上皮性卵巢癌患者肿瘤局部微环境中Treg/Th17比例平衡的研究

The imbalances between the Treg cell and Th17 cell in the tumor microenvironment of the patient with epithelial ovarian cancer

目的:探讨上皮性卵巢癌(EOC)患者外周血、卵巢癌组织和癌旁腹膜中Treg/Th17是否存在失衡。方法:选取2011年9月至2013年12月同济大学第一妇婴保健院收治的16例EOC(EOC组)、11例良性上皮性肿瘤(良性肿瘤组)及14例健康成年女性(对照组),收集患者外周血并分离淋巴细胞;收集4例EOC患者腹水并分离淋巴细胞。流式细胞术检测外周血及腹水中Th17、Treg细胞占CD4+T细胞的比例。选取同一时间段在本院手术患者术中留取的卵巢肿瘤组织、腹膜组织及转移灶组织,包括13例EOC、16例EOC种植灶癌旁腹膜、5例良性卵巢肿瘤组织及腹膜,用免疫荧光染色分析Th17及Treg细胞在良性卵巢肿瘤、EOC原发灶和癌旁腹膜中的浸润情况。结果:(1)EOC患者外周血中Treg比例为(5.16±3.85)%,显著高于对照组[(2.41±1.76)%])和良性肿瘤组[(2.3873±2.336)%](P=0.025,P=0.043),后两组比较无统计学差异。EOC患者外周血中Th17细胞比例为(3.15±3.045)%,显著高于对照组[(1.22±1.13)%](P=0.044);良性肿瘤患者[(1.93±1.745)%]与EOC患者和对照组比较,差异均无统计学意义;Treg/Th17细胞比值在3组间均无统计学差异。EOC患者腹水与外周血中的Th17、Treg细胞比例及Treg/Th17比值比较,差异均无统计学意义(P>0.05)。(2)EOC组肿瘤组织中Treg和Th17细胞比例及Treg/Th17比值分别为(0.1062±0.077)%、(0.143±0.056)%和0.80±0.56,与良性肿瘤组织[0%、(0.0789±0.11)%、0]比较,均显著增高(P均<0.05)。(3)卵巢良性肿瘤腹膜中未见Treg及Th17细胞浸润,EOC腹膜种植灶癌旁腹膜中Treg、Th17细胞比例及Treg/Th17比值分别为(0.1024±0.1)%、(0.2254±0.23)%和0.8113±1.097,较良性肿瘤组均显著提高(P<0.05)。(4)早期和晚期EOC组织中Th17、Treg比例以及Treg/Th17比值比较,差异均无统计学意义(P>0.05)。结论:卵巢癌患者外周血Treg和Th17比例升高,但并未发现失衡。卵巢癌组织及癌旁腹膜微环境中存在失衡,这一失衡可能促进肿瘤增殖与迁移。

Objective：To investigate the balance of Treg / Th17 in the peripheral blood, tumor tissue and peritoneum of epithelial ovarian cancer. Methods：We collected the peripheral blood from 14 healthy donors（control group）,11 benign ovarian tumor patient（benign group） and 16 EOC patients（EOC group） and also collected 4 cases ascites from EOC patients during Sep. 2011 to Dec. 2013 in Shanghai First Maternity and Infant Hospital. Lymphocyte in periph-eral blood and ascites were isolated,and we used Flow cytometry to detect the rate of Th17 cell and Treg cell in CD4＋ T lymphocyte. Collect the tissue samples after the operation during the same period,including EOC tissues （13 cases）,peritoneum tissue adjacent to carcinoma （16 cases）,benign tumor tissues and normal peritoneum （5 cases）,The infiltration of Th17 cell and Treg cell in tumor tissue and peritoneum were detected by immunofluorescence. Results：（1） The frequency of Treg cell and Th17 cell in EOC patients[（5. 16±3. 85）% ] was significantly higher than that in healthy donors[（2. 41 ±1. 76）% ] （P = 0. 025） and patients with benign tumors[（2. 3873±2. 336）% ] （P = 0. 043）. There was no significance difference between the patients with benign tumors and healthy donors. The frequency of Th17 cell in EOC patients[（3. 15±3. 045）% ]was significantly higher than that in healthy donors[（1. 22±1. 13）% ]（P= 0. 044）. There was no significance difference between the patients with benign tumors[（1. 93±1. 745）% ] and EOC patients. Neither was between the patients with benign tumors and healthy donors. Ratio of Treg / Th17 cell in three gorups shows no significance difference. （2） Compared with the patient with benign tumors[Treg cell 0% ,Th17 cell 0. 789% ,Treg / Th17 0% ],the expression of those two cells was significant higher in EOC patients＇ ovarian tumor（Treg cells was[（0. 1062±0. 077）% ],Th17 was[（ 0. 143±0. 056）% ], the ratio of Treg /Th17 is 0. 80±0. 56 ） （P〈0. 05）. （3） In peritoneum of EOC patient,the frequency of Treg cells was[（ 0. 1024±0. 1）% ],（P〈0. 05）,Th17 was[（0. 2254±0. 23）% ]（P〈0. 05）,the ra-tio of Treg / Th17 was 0. 8113±1. 097 （P〈0. 05）. Both cells and the ratio of Treg / Th17 were significantly higher than that in benign peritoneum,whose Treg cell and Th17 cell wasn＇t ob-served. （4） Th17,Treg and ratio of Treg / Th17 cell in tumor tissue and peritoneum shows no significance difference between stage I,II and stage III/ IV patient. Conclusion：Though the fre-quency of Treg cell and Th17 cell of EOC patient was significantly higher,the balance of these two cells wasn＇t disturb. While the imbalance between Treg / Th17 in EOC tumor tissue and per-itoneum was observed in our study ,indicating the imbalance between those two cells in the tumor microenvironment may contribute to EOC invasionand migration.