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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?
To study the p53 gene into the known endogenous tumor MDR cells caused by malignant phenotype and MDR phenotype changes and the relationship between the two. Methods: The retroviral vectors pDWp53 and pDAp53 containing wild-type and full-length antisense p53 cDNA were transfected into PA317 cells by calcium phosphate precipitation method to determine the virus titer. The virus was used to infect multidrug resistant ovarian cancer cell line A2780 / ADM and identified by Southern Blot. The malignant degree and multidrug resistance of the cell line were detected after transfection. Results: The wild-type and antisense full-length p53 cDNAs were transfected into PA317 cells to obtain the pro-virus titer of (1- 1.5) X105CFU / ml, which infected the multidrug resistant ovarian cancer cell line A2780 / ADM, Southem Blot It was confirmed that the p53 gene was introduced into the cell and integrated into the genomic DNA. Further tests showed that: ① the growth of A2780 / ADM cells transfected with the wild-type p53 gene was inhibited, the malignancy was decreased, the cell morphology and growth curve were changed, the rate of soft agar colonization and The tumorigenicity of nude mice inoculated was reduced; ②the multidrug resistance of cells was weakened, the resistance to ADM was decreased and the expression of P-gp was decreased; ③the introduction of antisense p53 also had a certain influence on the malignancy of A2780 / ADM; P53-induced MDR malignant phenotype and MDR levels seem to have a parallel relationship. Conclusion: The p53 gene may play a regulatory role in the expression of mdr-1 gene in tumor cells. M?