TLR9表达与胃癌临床病理特征关系的初步研究

Relationship between TLR9 and Clinicopathologic Features in Patients with Gastric Cancer

目的观察TLR9在胃癌中的表达及其与胃癌临床病理特征的关系。方法运用Western-blot方法测定胃癌细胞系MGC803 TLR9的表达;用不同浓度的TLR9激动剂CpG ODN(2、5、10μg.mL-1)分别作用于胃癌细胞系MGC803不同时间(24、48、72h)后,用四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞生长抑制率;免疫组化SP法检测30例胃癌组织和15例癌旁组织中TLR9的表达,并分析其与患者临床病理特征的关系。结果 Western-blot法检测到TLR9在胃癌细胞系MGC803中有表达;2μg.mL-1和5μg.mL-1 CpGODN仅在作用的72h有抑制胃癌细胞增殖的作用(P<0.01),而10μg.mL-1 CpG ODN作用24、48h和72h均能显著抑制该细胞的增殖(P<0.01);免疫组化结果显示,在胃癌组织和癌旁组织中均有TLR9的表达,且癌旁组织TLR9的表达高于胃癌组织(P<0.05);高、中分化组TLR9的表达高于低分化组(P<0.05);未观察到TLR9蛋白的表达与患者的性别、年龄、肿瘤大小、部位、TNM分期和淋巴结转移具有相关性。结论 TLR9在胃癌中有表达,且与肿瘤分化程度有关。

Objective To investigate the expression of TLR9 in gastric cancer and the Correlation of TLR9 to clinicopathologic features of gastric cancer. Methods expression of TLR9 in the gastric cancer cells MGC803 were measured by Western - blot. inhibitive effect of TLR9 agonist （ CpG ODN ） 2μg.mL-1 mL -1, 10μg.mL-1in MGC803 cell at 24 h,48 h,72 h were observed by MTT assay. Immunohistochemical SP method was used to detect the expression of TLR9 in 54 cases with gastric cancer and 15 cases of paracancer- ous normal gastric tissues. Results Western - blot showed the expression of TLR9 in gastric cancer cellline MGC803. 2 μg.mL-1 and 5μg.mL-1 CpG ODN inhibited significantly cell proliferation at 72h （P 〈 0.01）. 10μg.mL-1 CpG ODN could significantly inhibit the cell growth at 24h, 48h and 72h （ P 〈 0.01 ）. The expression of TLR9 was significantly higher in the paracancerous tissues than those in the gastric cancer （P 〈 0.05 ）. The expression of TLR9 was significantly higher in the highly - moderately differentiation tissues than those in thepoorly （P 〈 0, 05 ）, however, correlation was not found among TLR9 protein expression and age, gender, tumor size, location, TNM stage or lymphatic metastasis stastus of patients. Conclusion TLR9 can express in tissue of gastric cancer and it was related with the tumor differentiation.