奥沙利铂耐药人结肠癌细胞系的建立及其生物学特性

Establishment and biological characteristics of oxaliplatin-resistant human colon cancer cell lines.

作者信息

Liu Zhen, Qiu Meng, Tang Qiu-Lin, Liu Ming, Lang Nan, Bi Feng

机构信息

Department of Medical Oncology ,West China Hospital, Sichuan University, Chengdu, Sichuan 610041, PR China.

出版信息

Chin J Cancer. 2010 Jul;29(7):661-7. doi: 10.5732/cjc.009.10666.

DOI:10.5732/cjc.009.10666

PMID:20591218

Abstract

BACKGROUND AND OBJECTIVE

Chemotherapy is the main treatment for colon cancer, while multidrug-resistance is the main reason for chemotherapy failure and tumor relapse. This study was to establish two oxaliplatin-resistant colon cancer cell lines and evaluate their biological characteristics.

METHODS

Oxaliplatin-resistant colon cancer cell lines SW620/L-OHP and lovo/L-OHP were established in vitro by continuous exposure to oxaliplatin (L-OHP) of low and gradually increased concentration. Growth curve, cross-resistance and resistance index of the oxaliplatin-resistant cell lines to various anti-cancer agents were determined by CCK8 assay. The expressions of P-glycoprotein (P-gp), multidrug-resistance protein 1 (MRP1) and MRP2 were detected by Western blot. Cell cycle distribution as well as the expression of CD133 and CD44 were measured by flow cytometry.

RESULTS

It took 10 months to establish the SW620/L-OHP and LoVo/L-OHP cell lines with stable resistance to oxaliplatin. Cross-resistance to 5-fluorouracil, etoposide, cisplatin, vincristine and epirubicin but not to paclitaxel was observed. Longer doubling time, higher proportion of cells in G(0)/G(1) phase and lower proportion in G(2)/M phase were observed in the two oxaliplatin-resistant cell lines compared with their parental cell lines. The expression of MRP2 in the oxaliplatin-resistant cells was up-regulated, while those of P-gp and MRP1 had no significant change. CD133 was overexpressed while CD44 level remained unchanged in SW620/L-OHP and LoVo/L-OHP cells.

CONCLUSIONS

SW620/L-OHP and LoVo/L-OHP cell lines show a typical and stably resistant phenotype and may be used as research models.

摘要

背景与目的

化疗是结肠癌的主要治疗方法，而多药耐药是化疗失败和肿瘤复发的主要原因。本研究旨在建立两种耐奥沙利铂的结肠癌细胞系并评估其生物学特性。

方法

通过持续暴露于低浓度并逐渐增加浓度的奥沙利铂（L-OHP）在体外建立耐奥沙利铂的结肠癌细胞系SW620/L-OHP和lovo/L-OHP。采用CCK8法测定耐奥沙利铂细胞系对各种抗癌药物的生长曲线、交叉耐药性和耐药指数。通过蛋白质免疫印迹法检测P-糖蛋白（P-gp）、多药耐药蛋白1（MRP1）和MRP2的表达。采用流式细胞术检测细胞周期分布以及CD133和CD44的表达。

结果

建立对奥沙利铂具有稳定耐药性的SW620/L-OHP和LoVo/L-OHP细胞系耗时10个月。观察到对5-氟尿嘧啶、依托泊苷、顺铂、长春新碱和表柔比星有交叉耐药性，但对紫杉醇无交叉耐药性。与亲本细胞系相比，在两种耐奥沙利铂细胞系中观察到更长的倍增时间、更高比例的G(0)/G(1)期细胞和更低比例的G(2)/M期细胞。耐奥沙利铂细胞中MRP2的表达上调，而P-gp和MRP1的表达无显著变化。在SW620/L-OHP和LoVo/L-OHP细胞中CD133过表达而CD44水平保持不变。