T细胞免疫球蛋白和黏蛋白-1在非小细胞肺癌组织的表达及其临床意义

T-cell immunoglobulin and mucin domain 1 expression in human non-small-cell lung cancer tissues and its clinical significance

目的观察T细胞免疫球蛋白和黏蛋白-1(TIM-1)在非小细胞肺癌(NSCLC)癌组织及癌旁组织中的表达,分析其表达水平与NSCLC患者临床病理特征及预后的关系。探讨TIM-1干预对NSCLC细胞株生物学功能的调节作用。方法采用免疫组织化学染色法检测肺癌组织芯片中68例肺鳞状细胞癌、85例肺腺癌和对应癌旁组织中TIM-1的表达,采用秩和检验比较肺癌及癌旁组织中TIM-1表达水平的差异,χ2检验分析肺癌组织中TIM-1表达水平与患者临床病理特征的关系,采用Kaplan-Meier生存分析法分析TIM-1表达水平与患者总生存期关系,拟合Cox模型评价不同指标与患者预后的关系。选用NSCLC细胞株A549和SK-MES-1,经RNA干扰(RNAi)技术构建TIM-1稳定下调表达细胞株,通过细胞计数试剂盒(CCK-8)细胞增殖实验、划痕实验及Transwell实验探讨TIM-1干预对上述肺癌细胞株生物学功能的调节作用,应用GraphPad Prism 5.0软件统计分析。结果在肺癌组织中,TIM-1主要定位于肺癌细胞的胞膜及胞质。TNM分期为Ⅲ+Ⅳ的肺鳞状细胞癌患者与TNM分期为Ⅰ+Ⅱ的肺鳞状细胞癌患者其TIM-1表达比率分别为33.3%和13.2%,差异有统计学意义(χ2=3.969,P<0.05)。经过RNAi及慢病毒转染,成功构建TIM-1稳定下调表达细胞株LV-TIM-1-shRNA及对照组LV-NC。CCK-8实验表明,TIM-敲低表达后细胞增值率显著降低;划痕实验显示LV-TIM-1-shRNA组细胞迁移能力显著低于LV-NC组(A549,24 h:t=17.540,P<0.01;SK-MES-1,24 h:t=11.450,P<0.01);Transwell侵袭实验表明,敲低TIM-1表达后细胞侵袭数量显著减少。结论TIM-1在NSCLC组织中高表达,其表达水平与肺鳞状细胞癌患者TNM分期密切相关,TIM-1高表达可作为肺腺癌患者预后评判的独立风险因素,TIM-1在肺鳞状细胞癌组织中的高表达趋势对预测患者预后具有重要意义。

Objective To investigate the expression of T-cell immunoglobulin and mucin domain 1(TIM-1)in non-small-cell lung cancer(NSCLC)tissues and to analyze its clinical significance.To further explore the regulatory effect of TIM-1 intervention on the biological function of NSCLC cell lines.Methods The immunohistochemistry assay and the tissue micro-array were used to examine the TIM-1 expression in lung cancer tissues as well as adjacent normal lung tissues.The correlation between the expression level of TIM-1 and patients’clinic pathological parameters was evaluated by using chi-square test.The lung cancer cell lines A549 and SK-MES-1 were used to construct the stable knockdown expression of TIM-1 using RNA interference(RNAi)method.The cell counting kit-8(CCK-8),wound healing,and Transwell were used to examine the cellular function after TIM-1 knockdown of these cells.Results Higher TIM-1 expression in lung squamous cell carcinoma tissues is significantly correlated with advanced TNM stage(Ⅲ+Ⅳ:33.3%,Ⅰ+Ⅱ:13.2%)(χ2=3.969,P<0.05),but not any other parameters.CCK-8 assay showed that,knockdown expression of TIM-1 make the cell proliferation rate decreased significantly.The wound healing assay showed that the cell migration abilities of LV-TIM-1-shRNA group was significantly lower than LV-NC group(A549,24 h:t=17.540,P<0.01;SK-MES-1,24 h:t=11.450,P<0.01).The Transwell assay showed that the cell invasion abilities was significantly decreased after knockdown expression of TIM-1.Conclusion Our results showed that the TIM-1 was highly expressed in non-small-cell lung cancer tissues,and its expression level was significantly correlated with TNM stage in squamous cell carcinoma patients.TIM-1 expression in lung adenocarcinoma tissues could be used as independent prognostic predictor for the patients,while higher TIM-1 expression in lung squamous cell carcinoma tissues trends significant for the prognostic prediction for the patients.Knockdown expression of TIM-1 could inhibit the cell viability,and the abilities of migration and invasion,and thus TIM-1 could be used as potential therapeutic target for lung cancer.