肾腺癌中T细胞亚群和HSP96肿瘤抗原肽复合物关系的探讨

摘要

Objective: To investigate the distribution of T cell subsets and its significance in renal adenocarcinoma, to explore the differences in the expression of heat shock protein 96 ( HSP96 ) between renal adenocarcinoma with necrosis and that without necrosis, and to explore the tumor-associated antigen peptides by extracting HSP96 peptide complex from the renal adenocarcinoma tissue.Methods: Samples from 30 cases of renal adenocarcinoma with hemorrhagic necrosis, 17 cases of renal adenocarcinoma without hemorrhagic necrosis and 13 patients treated with arterial embolization were detected for distribution of T subsets and HSP96 expression by two-step immunohistochemistry.Heat shock protein peptide complex 96 was extracted by the classical method and was analyzed by mass spectrometry.Results: The differences in CD3+ T cells, CD4 +T cells, CD8 + T cells and CD4 + / CD8+ ratio were significant between the two groups ( P ＜ 0.05 ).No significant difference was found in HSP 96 expression between the two groups ( P ＞ 0.05 ).The LC-ESI-MS-MS found LVPLEGWGGNVMand PPVYYVPYVVL as the two major peptide sequences, with molecular weight of 1270.68 and 1307.70.Conclusion:Preoperative arterial embolization and hemorrhagic necrosis can increase the number of T cells in local renal adenocarcinoma, which is of great importance for killing cancer cells and inhibiting spread of cancer cells.The characteristics of gp96 binding peptides showed that they may be tumor-associated antigen peptides, but furhter verification is warranted.%目的:探讨肾腺癌无出血坏死者和肾腺癌有出血坏死与行肾动脉栓塞术者T细胞亚群的分布及意义、两组肾腺癌患者肿瘤组织中热休克蛋白(HSP96)的分布有无差异及意义.通过提取出肾腺癌患者癌组织中的HSP96肽复合物找到肿瘤相关的抗原肽.方法:1)选取肾腺癌无出血坏死患者30例,肾腺癌有出血坏死者17例,行肾动脉栓塞术患者13例.应用两步法免疫组织化学染色观察其T亚群的分布及HSP96的含量和分布.2)通过经典的HSP96-抗原肽复合物提取方法提取出热休克蛋白肽复合物并进行质谱分析.结果:1)两组比较CD3+T细胞、CD4+T细胞、CD8+T细胞及CD4+/CD8+的比值有统计学差异(P<0.05).2)两组比较肾腺癌中含有的HSP96没有统计学意义(P>0.05).3)经LC-ESI-MS-MS测得其中两个主要肽段的序列分别为LVPLEGWGGNVM和PPVYYVPYVVL,其分子量分别为1 270.68和1 307.70.结论:术前行动脉栓塞术和肾腺癌有出血坏死者因为肿瘤细胞的坏死而使其细胞内具有抗原性的肽段释放出来,从而提高了效应T细胞在肿瘤局部的数量,对肿瘤组织的杀伤及抑制肿瘤细胞的扩散具有重要的意义.根据gp96结合肽的特点推测其可能为肿瘤抗原肽,有待进一步鉴定.