inhibition rate in group Gli1 shRNA were significantly higher than that in blank group and mock group respectively(t1=9.728, t2=9.649, all P <0.01). The expression of cyclin D1 protein in group Gli1 shRNA was significantly lower than that in blank group and mock group respectively(t 1=23.045, t2=26.630, all P <0.01). CONCLUSION Gli1 shRNA could inhibit the growth of laryngeal carcinoma cell line Hep-2 cells, which mechanism might be inhibited the expression of cyclin D1.