【作者】 卿翔；

【导师】 谭国林；

【作者基本信息】 中南大学 ， 临床医学， 2013， 博士

【摘要】 目的：探索TXR1基因在鼻咽癌紫杉醇耐药中的作用,利用RNA干扰技术抑制鼻咽癌紫杉醇耐药细胞CNE-1/Taxol的TXR1基因表达,同时研究TXR1基因的表达与鼻咽癌紫杉醇耐药的相关性。方法：1.通过脂质体介导转染siRNA至鼻咽癌紫杉醇耐药细胞株CNE-1/Taxol。2.利用实时荧光定量RT-PCR. Western-blot方法分别检测RNA干扰前、后的鼻咽癌紫杉醇耐药CNE-1/Taxol细胞株的TXR1基因表达差异。3.利用MTT实验检测TXR1基因表达抑制后的鼻咽癌紫杉醇耐药细胞CNE-1/Taxol的耐药性变化,观察TXR1基因在紫杉醇耐药过程中的作用。结果：1.针对TXR1基因的siRNA转染耐药细胞CNE-1/Taxol后,耐药细胞TXR1的mRNA及蛋白表达均明显下调。干扰组和阴性对照组的mRNA表达分别是脂质体组mRNA表达量的0.12±0.02倍、0.97±0.07倍,干扰组分别和阴性对照组、脂质体组比较,差异具有统计学意义(P<0.01)。干扰组、阴性对照组的蛋白表达量分别是脂质体组的0.36±0.07、1.02±0.03倍,干扰组与脂质体组、阴性对照组的蛋白表达相比较,差异有统计学意义(P<0.01)。2.TXRl基因表达抑制后,CNE-1/Taxol对紫杉醇的敏感性显著增高,其IC50值降低了2.12倍,从而部分逆转CNE-1/Taxol耐药。结论：1.特异性siRNA分子能有效的抑制鼻咽癌紫杉醇耐药细胞的TXR1基因表达。2.运用RNA干扰技术抑制TXR1基因可能部分逆转人鼻咽癌紫杉醇耐药细胞CNE-1/Taxol的耐药性。TXR1基因可能成为逆转鼻咽癌细胞紫杉醇耐药及肿瘤治疗的靶分子。图7幅,表2个,参考文献53篇更多还原

【Abstract】 Objectives:To explore the role of TXR1played in taxol resistance of nasopharyngeal carcinoma, the expression of TXR1genes was kocked down by RNA interference in paclitaxel resisitant cells of nasopharyngeal carcinoma, and the correlation was analyzed between the expression of TXR1genes and paclitaxel resistance.Methods:1.TXR1siRNA was transfected into the NPC CNE-1/Taxol cells by using liposomes2000.2. The expressional change of both TXR1mRNA and protein was detected by real time RT-PCR and western blot, respectively.3. Proliferation inhibition rate of CNE-1/Taxol cells was assayed by MTT in drug-resistant NPC cells.Results:1. The TXR1-targeted siRNA can effectively decrease the expression level of TXR1gene mRNA and protein in NPC cells.2. The sensitivity to paclitaxel of CNE-1/Taxol was significantly increased after inhibition of TXR1gene expression by siRNA technology, and a2.12-fold decease was observed in its IC50value.Conclusion:1. The siRNA can effectively decrease the mRNA and protein expression level of TXR1gene in nasopharyngeal carcinoma cells, and increase the sensitivity to paclitaxel in CNE-1/Taxol, and reverse the drug resistance.2. The results indicated that the TXR1gene is one of the important target molecules in the reversion of the paclitaxel-resistance in nasopharyngeal carcinoma cells.更多还原