重组人p53腺病毒联合温热顺铂不同用药方式下对人结肠癌细胞株的抑制作用

Sequencial Administration Effect of Warm Cisplatin with Recombinant Adenoviral Human p53 Gene on Human Colon Cell Lines

目的研究rAd-p53与热化疗以不同序贯方式联合应用对不同p53基因状态的结肠癌细胞的抑制作用。方法通过体外细胞培养,采用PT-PCR和Western blot检测验证细胞内不同p53基因状态,利用MTT法检测rAd-p53与42℃热浴顺铂的IC30,以及用药物联合指数计算法观察rAd-p53与42℃热浴顺铂不同用药模式下对不同p53基因状态的结肠癌细胞的抑制作用。结果在p53基因野生型的HCT116+/+细胞中42℃热浴顺铂与rAd-p53同时给药表现为协同作用,42℃热浴顺铂序贯rAd-p53给药模式表现为近似叠加作用,rAd-p53序贯42℃热浴顺铂用药模式表现为拮抗作用。在p53基因敲除的HCT116-/-细胞中,42℃热浴顺铂与rAd-p53同时给药表现为强协同作用,42℃热浴顺铂序贯rAd-p53给药模式表现为协同作用,rAd-p53序贯42℃热浴顺铂用药模式表现为拮抗作用。结论在结肠癌细胞中42℃热浴顺铂与rAd-p53在同时给药的模式下疗效最好,然而不同结肠癌细胞中p53基因状况决定其产生不同作用效果,缺失型p53基因细胞协同效果优于野生型p53基因细胞。

Objective To investigate the best time sequence combined treatment thermochemotherapy and rAd-p53 to the colon cancer cells with p53 gene in different states in vitro. Methods Human colon cancer cells with p53 gene in different states （ HCT116 ＋/＋ and HCT116 -/- ） was used as the research model. RT-PCR and Western blot was used to verify the state of the p53 gene in different human colon cancer cells, and inhibition ratio to colon cancer ceils （HCT116 ＋/＋ and HCT116 -/- ） of thermochemotherapy com- bined with rAd-p53 in different time sequence was assessed by the method of MTT. Results In HCT116 ＋/＋ cells p53 gene in wild-type, the simultaneously administration of thermochemotherapy and rAd-p53 showed a synergistic effect, thermochemotherapy administered sequential rAd-p53 showed similar pattern overlay effect, and tAd-p53 sequential administration of thermochemotherapy model showed antagonism. In HCT116 -/- cells with p53 gene knocked out, the simultaneously administration of thermochemotherapy and rAd-p53 showed a strong synergistic effect, thermochemotherapy administered sequential rAd-p53 showed synergistic effect, and rAd-p53 sequential administration of thermochemotherapy model showed antagonism.Conclusions Thermochemotherapy and rAd-p53 in different sequences administration has different interaction, and its respective of the status of endogenous p53 gene. The simultaneously administration of thermochemotherapy and tAd-p53 have the best synergistic efficacy. Synergistic effect of p53-defieient ceils is better than wildtype p53 gene cells.