前列腺癌细胞PC-3M对外源基因pPSA-P21表达的抑制作用

Inhibition effects of the PC-3M cells on expression of exogenous p21 gene

目的:观察外源p21WAF-1/CIP1p21基因在前列腺癌细胞PC-3M中的表达及对细胞增殖和凋亡的影响。方法:构建前列腺组织特异性表达载体pPSA-p21,用lipofectin转染PC-3M细胞,MTT法检测转染前后的细胞生长情况,转染后72h收集细胞进行Westernblotting检测P21蛋白的瞬时表达,并用流式细胞技术(FCM)分析细胞周期和检测细胞P53蛋白的表达。结果:转染后的细胞生长并未受到明显抑制,转染后72hWesternblotting未检测到P21蛋白,FCM分析显示PC-3M细胞多处于G0～G1期和S期,野生型P53和突变型P53蛋白均未见明显表达。结论:PC-3M细胞对外源p21的表达有抑制作用。

Objective: To observe the effects on the proliferation and apoptosis of pc-3M cells after transfected by pPSA-P21 expression vector. Methods: The prostate tissue-specific expression vector pPSA-p21 was constructed and transfected into PC-3M cells with lipofectin. The cell growth was assayed by MTT assay before and after transfection. 72 hours after transfection, the cells were collected and western blotting perfermed to detect the transient expression of P21 protein. And flow cytometric (FCM) analysis was used to analyze cell cycle and detect the expression of P53 protein. Results: The cell growth after transfection was not inhibited obviously. 72 hours after transfection, there was no P21 protein expression to be examined by Western blotting. FCM analysis indicated that most cells were in G0-G1 or S phrase, and neither wild nor mutant P53 protein was found in PC-3M cells. Conclusion: The PC-3M cells can inhibit the expression of exogenous p21 gene.