摘要
目的:建立人食管癌紫杉醇耐药细胞系Ec9706/P-1并研究其生物学特征,为食管癌临床治疗提供实验依据。方法:小剂量逐步间歇诱导,建立人食管癌紫杉醇耐药细胞系Ec9706/P-1。用MTT法检测该耐药细胞株的多药耐药性及耐药指数,采用流式细胞术进行细胞周期分析。应用罗丹明(Rh123)排出试验定测P-gp功能。采用免疫组织化学方法检测多药耐药蛋白(MDR1)、多药耐药相关蛋白(MRP1)的表达情况。结果:Ec9706/P-1细胞与Ec9706细胞镜下形态无明显差别,体积稍大;Ec9706/P-1生长缓慢(P<0.05),倍增时间延长(P<0.05),大部分肿瘤细胞集中在G0+G1期(P<0.05);MTT结果显示Ec9706/P-1对PIX的耐药指数为6.68,具有紫杉醇耐药性;Rh123排出试验显示Ec9706/P-1细胞平均荧光强度显著降低;免疫细胞化学方法检测发现Ec9706/P-1多药耐药蛋白(MDR1)表达明显升高(P<0.05),多药耐药相关蛋白(MRP1)表达无统计学意义(P>0.05)。结论:Ec9706/P-1具有明确的耐药性,但耐药蛋白表达不一致,耐药机制复杂,可用于食管癌临床治疗的基础研究。
Objective： To establish a new human esophageal carcinoma cell line that is paclitaxel-resistant （Ec9706/P-1） and to research its biological characteristics and provide evidence for clinical treatment of esophageal carcinoma. Methods： We established the paclitaxel-resitant esophageal carcinoma cell line Ec9706/P-1 by treating the Ec9706 parental esophageal carcinoma cells intermittently with low concentrations of drugs. Muhidrug resistance to anticancer agents was evaluated by MTT assay. Flow cytometry （FCM） was performed to analyze the cell cycle of Ec9706/P-1 and Ec9706. Rhodamine 123 efflux was assessed and the expression of p-glycoprotein was evaluated. Expression of multidrug resistant protein and muhidrug resistance-associated protein was determined by immunohistochemistry. Results： Ec9706/P-1 and Ec9706 were not significantly different microscopically other than a slight difference in volume. Growth of Ec9706/P-1 was slow, with an extended doubling time, and most of the cancer cells were found in stage G0＋ G1 （P〈0.05）. The drug resistance index for Ec9706/ P-1 was 6.68, based on data from the MTT assays. Mean fluorescence intensity in Ec9706/P-1 significantly declined by Rh123 assay. Expression of multidrug resistant protein in Ec9706/P-1 was significantly elevated （P〈0.05）. Expression of multidrug resistance-associated protein was not statistically significant （P〉0.05）. Conclusion： Ec9706/P-1 was confirmed to be multidrug resistant. However, expression of multidrug resistant protein alone did not explain this phenotype and the mechanism seems quite complicated.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2007年第13期731-734,共4页
Chinese Journal of Clinical Oncology
基金
河南省卫生厅创新人才基金资助(编号:2003013)
关键词
食管癌细胞
紫杉醇
耐药
Esophageal carcinoma Paclitaxel Drug resistance
