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目的 :建立高表达 β 葡萄糖醛酸苷酶 (以下简称 βG)基因的肾癌细胞模型 ,并观察转染细胞的生物学特性。方法 :构建真核表达载体 pcDNA3.1 βG ,并利用阳离子脂质体介导将其转入人肾癌细胞株GRC 1中。用mRNA打点杂交和Westernblot,鉴定其转录与表达水平 ,并通过光镜、电镜及细胞增殖周期检测等方法 ,观察转染细胞的生物学特性。结果 :在mRNA与蛋白水平证实 ,转染细胞内有 βG基因的高表达。透射电镜观察 ,转染细胞中的溶酶体、内质网丰富 ,细胞微绒毛及突起增多 ,但转染前后肾癌细胞GRC 1的周期及生长情况无显著性差别。结论 :应用脂质体介导法 ,将βG基因导入人肾癌GRC 1细胞后 ,获得生物学特性稳定 βG基因高表达的肾癌细胞模型 ,为进一步研究奠定了基础
OBJECTIVE: To establish a kidney cancer cell model with high expression of β-glucuronidase (hereinafter referred to as βG) gene and to observe the biological characteristics of the transfected cells. Methods: The eukaryotic expression vector pcDNA3.1 βG was constructed and transfected into human renal carcinoma cell line GRC 1 by cationic liposome. The transcription and expression levels were identified by dot blot hybridization and Western blotting. The biological characteristics of transfected cells were observed by light microscopy, electron microscopy and cell cycle detection. Results: The mRNA and protein levels confirmed the high expression of βG gene in transfected cells. Transmission electron microscopy showed that the lysosomes in the transfected cells were rich in endoplasmic reticulum, with increased microvilli and protrusions. However, there was no significant difference in the cell cycle and growth of GRC1 cells before and after transfection. Conclusion: The βG gene was introduced into human renal cell carcinoma cell line GRC-1 by liposome-mediated method, and the model of renal cancer cell with high expression of stable βG gene was obtained, which laid the foundation for further study