中文摘要：

目的探讨趋化因子受体CXCR4在肾癌索拉非尼治疗耐药中的作用及可能的机制。 方法2014年1月至2016年6月将肾癌细胞786－O接种于裸鼠前肋侧皮下荷瘤，每点5×106个细胞，在移植瘤体积约100 mm3时，将裸鼠分为对照组和索拉非尼组，每组3只。索拉非尼组予索拉非尼（80 mg/kg，1次/d）灌胃，对照组予等量生理盐水灌胃。对照组第5周肿瘤体积即超过1 500mm3，切取肿瘤作为对照组织；索拉非尼组第8周肿瘤开始加速生长，至第13周肿瘤体积超过1 500mm3，取第13周肿瘤作为耐药组织。采用实时定量PCR、蛋白质印迹和免疫组化法检测对照组织和耐药组织中CXCR4的表达量。构建pcDNA3.1－CXCR4质粒，将其转染至786－O细胞中过表达CXCR4，实时定量PCR和蛋白质印迹法测定过表达效果。使用CCK－8和单克隆形成实验检测细胞的药物反应性变化，比较对照组、过表达组和抑制剂组（CXCR4过表达＋CXCR4抑制剂AMD3100）的耐药性。蛋白质印迹法检测对照组、单用索拉非尼组、过表达CXCR4＋索拉非尼组及过表达CXCR4＋索拉非尼＋AMD3100组786－O细胞存活关键分子PKB、ERK、STAT3的磷酸化情况。 结果与对照组织相比，耐药组织CXCR4的mRNA水平升高（3.22±0.23）倍，蛋白质迹法检测结果显示蛋白表达量平均升高（2.33±0.47）倍，差异均有统计学意义（均P〈0.01）。过表达CXCR4后，CXCR4的mRNA表达量升高了（78.3±5.3）倍，蛋白表达量升高了（2.80±0.95）倍，差异均有统计学意义（均P〈0.01），过表达模型构建成功。用CCK－8法测定对照组、过表达组和抑制剂组对索拉非尼的药物反应性曲线，计算出IC50分别为（7.5±0.8）、（10.3±0.7）、（5.7±0.6）μmol/L，差异有统计学意义（P〈0.05）。在索拉非尼7.5 μmol/L作用下，对照组、过表达组和抑制剂组形成的克隆数分别为（26±5）、（56±12）、（42±9）个，差?

英文摘要：

ObjectiveTo investigate the role and possible mechanism of Chemokine receptor CXCR4 in the drug resistance of sorafenib in renal cell carcinoma. Methods786-O cells were inoculated into the anterior sciatic region of nude mice subcutaneously, 5×106 cells per point. The mice were given normal saline and sorafenib intragastric （80 mg/kg, 1 time/day） when the transplanted tumor volume reached about 100 mm3. The tumor volume in the saline group was more than 1 500 mm3 at the 5th week, and the tumor was taken as the control tissue. Sorafenib group tumors started to grow accelerately at week 8, and the tumor volume was more than 1 500 mm3 at week 13. The 13th week tumors were used as resistant tissue. The expression of CXCR4 in control tissues and drug resistant tissues was detected by real-time quantitative PCR, western blotting and immunohistochemistry. The pcDNA3.1-CXCR4 plasmid was constructed and transfected into 786-O cells. The expression of CXCR4 was detected by real-time quantitative PCR and western blotting. The drug reactivity of the cells was measured by CCK-8 and monoclonal assay to compare the drug resistance of the control group, CXCR4 overexpression group and CXCR4 overexpression ＋ CXCR4 inhibitor AMD3100 group. The phosphorylation of PKB, ERK and STAT3 in the control group, the sorafenib alone group, the overexpressing CXCR4＋ sorafenib group and the overexpressing CXCR4＋ sorafenib＋ AMD3100 group were determined by Western blotting. ResultsCompared with the control tissues, the mRNA levels of CXCR4 in the drug-resistant tissues increased （3.22±0.23） times, and the levels of protein expression increased （2.33±0.47） according to western blotting, the differences were statistically significant （P〈0.01）. After overexpression of CXCR4, the mRNA expression of CXCR4 increased （78.3±5.3） times, and the protein expression level increased （2.80±0.95） times, and the differences were statistically significant （P〈0.01）, indicating that the expression model was establ