人肺腺癌细胞系SPC-A1多西他赛耐药细胞系的建立及其特性

Establishment and characterization of a docetaxel-resistant variant of human lung adenocarcinoma cell line SPC-A1

背景与目的:多西他赛是近年来应用于临床的一种有效的化疗药物,其独特的作用机制使其具有良好的应用前景,然而,耐药性影响了其临床应用。本研究拟通过体外诱导耐药建立肺癌SPC-A1的多西他赛耐药细胞系,以便于进一步研究其耐药机制及耐药逆转。方法:通过体外逐步增加多西他赛浓度的方法诱导建立肺癌SPC-A1的多西他赛耐药细胞系。MTT比色法检测该耐药细胞的多药耐药特性;流式细胞术检测其细胞周期和细胞内罗丹明蓄积;细胞计数法绘制细胞生长曲线,计算倍增时间;染色体核型分析对比耐药和亲本细胞株在遗传学上的变化。结果:MTT显示SPC-A1/Docetaxel对多西他赛、紫杉醇、顺铂、卡铂、吉西他滨、长春瑞滨、表柔比星、依托泊苷的耐药指数分别为13.20、2.18、1.12、1.39、1.38、0.93、10.14、2.12。流式细胞术检测其细胞周期显示:与亲代细胞比较SPC-A1/Docetaxel处于G1期的细胞增加,S期的细胞减少,G2期的细胞无明显变化。流式细胞仪检测SPC-A1和SPC-A1/Docetaxel细胞内罗丹明蓄积荧光强度分别为1382.26±0.32和806.34±0.49,耐药细胞内罗丹明蓄积较亲本细胞明显减少。SPC-A1和SPC-A1/Docetaxel细胞的倍增时间分别为27.4、35.1h。染色体核型分析两者遗传学无明显变化。结论:经体外诱导耐药建立的SPC-A1/Docetaxel表现出对多西他赛较高的耐药性,并呈多药耐药特性,耐药细胞系SPC-A1/Docetaxel与亲本细胞SPC-A1的生物学特性有明显差异,可用于对多西他赛的耐药机制和逆转耐药研究。

Background and purpose： Docetaxel is one of the most effective chemotherapeutic agents developed in the past few years and has been used for the treatment of various cancers including lung cancer. Drug resistance to docetaxel is one of the main factors accounting for the failure of chemotherapy. This study was done to establish a human lung adenocarcinoma cell line SPC-A1 with the characterization of docetaxel resistance, to investigate its biological mechanism of drug resistance and how to reverse the resistance. Methods： A docetaxel resistant human lung adenocarcinoma cell line SPC-A1/Docetaxel was induced by continuously exposing human lung adenocarcinoma cell line SPC-A1 to gradually increasing doses of docetaxel. The muhidrug resistance of SPC-A1/Docetaxel was evaluated by MTT assay. The distribution of cell cycle and rhodamine 123 accumulation in the two cell lines were detected by flow cytometry. Growth fraction was calculated by cytometry, and differentiation of genetics between drug resistance cell line and its parent cell line was analyzed by karyotype analysis. Results： Index number of drug resistance （IR） of SPC-A1/Docetaxel were 13.20, 2.18, 1.12, 1.39, 1.38, 0.93, 10.14 and 2.12 to docetaxel, paclitaxel, pharmorubicin, cisplatin, carboplatin, gemzar, vinorelbine and etoposide , respectively. Flow cytometry analysis showed G1/G0 arrest in SPC-A1/Docetaxel cell lines and there was no difference in terms of G2 cell proportion between SPC-A1/Docetaxel and SPC-A1 cell lines. Rhodamine concentration in SPC- A1/Docetaxel cells were markedly lower than that in SPC-A1 cells（806.34 ±0.49 Vs. 1382.26 ±0.32）. Cell doubling time of SPC-A1/Docetaxel cells and SPC-A1 cells were 35.1 and 27.4 hours, respectively, with no difference in terms of genetics between the two cell lines. Conclusions： The SPC-A1/Docetaxel cell line showed high resistance to docetaxel, and had the characteristic of multidrug resistance. The cell llne could be useful for the study of drug resistant mechanism and its reversal of docetaxel.