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目的:通过对Vav1与肿瘤浸润T淋巴细胞(tumor infiltrating Tlymphocytes,TITL-T)活性关系的研究,提出T细胞失能的可能分子机制;初步探讨TIL-T中Vav1的表达情况及其与肿瘤局部微环境中吲哚胺2,3双加氧酶(indoleamine-2,3-dioxvgenase,IDO)表达的相关性。方法:收集天津医科大学附属肿瘤医院肺外科手术切除的新鲜肺癌标本、癌旁正常肺组织及蜡块40例,通过实时定量RT-PCR检测TIL-T中Vav1 mRNA表达变化;免疫印迹及免疫沉淀技术检测Vav1蛋白表达及磷酸化活性。BrdU法检测T细胞增殖活性此外,运用Real time-PCR法检测肺癌组织中IDO mRNA表达水平,免疫印迹及免疫组化检测IDO蛋白表达情况。结果:部分肺癌局部浸润的T细胞处于功能抑制状态,这种抑制状态可能与细胞内重要的信号传导蛋白Vav1的表达量及活性相关。IDO表达阳性组肺癌标本局部TIL-T中Vav1 mRNA水平及Vav1蛋白的表达和磷酸化水平明显低于IDO表达阴性组(P<0.05)结论:Vav1的表达和活化在TIL-T功能中具有重要的作用。肿瘤局部微环境中的IDO蛋白可能是影响TIL-T中Vav1表达和活化的重要因素之一。IDO可能通过抑制Vav1的表达及磷酸化活化过程,使TIL-T的主动免疫受损,从而降低宿主的抗肿瘤免疫效应。
OBJECTIVE: To investigate the possible molecular mechanism of T cell dysfunction through the study of the relationship between Vav1 and the activity of tumor infiltrating Tlymphocytes (TITL-T). To investigate the expression of Vav1 in TIL-T and its relationship with tumor localization Correlation of indoleamine-2,3-dioxvgenase (IDO) expression in microenvironment. Methods: Forty fresh lung cancer specimens, normal lung tissues and normal paraffin tissues excised from lung surgery at Tumor Hospital of Tianjin Medical University were collected. The expression of Vav1 mRNA in TIL-T was detected by real-time quantitative RT-PCR. Western blotting and immunoprecipitation Technology to detect Vav1 protein expression and phosphorylation activity. In addition, the expression of IDO mRNA in lung cancer tissues was detected by Real time-PCR, and the expression of IDO protein was detected by Western blotting and immunohistochemistry. Results: T lymphocytes partially infiltrated by lung cancer were in a function-inhibited state, which may be related to the expression level and activity of important signaling protein Vav1 in the lung. The expression of Vav1 mRNA and Vav1 protein and phosphorylation in local IDT-positive TIL-T group were significantly lower than those in IDO negative group (P <0.05) .Conclusion: Vav1 expression and activation are important in TIL-T function Role. The IDO protein in tumor microenvironment may be one of the important factors affecting the expression and activation of Vav1 in TIL-T. IDO may reduce the host anti-tumor immune effect by inhibiting Vav1 expression and phosphorylation activation process, thereby impairing the active immunity of TIL-T.