胃肠道间质瘤Sunitinib耐药细胞株的建立及其生物学特性的研究

Establishment and biologicaI characters of an Sunitinib-resistant cell line of gastrointestinal stromal tumor

目的:构建胃肠道间质瘤耐药细胞模型GIST882-Rs。方法:通过间歇浓度梯度倍增法针对胃肠道间质瘤细胞株GIST882进行体外诱导,使其对Sunitinib靶向药物耐药,研究构建耐药细胞模型GIST882-Rs。使用显微镜对细胞形态进行观察,通过流式细胞仪针对细胞周期进行检测,CCK8法计算细胞半数致死浓度(IC50)及耐药指数。结果:耐药细胞在细胞形态方面具有上皮样特征,核浆比增加;在高倍镜之下能够发现多核细胞以及胞质内部的颗粒样物质有所提升。耐药细胞GIST882-Rs与GIST882相对比而言,针对Sunitinib的IC50有所提升,分别为(127.50±2.15)μmol和(23.98±1.85)μmol,差异有统计学意义(P<0.05),耐药指数为5.30。耐药前后细胞周期变化,S期由52.51%下调至39.87%,G 0/G 1期由25.22%上升至32.21%。结论:获取到耐药细胞GIST882-Rs,GIST882-Rs针对Sunitinib靶向治疗所具备的敏感性显著下调。

Objective To establish the Sunitinib-resistant model GIST882-Rs.Methods The intermittent concentration gradient doubling method was used to get Sunitinib-resistant cell line GIST 882-Rs.Changes of cell morphology were observed and compared under the opticalm icroscopy.Changes of cell cycle were assayed and compared by using flow cytometer.The growth curvature,half maximal inhibitory concentration(IC50)were calculated by using cell counting kit(CCK8)assay.Results Significant changes occurred in the morphologic profiles,such as epithelioid shape,the increased ratio of nucleolus to cytoplasm,emerging multinucleated cel1 and increase of granule sub stances.IC50 for GIST882 and GIST 882-Rs were(127.50±2.15)μmol and(23.98±1.85)μmol,and there were significant differences(P<0.05).The resistance index of GIST882-Rs to Sunitinib was 5.30 fold of GIST882 cells.The cell cycle of GIST882 and GIST882-Rs was respectively 52.51%and 39.87%in S phase 25.22%and 32.32%in G0/G1 phase.Conclusions The sensitivity of GIST 882-Rs to Sunitinib treatment is decreased significantly.