野生型p53基因促进长春新碱诱导的细胞凋亡

WILD TYPE p53 STIMULATES VINCRISTINE INDUCED APOPTOSIS

为探讨ＶＣＲ与ｐ５３介导的细胞凋亡的关系，将野生型ｐ５３基因导入一个内源性ｐ５３基因重组的多药耐药细胞系ＫＢＶ２００，经抗性筛选和杂交鉴定获得外源性ｐ５３基因稳定表达的转染细胞系ＫＢＶ２００ｐ５３。ＶＣＲ处理细胞后，经电镜及荧光显微镜观察表明ＫＢＶ２００ｐ５３细胞呈现凋亡的典型形态学特征；而流式细胞术结果显示经６００ｎｍｏｌ·Ｌ－１的ＶＣＲ作用２４ｈ，ＫＢＶ２００ｐ５３细胞和ＫＢＶ２００细胞的凋亡百分数分别为４２４％和８４％。结果提示野生型ｐ５３基因能促进ＶＣＲ诱导的细胞凋亡。

Recent studies indicate that wild type p53 can trigger cell apoptosis induced by many chemotherapeutic agents which induce DNA damage or cause disruptions of DNA metabolism, such as ADM, 5 FU, VP 16 and radiation We introduced the wild type p53 gene into a MDR cell line KB V200 in which the endogenous p53 was found to be rearranged By G418 selection and Northern blot analysis, a G418 resistant clone named KB V200 p53 was obtained which continuously expressed the exogenous wild type p53 mRNA After treatment with Vincritine(VCR), the wild type p53 expression cells presented typical morphology characteristic of apoptosis analysed under electron and fluorescence microscopes Flow cytometer analysis showed that the KB V200 p53 cells were more readily undergo apoptosis than their parental cells KB V200 After treatment with VCR 600 nmol·L -1 for 24 h, the apoptotic percentage of KB V200 p53 and KB V200 cells was about 42 4% and 8 4%, respectively This result indicates that wild type p53 stimulates VCR induced apoptosis in KB V200 cells