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

ABSTRACT

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 .