中文摘要：

目的探讨Toll样受体4（Toll-like receptor 4,TLR4）在人淋巴瘤细胞株中的表达及其对细胞增殖和耐药的影响。方法采用RT-PCR、q PCR和Western blot检测8株淋巴瘤细胞株中TLR4 m RNA及蛋白的表达情况并筛选出TLR4高表达株,对高表达株进行基因测序以排除My D88 L265P基因突变。将TLR4高标达细胞株分为空白对照组、TAK-242组、细菌脂多糖（lipopolysaccharides,LPS）组和LPS＋TAK-242组,分别进行细胞增殖和阿霉素耐药实验。采用CCK-8试剂盒检测其增殖活性和细胞杀伤率,用Western blot法检测增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）和P-糖蛋白（P-glycoprotein,P-gp）的变化。结果 8株淋巴瘤细胞株中TLR4 m RNA和蛋白广泛表达,其中Burkitt淋巴瘤细胞株Raji的表达水平最高。Raji细胞My D88基因为野生型。与空白对照组相比,TAK-242和LPS＋TAK-242组Raji细胞的增殖活性无明显改变（P=2.19,P=1.85）,LPS组Raji细胞的增殖活性明显升高（P=0.016）,LPS组PCNA蛋白表达明显升高（P=0.009）。阿霉素在半数抑制浓度下各组杀伤率分别为：空白对照组（49.23±2.03）%、TAK-242组（51.41±1.12）%、LPS组（24.65±3.17）%、LPS＋TAK-242组（41.17±2.69）%,可见LPS组细胞杀伤率明显降低（P=0.002）。P-gp蛋白表达明显升高（P=0.001）。结论 TLR4分子在淋巴瘤细胞株中广泛表达,尤以Burkitt淋巴瘤细胞株Raji最高,激活TLR4可以促使肿瘤细胞增殖和耐药,其机制可能与PCNA和P-gp的表达上调有关。

英文摘要：

Objective To investigate the expression of Toll-like receptor 4（TLR4） in human lymphoma cell lines and its effect on cell proliferation and drug resistance. Methods Eight lymphoma cell lines were detected by RT-PCR, q PCR and Western blot to select the one with highest m RNA and protein expression of TLR4. Gene sequencing of the high expression cell line was detected to exclude My D88 L265 P gene mutation. Cell proliferation and doxorubicin resistance test of the TLR4 high expression cell line were carried out by blank control group, TAK-242 group, Lipopolysaccharides（LPS） group and LPS＋TAK-242 group, respectively. CCK-8 kit was used to detect the proliferation activity and cell death rate, and Western blot was used to detect the changes of proliferating cell nuclear antigen（PCNA） and P-glycoprotein（P-gp）. Results TLR4 m RNA and protein were expressed in eight lymphoma cell lines, and the Burkitt lymphoma cell line Raji was the highest. My D88 gene in Raji was wild type. Compared with the blank control group, the proliferation of TAK-242 group and LPS＋TAK-242 group wasn＇t increased significantly（P=2.19, P=1.85）; the proliferation of LPS group was increased significantly（P=0.016）. The expression of PCNA protein was significantly increased in the LPS group（P=0.009）. Cell death rate of each group in the half inhibitory concentration of doxorubicin： blank control group（49.23±2.03）%, TAK-242 group（51.41±1.12）%, LPS group（24.65±3.17）%, and LPS＋TAK-242 group（41.17±2.69）%. The cell death rate of LPS group was significantly lower（P=0.002）. The expression of P-gp protein was significantly higher（P=0.001）. Conclusion TLR4 is widely expressed in lymphoma cell lines, especially in Burkitt lymphoma cell line Raji. The activation of TLR4 can induce tumor cell proliferation and drug resistance, and its mechanism may be related to the upregulation of PCNA and P-gp expression.