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目的:建立平阳霉素(pingyangmycin,PYM)诱导的人舌癌多药耐药相关蛋白质表达谱。方法:以前期本室通过用PYM浓度梯度诱导法建立的舌癌PYM多药耐药细胞Tca8113/PYM与其亲本细胞Tca8113为模型,用四甲基偶氮唑盐法检测该耐药细胞株的多药耐药性,运用比较蛋白质组学方法筛选耐药细胞与其亲本细胞中的差异蛋白质表达,并对部分差异蛋白质表达进行实时定量PCR及Western免疫印迹验证。结果:目前Tca8113/PYM细胞对PYM的耐药性约是Tca8113细胞的20倍,同时表现出对顺铂、紫杉醇、丝裂霉素C、四氢吡喃阿霉素和阿霉素存在交叉耐药;比较蛋白质组学筛选了18种差异表达蛋白质,其中在Tca8113/PYM细胞中表达上调的有13种,下调的有5种,实时荧光定量PCR与Western免疫印迹结果与蛋白质组学的结果趋势一致。结论:建立了舌癌PYM耐药相关蛋白质表达谱,为深入研究舌癌多药耐药的分子机制提供了新线索。
Objective: To establish a multidrug resistance-related protein expression profile of human tongue cancer induced by pingyangmycin (PYM). Methods: In our previous study, Tca8113 / PYM and Tca8113 cell lines of PYM were established by PYM concentration-gradient method. The multi-drug resistant cell lines were detected by MTT assay Drug resistance, comparative proteomics method was used to screen the differentially expressed proteins in drug-resistant cells and their parental cells, and some differential proteins were verified by real-time PCR and Western immunoblotting. Results: At present Tca8113 / PYM cells are about 20 times more resistant to PYM than Tca8113 cells and exhibit cross-resistance to cisplatin, paclitaxel, mitomycin C, tetrahydropyraximycin and doxorubicin 18 proteomes were screened by comparative proteomics, of which 13 were upregulated in Tca8113 / PYM cells and 5 were downregulated. The results of real-time fluorescence quantitative PCR and Western blot analysis and proteomics results Consistent. Conclusion: The expression profile of PYM resistance-related protein in tongue cancer was established, which provided new clues for further study on the molecular mechanism of multidrug resistance in tongue cancer.