p53基因转染对卵巢癌MDR细胞药物敏感性及恶性表型的影响Influence of drug sensitivity and malignant phenotype of ovarian cancer MDR cells transfected with p53 gene
刘春生,孔北华,马道新,姜洁,李学汤
摘要(Abstract):
研究 p53基因导入已知内源背景的肿瘤 MDR细胞所致的恶性表型和MDR表型的改变及两者的关系。方法:用磷酸钙沉淀法将含有野生型及全长反义 p53cDNA的逆转录病毒载体pDWp53及 pDAp53转染病毒包装细胞 PA317,测定病毒滴度。用此病毒感染卵巢癌多药耐药细胞株A2780/ADM,Southern Blot鉴定,检测转导基因后细胞株的恶性度、多药耐药性等情况。结果:野生型及反义全长p53cDNA均转入 PA317细胞获得效价为(1- 1.5) X105CFU/ml的前病毒,以此感染卵巢癌多药耐药细胞株A2780/ADM,Southem Blot证实p53基因导入该细胞并整合到基因组DNA中,进一步测试观察到:①导入野生型p53基因的A2780/ADM细胞生长被抑制、恶性度降低,细胞形态和生长曲线改变,软琼脂集落形成率及裸鼠接种成瘤率降低;②细胞多药耐药性减弱,对ADM耐药性下降,P-gp表达降低;③反义p53的导入也对A2780/ADM恶性度有一定的影响;④野生型p53导致的MDR细胞恶性表型与MDR水平的降低似有平行关系。结论:p53基因对肿瘤细胞mdr- 1基因的表达可能起调控作用,p53发生突变的 M?
关键词(KeyWords): 基因;p53;多药耐药细胞;药物敏感性测定;恶性表型
基金项目(Foundation):
作者(Author): 刘春生,孔北华,马道新,姜洁,李学汤
DOI: 10.13283/j.cnki.xdfckjz.2000.06.014
参考文献(References):
- 1J.萨姆布鲁克,E.F.弗里奇,T.曼尼阿蒂斯,等.分子克 隆实验指南.第2版.金冬雁等,译,北京:科学出版社, 1992,17—21
- 2 Parker BA, Stark GR.Regulation of simian virus 40 transcrip-tion: Sensitive analysis of the RNA species present early in in- fections by virus or viral DNA. J Viral, 1979, 31(2):360
- 3 Rosenberg J, Amrani DL. A rapid method of southern blot analysis using polyacraylamide gel electrophoresis and vacuumblotting transfer techniques Biotechniques, 1989, 7(1):24
- 4 Feinberg AP,Vogelstein B. A technique for radiolabeling DNArestriction endonuclease fragments to high specifec activity.Anal Biochem, 1983, 132(1):6
- 5 Goldstein LJ, Galski H, Fojo A, et al. Expression of a multi- drug resistance gene in human cancers. J Natl Cancer Inst,1989, 81(2): 116
- 6 Raycroft L,Wu H, Lozano G. Transcriptional activation bywile-type but not transforming mytants of the p53 anti -on- cogen Science, 1990,249(4972):1049
- 7 Miller AD. Use of retriviral vectors for geng transfer and ex- pression . Blood, 1990,76:271
- 8 Marchenko ND, Zaika A, Moll UM.Death signal - inducedlocalization of p53 protein to mitochondria. A potential role in apoptotic signaling. J Biol Chem, 2000, 275(21): 16202