人胃癌顺铂耐药细胞系的建立及其生物学特征

Establishment of a Cisplatin-induced human gastric cancer drug-resistant cell line and its biological characteristics

目的:国内外普遍应用体外建立的耐药细胞系作为研究模型。为探讨胃癌对顺铂耐药的机理,本研究首次建立人胃癌顺铂耐药细胞系并且研究其生物学特性。方法:采用逐步递增顺铂浓度,间歇作用体外诱导法建立人胃癌顺铂耐药细胞系SGC 7901/DDP;M TT法测定药物敏感性;光镜、电镜、M TT法活细胞计数、流式细胞仪及其相关P-糖蛋白的表达等方法观察其生物学特征的改变。结果:历时5个月建成人胃癌顺铂耐药细胞系SGC 7901/DDP,其对顺铂的耐药指数为14.68,并且与5-氟尿嘧啶、丝裂霉素等多种抗癌药有不同程度的交叉耐药性;SGC 7901/DDP的细胞形态及其P-糖蛋白的表达;体外群体倍增时间较亲代细胞延长;细胞周期分析发现其S期与G2/M期细胞减少、G0/G1期细胞增多。结论:SGC 7901/DDP细胞具有耐药特征,耐药性能稳定,与P-糖蛋白的表达有关。

Objective：The drug-resistant cell line established in vitro is generally used as a research model . To explore the mechanism of resistance of gastric cancer to Cisplatin , we designed this study to establish a Cisplatin - induced human gastric cancer drug-resistant cell line and study its biological characteristics. Methods ：A resistant gastric cancer cell line （SGC7901/DDP） was established gradually by increasing dose of cisplatin and intermittent administration; drug sensitivity was detected by MTT assay; the changes of its biological characteristics were determined by using light microscopy, electron microscopy, cell counting by MTT assay, flow cytometry （FCM） , and expression of protein . Results： SGC7901/DDP cell line was developed after 5 months with stable resistance to Cisplatin and the resistance index was 14.68;SGC7901/DDP cells exhibited cross-resistance to many other chemotherapeutic agents （5- Fuorouracil, Mitomycin-C, etc）. The morphology and expression of protein of SGC7901/DDP changed; the doubling time prolonged; and the cell number of S-phase and G2/M- phase decreased, while that of G0/G1 phase increased compared with parent ceils. Conclusion： SGC7901/DDP cell line shows drug- resistant characteristic and drug resistance stability, which is related to the P- glycoprotein expression.