Xie Xiao-Que, Zhao Qi-Hong, Wang Hua, Gu Kang-Sheng
Xiao-Que Xie, Hua Wang, Kang-Sheng Gu, Department of Oncology, the First Affiliated Hospital of Anhui Medical University, Hefei 230032, Anhui Province, China.
World J Gastroenterol. 2017 Feb 21;23(7):1189-1202. doi: 10.3748/wjg.v23.i7.1189.
To explore novel therapeutic target of cisplatin resistance in human gastric cancer.
The sensitivity of SGC7901 cells and cisplatin-resistant SGC7901 cells (SGC7901/DDP) for cisplatin were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. High-quality total RNA which isolated from SGC7901/DDP cells and SGC7901 cells were used for mRNA microarray analysis. Results were analyzed bioinformatically to predict their roles in the development of cisplatin resistance and the expression of 13 dysregulated mRNAs we selected were validated by quantitative real-time polymerase chain reaction (qRT-PCR).
SGC7901/DDP cells highly resistant to cisplatin demonstrated by MTT assay. A total of 1308 mRNAs (578 upregulated and 730 downregulated) were differentially expressed (fold change ≥ 2 and -value < 0.05) in the SGC7901/DDP cells compared with SGC7901 cells. The expression of mRNAs detected by qRT-PCR were consistent with the microarray results. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes pathway and protein-protein interaction analysis demonstrated that the differentially expressed mRNAs were enriched in , , , , , signaling pathways which may be involved in cisplatin resistance. Several genes such as , , , , and may associated with drug resistance of gastric cancer cells to cisplatin.
Exploration of those altered mRNAs may provide more promising strategy in diagnosis and therapy for gastric cancer with cisplatin resistance.
探索人胃癌顺铂耐药的新治疗靶点。
采用3-(4,5-二甲基噻唑-2)-2,5-二苯基溴化四氮唑蓝（MTT）法检测SGC7901细胞和顺铂耐药的SGC7901细胞（SGC7901/DDP）对顺铂的敏感性。从SGC7901/DDP细胞和SGC7901细胞中分离出的高质量总RNA用于mRNA微阵列分析。对结果进行生物信息学分析，以预测它们在顺铂耐药发生中的作用，并通过定量实时聚合酶链反应（qRT-PCR）验证我们选择的13个失调mRNA的表达。
MTT法显示SGC7901/DDP细胞对顺铂高度耐药。与SGC7901细胞相比，SGC7901/DDP细胞中共有1308个mRNA（578个上调和730个下调）差异表达（倍数变化≥2且P值<0.05）。qRT-PCR检测的mRNA表达与微阵列结果一致。基因本体论、京都基因与基因组百科全书通路和蛋白质-蛋白质相互作用分析表明，差异表达的mRNA富集于可能参与顺铂耐药的细胞增殖、细胞周期、凋亡、DNA修复、PI3K-Akt、MAPK信号通路。一些基因如ABCB1、ABCC1、ABCC2、ABCG2、MDR1和MRP1可能与胃癌细胞对顺铂的耐药性有关。
探索这些改变的mRNA可能为顺铂耐药的胃癌诊断和治疗提供更有前景的策略。
