基于气相色谱-质谱的代谢组学技术研究紫杉醇诱导的卵巢癌A2780耐药株的生物标志物

Metabolomic profiling based on gas chromatography-mass spectrometer(GC-MS) for identification of potential biomarkers in paclitaxel-resistant A2780 ovarian cancer cell

采用气相色谱-质谱(GC-MS)的代谢组学技术研究人卵巢癌紫杉醇耐药细胞A2780/Taxol与敏感细胞之间内源性代谢物的差异,初步探讨A2780/Taxol细胞生物学特性和耐药机制。使用浓度梯度法建立A2780/Taxol细胞,并通过细胞形态观察,生长曲线测定,药物敏感试验,细胞内多药耐药相关蛋白(MRP1)、P-糖蛋白(P-gp)和肺耐药蛋白(LRP)的水平测定,评价A2780/Taxol细胞的生物学特性。同时,采用GC-MS的代谢组学方法分别对卵巢癌敏感细胞株与紫杉醇耐药细胞株的代谢物指纹图谱进行分析,并通过偏最小二乘法判别分析(PLS-DA)对代谢组学数据进行多元处理。结果表明:人卵巢癌细胞敏感细胞与耐药细胞相比,其中琥珀酸、天冬氨酸、果糖、肌醇等13种内源性代谢物存在显著性差异。糖降解、三羧酸(TCA)循环、肌苷代谢途径发生了异常改变,这些差异改变可能与卵巢癌细胞的耐药机制有关。

Gas chromatography-mass spectrometer （GC-MS） metabolomic was used to identify the differences of potential metabolic between human ovarian cancer paclitaxel-resistant cells （A2780/Taxol） and sensitive ones. Meanwhile, it was used to research biological characteristics and resistance mechanisms of paclitaxel-resistant cells. Paclitaxel-resistant cells were induced by continuous stepwise selection with increasing concentrations of taxol. The biological characteristics were identified by evaluating cell morphology, growth curve, drug susceptibility testing, and intracellular protein expression measurement of multidrug-associated protein （ MRP 1 ）, P-glycoprotein （P-gp） and lung resistance protein （ LRP ）. GC- MS metabolomics was used to investigate endogenous metabolic changes in A2780 paclitaxel-resistant cells.The metabolic differences of paclitaxel-resistant cells were compared by sensitive cells using partial least squares-discriminant analysis （PLS-DA） score plot analysis. Results showed that significant metabolite differences were also observed between human ovarian cancer paclitaxel-resistant cells and sensitive ones. 13 characteristic metabolites, such as succinic acid, aspartic acid, fructose, and inositol et al were identified. Abnormal alterations in metabolic pathways were identified, namely, the glycolysis, tricarboxylic acid （TCA） cycle, inosine metabolism, ovarian cancer cells.