摘要
目的 :探讨野生型p53基因转染对人卵巢癌细胞株SK0V 3的体外生长及裸鼠体内致瘤性抑制作用 ,并了解 p53基因与SKOV 3细胞对顺铂敏感性之间的关系。方法 :利用脂质体介导 ,将含有人野生型p53cDNA的真核表达质粒pCB6 p53导入SKOV 3细胞中 ,观察该细胞体外生长和裸鼠体内致瘤性改变 ;应用流式细胞仪检测细胞周期及凋亡情况 ,MTT法检测细胞药物敏感性改变。结果 :免疫组化法证实外源性 p53基因在阳性细胞克隆稳定存在 ;转染野生型p53基因的SKOV 3细胞体外生长速率下降 ,裸鼠体内致瘤性丧失 ;G1/G0 期细胞百分比(81.5%)和凋亡细胞百分比(11 %)增高 ;p53表达阳性的SKOV 3细胞对顺铂的敏感性明显增强。结论 :野生型p53基因转染能使SKOV 3细胞出现生长抑制和对顺铂的化疗敏感性增强。
Objective:To investigate the effects of human wild-type p53 gene transfection on the growth and Chemosensitivity of human ovarian cell line SKOV-3 in-vitro.Methods:Recombinant eukaryotic expression vector PCB6-p53 containing full-length human wild-type p53 cDNA was introduced into SKOV-3 cell line by lipofectamine reagent.The expression of p53 was detected by immunohistochemisty;the cell growth and the tumorigenicity of nude mice were observed;the changes of cell cycle and the apoptosis were studied by flow cytometer;the cell proliferation was measured by methyl thiazolyl tetrazolium treated with cisplatin.Results:Exogenous wild-type p53 gene was transfected into SKOV-3 cell line successfully.The stable overexpression of p53 protein in transfected tumors cells was detecteel by immunohistochemisty.The cell growth rate of SKOV-3 cell line transfceted wild-type p53 gene was decrease;the tumorigenicity of nude mice of SKOV-3 cell line transfected wild-type p53 gene was lost.The percentage of phase G1-G10(81.5%)and that of apoptosis (11.0%)of SKOV-3 cells transfected with P CB6-p53 in- creased.The positive expresion of wild-type p53 in SKOV-3 cell line was more sensitive to cisplatin than control cell lines.Conclusion:Our results indicate that the exogenous wild-type p53 expression not only induced suppression of cell growth,but also increased chemosensitivity of human ovarian cancer cells in-vitro.
出处
《四川肿瘤防治》
2001年第3期134-137,共4页
Sichuan Journal of Cancer Control
基金
国家自然科学基金资助项目(基金编号:39470721)
关键词
P53基因
转染
顺铂
卵巢癌
Ovarian carcinoma,p53 gene,Transfection,Cisplatin.
