摘要
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。
Aim To study the expression of P21 gene in vascu-lar smooth muscle cells(VSMC)induced by introduc-tion of wild-type P53 gene and explore the mechanism of regulation for cell cycle progression by wild-type P53 gene.Methods A P53 gene recombinant adenovirus vec-tor,AdCMVP53.was transfected into the cultured SMC derived from human umbilical artery.The gene expression of P21 was quantified by reverse transcrip-tion-polymerase chain reaction(RT-PCR).The con-tent of P21 was detected by immunochemical staining.Flow cytometry was used for cell cycle analysis.Results The level of P21 mRNA and protein was increased in VSMC transfected with AdCMVP53.Overexpression of wild-type P53 could arrest the SMC at G0/G1 phase of the cell cycle.Conclusion Wild-type P53 gene could induce the expression of P21 gene,leading to repression of cell cycle progression.
出处
《中国动脉硬化杂志》
CAS
CSCD
1998年第4期283-286,共4页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金!39570775
