Skp2基因沉默对 SPC-A-1肺癌细胞增殖和凋亡的影响

目的：探讨 S 期激酶相关蛋白2（Skp2）基因沉默对肺癌细胞增殖和凋亡的影响。方法将 Skp2 RNA 干扰表达载体转染至 SPC-A-1肺癌细胞中，G418筛选获得阳性克隆细胞。通过实时荧光定量（RT-PCR）、蛋白免疫印迹法（Western blot）检测肺癌细胞中 Skp2的表达。采用流式细胞术、四甲基偶氮唑盐（M TT ）法检测各组肺癌细胞生长、凋亡情况。结果转染 Skp2 shRNA 表达载体的肺癌细胞中 Skp2蛋白表达量明显减少，抑制效率分别可达75．3±5．1，70．4±3．2；转染 Skp2 shRNA 表达载体的肺癌细胞生长减慢，阻滞于 G1期的增多，S 期细胞减少。 Skp2 shRNA 转染质粒组凋亡率较阴性对照组明显增加，细胞凋亡率分别为（17．5±2．8）％、（15．6±3．1）％。结论通过特异性沉默 Skp2基因的表达，可有效降低肺癌细胞中 Skp2蛋白表达水平，抑制肺癌细胞生长及增加细胞凋亡。

The effect of Skp2 gene silencing on the proliferation and apoptosis of SPC-A-1 lung cancer cells

Objective To investigate the effect of Skp2 gene silencing on the proliferation and apoptosis of SPC-A-1 lung cancer cells .Methods Specific small hairpin RNA (shRNA ) targeting Skp2 gene was introduced into lung cancer cells by Lipo-fectamine 2000 and the positive clones were screened by G418 .The Skp2 mRNA and protein expression level of lung cancer cells were detected by RT-PCR and Western blot .M TT method and flow cytometry (FCM ) analysis were used to observe the effect of RNAi on proliferation of lung cancer cells .Cell apoptosis was analyzed by FCM .Results Transfected with Skp2 shRNA expression vector significantly reduced the expression of Skp2 protein in lung cancer cells .Inhibition efficiency was respectively (75 .3 ± 5 .1) , (70 .4 ± 3 .2) .P27 protein expression was increased significantly in lung cancer cells .The growth of lung cancer cells transfected with Skp2 shRNA was blocked ,with Gl phase cells increased and S phase cells decreased .The apoptosis rate of cancer cells was higher in Skp2 shRNA groups than in control groups .Apoptosis rates were (17 .5 ± 2 .8)% ,(15 .6 ± 3 .1)% in Skp2 shRNA groups .Conclusion Specific inhibition of the expression of Skp2 can slow down the growth of lung cancer cells ,increase cell apop-tosis .