平阳霉素诱导的人舌癌多药耐药相关蛋白质表达谱的建立

Expression profiles for multidrug resistance-associated proteins induced by Pingyangmycin in human tongue cancer

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 the expression profiles of multidrug resistance （MDR）-associated proteins induced by pingyangmycin （PYM） in human tongue cancer. Methods： A model PYM-resistant Tca8113/PYM cell line was developed by stepwise selection （described in a previous report from our laboratory）. It, compared with its parental cell line Tca8113, allows us to evaluate multi-drug resistance using a methyl thiazolyltetrazolium （MTT） assay. Comparative proteomics was used to identify the differential expression of proteins between Tca8113/ PYM and Tca8113 cells. The differential expression levels of the selected proteins were determined by Western blot, and expression of the corresponding mRNAs by real-time, quantitative RT-PCR. Results： The resistance to PYM in Tca8113/PYM cells is 55 times greater than that in Tca8113 cells. Tca8113/PYM cells also showed cross-resistance to cis-diaminedichloro- platinum （cDDP）, Taxol, mitomycin C （MMC）, theprubicin （THP） and adriamycin （ADM）. A total of 18 were identified proteins whose expression differed between the Tca8113/ PYM and Tca8113 lines. Of these, 13 proteins were up-regulated in Tca8113/PYM, while 5 proteins were downregulated. The differential expression of these proteins （as measured by Western blot） were consistent with their corresponding mRNA expression, as measured by real-time quantitative RT-PCR. Conclusion： A protein expression profile of proteins related to PYM resistance in human tongue cancer has been established. These results will indubitably provide new clues to further study the mechanisms of multidrug resistance in human tongue cancer.