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为研究野生型P53基因导入诱导血管平滑肌细胞P21基因的表达,探讨P53基因调节细胞周期进程的作用机理,体外培养了人脐动脉平滑肌细胞。将野生型P53基因导入细胞后,应用逆转录-聚合酶链反应半定量测定P21mRNA水平,以免疫组织化学法观察P21蛋白表达的变化,并用流式细胞术分析细胞周期。结果发现,正常生长的血管平滑肌细胞中P21mRNA水平较低,用免疫组织化学法检测不到P21蛋白。野生型P53基因导入并在平滑肌细胞中表达后,显著增加了P21mRNA水平,在免疫组织化学检测中呈现很强的阳性显色反应、引起平滑肌细胞停滞在G0/G1期。以上结果提示,野生型P53基因通过诱导P21基因表达调控血管平滑肌细胞周期。
In order to study the expression of P21 gene induced by wild-type P53 gene in vascular smooth muscle cells and the mechanism of P53 gene regulating cell cycle progression, human umbilical artery smooth muscle cells were cultured in vitro. After the wild-type p53 gene was introduced into the cells, the level of P21 mRNA was semi-quantitatively determined by reverse transcription-polymerase chain reaction. The expression of P21 protein was observed by immunohistochemistry. Cell cycle was analyzed by flow cytometry. The results showed that P21 mRNA in normal vascular smooth muscle cells was low, and P21 protein was undetectable by immunohistochemistry. After the wild-type p53 gene was introduced and expressed in smooth muscle cells, the level of P21 mRNA was significantly increased, which showed a strong positive color reaction in immunohistochemical examination, resulting in the arrest of smooth muscle cells in G0 / G1 phase. The above results suggest that wild-type P53 gene regulates vascular smooth muscle cell cycle by inducing P21 gene expression.