摘要
目的 探讨抑癌基因 p5 3和 Rb对膀胱癌细胞的生长抑制作用。方法 利用逆转录病毒和电穿孔法将 p5 3、Rb单独或联合导入膀胱癌细胞株 T2 4中 ,Northern杂交等证实外源目的基因的表达后 ,观测并比较不同转染后细胞的生长、增殖水平和成瘤性等。结果 获得 p5 3、Rb单独和联合转染的 T2 4细胞 ,Northern杂交等证实 T2 4细胞中 p5 3失活而 Rb正常 ,转入的外源野生型 p5 3基因和 Rb基因均被表达。与未转染的对照组 T2 4细胞相比 ,转入 p5 3的细胞其生长速率明显降低 ,细胞增殖受抑制 ,软琼脂克隆形成能力丧失 ;而只转染 Rb的细胞除软琼脂克隆形成能力有所降低外 ,细胞生长和增殖未受明显抑制。结论 外源野生型 p5 3能抑制 T2 4生长并降低其成瘤性 ,增加 Rb的表达对细胞生长无明显抑制作用。p5 3对 T2 4细胞的生长抑制作用与外源 Rb基因导入与否无明显关系 ,Rb表达水平正常的膀胱癌细胞其异常增殖可能与 Rb蛋白磷酸化状态的关系更为密切。
Objective To investigate the effects of tumor suppressor genes p53 and Rb on the growth, proliferation, and tumorigenicity of bladder cancer cells. Methods We used the retroviral gene transfection and electro perforation method to introduce extrogenous p53 and Rb gene separately or simultaneously into bladder cancer cell line T24. After confirmation of the expression of the extrogenous genes, we analyzed the growth curve, the cell cycle fraction, and the soft agar colony formation of the cancer cells. Results p53 was inactivated but Rb was normal in the T24 cell before transfection. After single transfection or co transfection of p53 and Rb, the expression of extrogenous p53 and Rb was confirmed at the mRNA and protein levels. Compared to control cells without any transfection, the T24 cells transfected with wild type p53 showed obvious decrease in cell growth, suppression of cell proliferation, and loss of capacity of soft agar colony formation. On the contrary, the cells with single transfection of Rb only showed decrease of soft agar colony formation. Conclusion The introduction of extrogenous wild type p53 could suppress the growth, proliferation, and tumorigenicity of T24 cell, but the increased expression of Rb could purely reduce the tumorigenicity of T24 cell.
出处
《华西医科大学学报》
CSCD
北大核心
2002年第4期529-532,共4页
Journal of West China University of Medical Sciences
基金
国家自然科学基金资助 (批准号 3 95 70 70 2 )
