摘要
目的 探讨反义转化生长因子 β1(TGFβ1)对人膀胱癌细胞体内外增殖的抑制作用。方法 将携TGFβ1反义基因的逆转录病毒载体pRevTβ AS转染膀胱癌EJ细胞 ,观察转染细胞体外及SCID小鼠皮下生长情况 ,通过免疫组化方法评价细胞增殖活性的改变 ;通过病理图像分析和电子显微镜技术观察体内生长的EJ细胞形态学和超微结构改变 ;采用流式细胞分析技术观察转染后细胞周期时相分布的变化。结果 pRevTβ AS能有效抑制EJ细胞TGFβ1mRNA转录和蛋白表达。体外实验表明 ,转染反义TGFβ1可使EJ细胞体外增殖活性和S期细胞比例明显降低 ;体内实验证实 ,反义组种植肿瘤的形成和生长被有效抑制 ,DNA异倍体细胞比例和基因复制活跃程度明显低于对照组。结论 转染反义TGFβ1能明显减弱EJ细胞致瘤性 。
Objective To investigate the inhibitory effects of antisense TGF β1 on proliferation of human bladder transitional cell carcinoma in vitro and in vivo. Methods Human bladder carcinoma cell line EJ was transfected with pRevT β- AS, a replication defective retroviral vector carried antisense TGF β1 fragment . The growth of the transfected cells was observed in vitro and in vivo. TGF β1 mRNA expression and protein expression were detected by RT-PCR and ELISA. The prolif e rative activity was evaluated by immunohistochemistry method. The ultrastructure of cells was observed by image analysis system and electron microscopy. Cell cy cle was determined by flow cytometry. Results The expression of TGF β1 mRNA and protein in EJ cells was inhibited by pRevT β-AS, G 1 to S transition was rest rai ned in cell cycle and cell proliferation decreased in vitro. The tumorigenesis a nd growth of EJ cells and DNA heteroploidy were reduced by antisense TGF β1 in vivo. Conclusion TGF β1 plays a role in vitro proliferation and in vivo growth of bl adder transitional cell carcinoma.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2004年第1期18-21,共4页
Chinese Journal of Oncology
基金
天津市高等学校科技发展基金资助项目 ( 2 0 0 30 31 1 )
