目的:探讨脂质体介导的TGF-β1基因(pEGFP-TGF-β1)转染诱导人晶状体上皮细胞系-B3(HLEC-B3)细胞周期蛋白激酶抑制因子p21及细胞周期蛋白激酶CDK2的表达及其对细胞周期调控的影响。方法:将pEGFP-TGF-β1转染人晶状体上皮细胞系-B3(HLEC-B3),采用RT-PCR和Western-blot方法检测pEGFP-TGF-β1转染后24,48,72,96hp21,CDK2和Smad4mRNA和蛋白的表达,设立正常细胞组及pEGFP-C2转染组为对照组。结果:pEGFP-TGF-β1转染组TGF-β124h开始升高,48h达到最高,72h略有减少,96h明显减少,但仍高于正常组,而空载体组与正常细胞组则无明显变化。p21变化趋势和TGF-β1相符,CDK2组变化趋势则正好相反,Smad4组48h明显升高,72h下降明显,96h基本恢复正常。结论:TGF-β1通过活化凋亡基因p21,降低细胞周期蛋白激酶CDK2。

·AIM:To investigate the change of p21,CDK2 and Smad4 on the HLEC transfected with pEGFP-TGF-β1 plasmid and the effect of TGF-β1gene transfection on the cell cycle control of human lens epithelial cell.·METHODS:Immortalized human lens epithelial cellline(HLEC-B3)was transfered by pEGFP-TGF-β1 gene.The expression of TGF-β1mRNA,p21mRNA,CDK2mRNA and Smad4 mRNA and protein induced by transfered by TGF-β1 gene were measured by RT-PCR and Western-blot at different time periods.·RESULTS:RT-PCR detected that TGF-β1mRNA began to raise after 24h,culminated after 48h,decreased after 72h,decreased much after 96h,but still more than the normal.p21mRNA companied with it.but CDK2 decreased with it.Western-blot detected that the consequence consisted with RT-PCR.The effect of TGF-β1 gene was demonstrated from the lever of transcription and translation.Smad4 protein raised with the lever of TGF-β1 gene at various time point,but disappeared very quickly.·CONCLUSION:TGF-β1 can increase p21,but decrease CDK2,to impact the cell cycle of HLEC.TGF-β/Smad4 pathway take part in the course.·

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