p53及p21^(waf1)蛋白表达与人舌癌细胞多药耐药性的关系  

Relationship between the expression of p53 and its downstream effecter p21^(waf1) proteins and multi-drug resistance of human tongue cancer cell line

作　　者：何春梅[1] 刘斌[1] 王柯敏[1] 唐红星[1] 何丽芳[1] 李惠敏[1] 

机构地区：[1]湖南大学生命科学与技术研究院湖南大学生物医学工程中心,湖南长沙410082

出　　处：《中国医师杂志》2006年第9期1189-1192,共4页Journal of Chinese Physician

基　　金：国家重点基础研究发展规划(2002CB513100);国家科技攻关计划资助项目(2003BA310A16);湖南省卫生厅课题(B2003-115)资助项目

摘　　要：目的探讨p53及p21waf1蛋白表达与人舌癌细胞系Tca8113多药耐药性(MDR)的关系。方法采用脂质体介导的转染技术,将野生型p53(wt-p53)基因导入人舌癌耐药细胞系Tca8113/BLM;分子信标(Molecular Beacon)法检测人舌癌细胞系Tca8113及其耐药细胞系Tca8113/BLM中p53基因的mRNA水平;W estern blot检测亲本、耐药细胞和转染细胞中p53、p21waf1、P-gp和MRP蛋白的表达变化;MTT法检测不同细胞的药物敏感性,流式细胞仪检测细胞周期的改变,激光共聚焦显微镜观察细胞凋亡。结果在人舌癌耐药细胞系Tca8113/BLM中p53基因的表达明显下调,并且未检测到p21waf1蛋白的表达;转染了wt-p53的耐药细胞Tca8113/BLM/p53中p53和p21waf1蛋白表达均显著上调,而P-gp和MRP的蛋白表达则明显下调,其对药物BLM的敏感性增加10.1倍(P<0.01);转染细胞的周期也发生改变,G2期细胞增加,G1和S期细胞减少,转染细胞明显出现凋亡。结论人舌癌耐药细胞系Tca8113/BLM多药耐药性的产生,可能通过p53和p21waf1的表达下调使得多药耐药性相关蛋白P-gp和MRP表达上调来实现。Objective To study the relationship between the expression of p53 and its downstream effecter p21^wafl protein and multidrug resistance （MDR） of Tca8113/BLM cell line. Methods The cDNA of wildtype p53 gene was introduced into the drug-resistance cell line Tca8113/BLM by Lipofectamine-mediated transfection. The expression of p53 mRNA in Tca8113/BLM and its parental counterpart Tca8113 cell lines were analyzed using molecular beacons. The expression of p53, p21^wafl , P-gp, and MRP proteins in Tca8113/BLM cell line transfected by wt-p53 cDNA ,the untransfected control Tca8113/BLM cell line, its parental counterpart Tca8113 were analyzed by Western blotting. MTT assay was used to determine the drug-sensitivity of different cell lines. The cell cycle distribution and apotosis of different cell lines were also observed by flow cytometery （ FCM ） and laser confocus microscope （ LCM ）, respectively. Results The expression of p53 protein was significantly lower, and no p21^wafl protein was detected in Tca8113/BLM cells. The protein expression of P-gp and MRP were significantly higher in Tca8113/BLM cells than in Tca8113 cells. By Western Blotting, the expression of p53 and p21^wafl proteins was significantly increased, while the expression of P-gp and MRP proteins was significantly decreased in Tca8113/BLM/p53 cells. Comparing to Tca8113/BLM cells, the sensitivity of BLM treatment in Tca8113/BLM/p53 cells was 10. 1 fold increased. Tca8113/BLM/p53 cells decreased at G1 and S phase, and increased at G2 phase. LCM results showed that the apoptosis of Tca8113/BLM/p53 cells was more obvious than Tca8113/BLM cells when treated with the drug BLM. Conclusion The multi-drug resistance in Tca8113/BLM cell line is involved in down-regulation of p53 and p21^wafl protein levels, and up-regulation of P-gp and MRP protein levels.

关 键 词：舌肿瘤 抗药性 肿瘤 蛋白质P53 癌基因蛋白质p21(ras) 

分 类 号：R739.8[医药卫生—肿瘤]