文献类型：期刊文献

中文题名：长春新碱耐药结肠癌细胞株HCT-8／VCR的建立及其耐药机制的初步研究

英文题名：Preliminary study on establishment of a VCR-resistant colon carcinoma cell line and its underlying mechanism of drug-resistance

作者：李长龙[1,2];刘建文[1];萨晓婴[2];邓皖利[3];许建华[3];范忠泽[3]

机构：[1]华东理工大学药学院药物科学系,上海200237;[2]浙江省医学科学院实验动物中心实验动物与安全性研究重点实验室,杭州310013;[3]上海中医药大学附属普陀医院中医肿瘤科,上海200062

年份：2009

卷号：29

期号：1

起止页码：31

中文期刊名：肿瘤

外文期刊名：Tumor

收录：CSTPCD;;Scopus;北大核心:【北大核心2008】；CSCD:【CSCD2011_2012】；

语种：中文

中文关键词：结肠肿瘤;长春新碱;抗药性,肿瘤;细胞系,肿瘤

外文关键词：Colonic neoplasms; Vincristine; Drug resistance,neoplasm; Cell line,tumor

摘要：目的:建立长春新碱(vincristine,VCR)耐药结肠癌细胞株,初步探讨该细胞株的多药耐药机制。方法:通过逐步增加药物浓度的方法建立VCR耐药结肠癌细胞株。细胞毒实验采用MTT法,细胞周期测定采用碘化丙啶荧光染色法,P-糖蛋白(P-gp)测定采用FCM方法,细胞内VCR浓度测定采用高效液相色谱(high performance liquid chromatography,HPLC)法,多药耐药相关基因(MDR1、MRP1)的表达采用RT-PCR方法检测。结果:HCT-8/VCR的细胞倍增时间和细胞周期与敏感细胞株HCT-8相似。HCT-8/VCR对VCR的耐药倍数分别为19.7,对其他4种化疗药物也表现出不同的抗性。HCT-8/VCR细胞内VCR和DOX的累积量降低(P<0.01),且其细胞表面P-gp表达增加(P<0.01),MDR1 mRNA增加,但MRP1 mRNA没有变化。结论:建立了对VCR产生耐药的结肠癌细胞株HCT-8/VCR,其耐药机制可能为P-gp过量表达导致细胞外排活性增强,预期该细胞株可用于以P-gp为靶点的多药耐药逆转剂的研究。
Objective ： To establish a vincristine （VCR） -resistant colon eareinoma cell line and explore the mechanism for drug resistance. Methods： The VCR-resistant colon carcinoma cell line was established by exposing the cells to the inereasing concentrations of VCR. The MTT assay was used to deteet the eytotoxieity. Cell cycle was deteeted by propidium iodide （PI） fluoreseenee staining. Flow cytometry was used to deteet the P-gp on the surface of the eells. The intracellular eoncentrations of DOX and VCR were detected by fluorospeetrophotometer and high performance liquid chromatography （ HPLC ） , respeetively. Expressions of multidrug resistance gene MDR1 and MRP1 were assessed by RT-PCR. Results ： The cell double time and the cell eycle of HCT-8/VCR cells were similar to that of sensitive HCT-8 cells. The resistance of HCT8/VCR cells to VCR was 19.7 fold of HCT-8 cells. HCT-8/VCR displayed different resistance to other four antieaneer drugs. The intraeellular concentrations of DOX and VCR in HCT-8/VCR cells were significantly decreased （P 〈 0.01 ）. The expressions of P-gp and MDR1 mRNA were significantly increased but the expression of MRP1 had no significant change in HCT-8/VCR cells （P 〈0.01 ）. Conclusion： A VCR-resistant colon cancer HCT-8/VCR cell line was established. The mechanism is related to the overexpression of P-gp thereby causing the increased efflux of exocytosis. We speculate that HCT-8/VCR could be used in the study of P-gp targeted multidrug resistance reversing agents.