胃癌组织中白细胞介素-6及肿瘤相关巨噬细胞和新生淋巴管的表达及意义

Expressions and correlations of interleukin-6, tumor-associated macrophages and lymphangiogenesis in the tissues of gastric cancer

目的 探讨胃癌组织中IL-6、肿瘤相关巨噬细胞（TAMs）和新生淋巴管的表达的相关性及其与胃癌患者临床病理因素和预后的关系.方法 回顾性分析2012年9月至2013年9月第三军医大学西南医院收治的40例胃癌患者的临床资料.收集患者手术切除的胃组织标本进行研究.采用免疫组织化学染色检测胃癌组织及癌旁组织中IL-6、TAMs特异性抗原CD68、淋巴管特异性蛋白D2-40的表达.分析IL-6、CD68及D2-40间表达的相关性,及其与胃癌患者临床病理因素及预后的关系.采用电话方式进行随访,随访时间截至2014年7月.符合正态分布的计量资料以x^-±s表示,采用t检验,相关性分析采用线性相关分析.IL-6、CD68及D2-40表达与患者临床病理因素的关系采用t检验或方差分析.采用Kaplan-Meier法绘制生存曲线,生存分析采用Log-rank检验.结果 IL-6、CD68和D2-40的表达均位于胃癌组织及癌旁组织细胞的细胞质.胃癌组织中IL-6、CD68、D2-40阳性表达和IL-6与CD68阳性共表达的细胞数分别为（48.0±10.3）个、（26.0±5.5）个、（7.6±3.8）个、（11.4±2.1）个,癌旁组织中其阳性表达的细胞数分别为（11.1±2.3）个、（5.9±1.6）个、（2.5±1.2）个、（2.1±0.7）个,两者上述指标比较,差异均有统计学意义（t=22.021,22.105,8.103,21.893,P＜0.05）.胃癌组织中IL-6阳性表达的细胞数与CD68阳性表达的细胞数、D2-40阳性表达的细胞数及IL-6和CD68阳性共表达的细胞数均呈正相关（r=0.941,0.776,0.781,P＜0.05）.CD68阳性表达的细胞数与D2-40阳性表达的细胞数呈正相关（r=0.840,P＜0.05）.TNM分期为Ⅰ～Ⅱ期的胃癌患者IL-6、CD68、D2-40阳性表达和IL-6与CD68阳性共表达的细胞数分别为（38.6±5.3）个、（21.0±2.2）个、（4.7±1.6）个、（9.7±1.2）个,TNM分期为Ⅲ～Ⅳ期的胃癌患者分别为（57.3±2.6）个、（31.1±1.9）个、（10.6±2.9）个、（13.1±1.3）个;两者上述指标比较,差异均有统计学意义（t=-14.169,-15.509,-8.149,-8.509,P＜0.05）.肿瘤浸润深度为T1～T2期的胃癌患者IL-6、CD68、D2-40阳性表达和IL-6与CD68阳性共表达的细胞数分别为（41.5±12.2）个、（22.2±5.1）个、（5.5±2.6）个、（9.7±1.8）个,肿瘤浸润深度为T3～T4期的胃癌患者分别为（50.1±8.8）个、（27.3±5.1）个、（8.3±3.9）个、（12.0±1.9）个;两者上述指标比较,差异均有统计学意义（t=-2.435,-2.770,-2.212,-3.236,P＜0.05）.无淋巴结转移的胃癌患者IL-6、CD68、D2-40阳性表达和IL-6与CD68阳性共表达的细胞数分别为（39.2±6.7）个、（21.3±3.0）个、（5.2±3.3）个、（9.8±1.4）个,有淋巴结转移的胃癌患者分别为（55.2±6.4）个、（29.9±3.8）个、（9.6±2.9）个、（12.7±1.6）个;两者上述指标比较,差异均有统计学意义（t=-7.722,-7.838,-4.581,-5.756,P＜0.05）.40例患者均获得术后随访,随访时间为6～22个月,中位随访时间为18个月.20例IL-6高表达胃癌患者和20例IL-6低表达胃癌患者的1年累积生存率分别为70.0％、90.0％,两者生存情况比较,差异有统计学意义（χ^2=4.410,P＜0.05）.20例CD68高表达胃癌患者和20例CD68低表达胃癌患者的1年累积生存率分别为70.0％、90.0％,两者生存情况比较,差异有统计学意义（χ^2=4.075,P＜0.05）.20例D2-40高表达胃癌患者和20例D2-40低表达胃癌患者的1年累积生存率分别为65.0％、95.0％,两者生存情况比较,差异有统计学意义（χ^2=4.705,P＜0.05）.20例IL-6和CD68高共表达胃癌患者和20例IL-6和CD68低共表达胃癌患者的1年累积生存率分别为70.0％、90.0％,两者生存情况比较,差异有统计学意义（χ^2=4.152,P＜0.05）.结论 IL-6、TAMs和新生淋巴管在胃癌组织中高表达,IL-6与TAMs、IL-6与新生淋巴管、TAMs与新生淋巴管表达水平均呈正相关,IL-6、TAMs和新生淋巴管与胃癌进展及患者预后密切相关.

Objective To explore the expressions of interleukin-6 （IL-6）,tumor-associated macrophages （TAMs） and lymphangiogenesis in the tissues of gastric cancer,and their correlation with the clinicopathological factors and prognosis of patients.Methods The clinical data of 40 patients with gastric cancer who were admitted to the Southwest Hospital from September 2012 to September 2013 were retrospectively analyzed.The surgical specimens from stomach tissues were collected.The expressions of IL-6,TAMs specific antigen （CD68） and lymphatic vessel specific protein （D2-40） in the tumor tissues and adjacent normal tissues were observed by immunohistochemical double staining technique.The correlations among the expressions of CD68 and D2-40,clinicopathological factors and prognosis of patients.The follow-up was carried out on the patients till July 2014.The measurement data with normal distribution were presented as x^- ± s and analyzed by the t test,and the correlation analysis was conducted.The relationship between the expressions of IL-6,CD68 and D2-40 and the clinicopathological factors was analyzed by the t test and the ANOVA.The survival curve was drawn by the Kaplan-Meier method and the survival analysis was done using the Log-rank test.Results The expressions of IL-6,CD68 and D2-40 were located at the cytoplasm of tumor cells and adjacent normal cells.The number of cells with positive expressions of IL-6,CD68 and D2-40 and positive co-expression of IL-6 and CD68 were 48.0 ± 10.3,26.0 ±5.5,7.6 ±3.8,11.4±2.1 in the tumor tissues and 11.1 ±2.3,5.9 ± 1.6,2.5 ± 1.2,2.1 ±0.7 in the adjacent normal tissues,respectively,showing significant differences （t =22.021,22.105,8.103,21.893,P 〈0.05）.There were significant positive correlations among the number of cells with positive expressions of IL-6,CD68 and D2-40 and positive co-expression of IL-6 and CD68 in the tumor tissues （r =0.941,0.776,0.781,P 〈 0.05）.There was a significant positive correlation in the number of cells with positive expression between CD68 and D2-40 （r =0.840,P 〈 0.05）.The number of cells with positive expressions of IL-6,CD68 and D2-40 and positive co-expression of IL-6 and CD68 in the patients with gastric cancer of stage Ⅰ-Ⅱ were 38.6 ±5.3,21.0 ±2.2,4.7 ± 1.6 and 9.7 ± 1.2,respectively,which were significantly different from 57.3 ± 2.6,31.1 ± 1.9,10.6 ± 2.9 and 13.1 ± 1.3 in the patients with gastric cancer of stage Ⅲ-Ⅳ （t =-14.169,-15.509,-8.149,-8.509,P 〈 0.05）.The number of cells with positive expressions of IL-6,CD68 and D2-40 and positive co-expression of IL-6 and CD68 in the patients with infiltrative depth of stage T1-T2 were 41.5 ± 12.2,22.2 ±5.1,5.5 ± 2.6 and 9.7 ± 1.8,respectively,which were significantly different from 50.1 ± 8.8,27.3 ± 5.1,8.3 ± 3.9 and 12.0± 1.9 in the patients with infiltrative depth of stage T3-T4 （t =-2.435,-2.770,-2.212,-3.236,P 〈 0.05）.The number of cells with positive expressions of IL-6,CD68 and D2-40 and positive co-expression of IL-6 and CD68 in the patients without lymph node metastases were 39.2 ±6.7,21.3 ±3.0,5.2 ±3.3 and 9.8 ± 1.4,respectively,which were significantly different from 55.2 ± 6.4,29.9 ± 3.8,9.6 ± 2.9 and 12.7 ± 1.6 in the patients with lymph node metastases （t =-7.722,-7.838,-4.581,-5.756,P 〈 0.05）.All the 40 patients were followed up for 6-22 months with a median time of 18 months.The 1-year cumulative survival rate in the 20 patients with high expression of IL-6 was 70.0%,which was significantly different from 90.0% in the 20 patients with low expression of IL-6 （χ^2=4.410,P 〈 0.05）.The 1-year cumulative survival rate in the 20 patients with high expression of CD68 was 70.0%,which was significant different from 90.0% in the 20 patients with low expression of CD68 （χ^2 =4.075,P 〈 0.05）.The 1-year cumulative survival rate in the 20 patients with high expression of D2-40 was 65.0%,which was significantly different from 95.0% in the 20 patients with low expression of D2-40 （χ^2 =4.705,P 〈 0.05）.The 1-year cumulative survival rate in the 20 patients with high co-expression of IL-6 and CD68 was 70.0%,which was significantly different from 90.0% in the 20 patients with low co-expression of IL-6 and CD68 （χ^2=4.152,P 〈 0.05）.Conclusions There are high expressions of IL-6,TAMs and lymphangiogenesis in the tissues of gastric cancer,and positive correlations among expressions of IL-6,TAMs and lymphangiogenesis.The expressions of IL-6,TAMs and lymphangiogenesis in the tissues of gastric cancer are significantly correlated with the progression of gastric cancer and prognosis of paitents.