紫杉醇耐药人胃癌细胞HGC-27/PTX的建立及其特征鉴定  

Establishment and characterization of paclitaxel-resistant gastric cancer cell HGC-27/PTX

作　　者：李艳艳[1] 陈冬芍 刘镇涛[1] 葛赛[1] 沈琳[1] 高静[1] 

机构地区：[1]北京大学肿瘤医院暨北京市肿瘤防治研究所消化肿瘤内科恶性肿瘤发病机制及转化研究教育部重点实验室,北京100142

出　　处：《基础医学与临床》2017年第8期1146-1151,共6页Basic and Clinical Medicine

摘　　要：目的建立紫杉醇(PTX)耐药人胃癌细胞HGC-27/PTX,探讨耐药前后细胞特征的变化并初步分析紫杉醇耐药机制。方法逐步递增紫杉醇浓度并间歇作用于人胃癌细胞系HGC-27,建立紫杉醇耐药细胞HGC-27/PTX。用CCK-8及流式细胞仪检测细胞的半数抑制浓度(IC50)及细胞周期分布,RNAseq法筛选紫杉醇耐药前后差异表达基因及通路。结果 9个月后,相同浓度紫杉醇对HGC-27/PTX细胞的增殖抑制作用较亲本HGC-27细胞明显减弱(P<0.05),耐药细胞的形态较亲本细胞略有不同。与亲本HGC-27细胞相比,HGC-27/PTX细胞的S期及G2/M期细胞比例明显增多(P<0.01)。HGC-27/PTX细胞较亲本HGC-27细胞具有274个显著差异表达基因(DEGs),表达上调与下调基因分别有130个和144个,差异基因主要富集在ECM-receptor interaction通路(P<0.001)和PI3KAkt信号通路(P<0.05),这为逆转紫杉醇耐药提供有力线索。结论成功建立紫杉醇耐药胃癌细胞HGC-27/PTX并可体外稳定传代,为深入研究耐药机制提供了理想的体外实验模型。Objective To establish the paclitaxel-resistant gastric cancer cell（HGC-27/PTX） and to investigate the changes of characteristics before and after resistance, as well as the possible resistant mechanisms. Methods The paclitaxel-resistant gastric cancer cell HGC-27/PTX was established by increasing paclitaxel dose gradually and in- termittently. The IC50（50% inhibitory concentration） and cell cycle were determined by CCK-8 assay and flow cy- tometry, respectively. The differentially expressed genes （DEGs） and signaling pathways were analyzed using RNAseq. Results The establishment of HGC-27/PTX cells lasted 9 months, and the sensitivity of paclitaxel of HGC-27/PTX cells was significantly lower than parental cells （P〈0. 05 ）. Compared to parental cells, the morphol- ogy of HGC-27/PTX cells was slightly different, and the proportion of S and G2/M phase was obviously increased（ P〈 0. 01 ）. A total of 274 DEGs were identified between the resistant and parental cells with 130 genes up-regulated and 144 genes down-regnlated.DEGs were significantly enriched in extracellular matrix （ECM）-receptor interaction （P〈 0. 001） and PI3K-Akt signaling pathways （P〈0. 05）, which could provide evidences for reversing paclitaxel resist- ance. Conclusions The paclitaxel-resistant gastric cancer cells HGC-27/PTX was established with stable culturein vitro, which provides an ideal model for future study on the mechanism of drug resistance.

关 键 词：胃癌 紫杉醇耐药 RNASEQ 

分 类 号：R735.2[医药卫生—肿瘤]