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.
出处
《肿瘤》
CAS
CSCD
北大核心
2009年第1期31-34,共4页
Tumor
关键词
结肠肿瘤
长春新碱
抗药性
肿瘤
细胞系
肿瘤
Colonic neoplasms
Vincristine
Drug resistance,neoplasm
Cell line,tumor
