抑癌基因p27对肾癌细胞系GRC-1生长的抑制作用

目的：探讨p27基因对肾癌GRC－1细胞系生长的抑制作用，为肾癌发生机制的研究和基因治疗提供理论依据。方法 采用基因转染技术，将p27 cDNA转染到肾癌GRC－1细胞系中，了解其对细胞增殖能力及细胞周期的影响。结果 转染p27基因的肾癌细胞生长受到抑制，其在软琼脂上克隆形成能力降低了大约4倍。对细胞周期分析表明，处在G0－G1期的细胞由32．2％增加到66．9％，经Dot blot和Western blot杂交证实，p27 mRNA表达有明显差异（P＜0．01），p27的蛋白表达量有明显差异（P＜0．01），说明p27蛋白具有生长抑制功能，其作用点为G1－S。结论 提高肾癌细胞中p27的表达，能抑制肿瘤细胞的生长，导入p27基因是一种治疗肾癌的新途径。

p27 gene suppresses growth of human renal carcinoma GRC-1 cells

AIM To explore the effect of p27 gene on the growth inhibition of renal carcinoma cell line GRC 1.METHODS The p27 cDNA was transfected into human renal carcinoma cell line GRC 1 cells with lipofectamine. The growth rate, cell cycles and colony formation rate of transfected cells were observed by means of MTT and FCM assay. RESULTS Expression of p27 in GRC 1 was identified by Dot blot and Western blot analyses. The growth rate of GRC 1 transfected with p27 gene was markedly suppressed. Colony formation in soft agar was also decreased significantly. Cell cycle analysis by flow cytometry show that the number of cells in G 0 G 1 phase of GRC 1 cells was significantly increased while cells in S and G 2+M phase was decreased compared to that of the control GRC 1 cells. CONCLUSION Transduction of p27 gene into lower expression cancer cells can restore its suppressive effect on cell growth by arrest of cell cycle at G 1 phase.