| Objective:To study the quantity of different phenotype of macrophages in gastric carcinomas and explore the relationship between it and the clinical pathologic stages of gastric carcinomos.Methods:Specimens of human gastric carcinomas and normal tissues6centimeter adjacent to carcinomas with different clinical stages(50cases of15in stage I,10in stage Ⅱ,14in stage Ⅲ,11in stageIV) were collected in Qingdao municipal hospital from March2012to December2012. All patients were divided into Ⅰ+Ⅱ stages group, Ⅲ+Ⅳ stages group, and normal tissues were set as control group.①Part of the gastric tissue was stained by immunohistochemical method, immunohistochemistry-SP was used to detected the quantity of CD68+and CD163+macrophages in gastric cancer and adjacent normal tissue.②Cell culture was employed to another part of the gastric tissue, digesting gastric cancer and adjacent normal tissue to obtain the cell suspension and use density gradient centrifugation to isolate macrophages. The macrophages were identified by morphological observation and immunofluorescence staining. Use Enzyme linked immunosorbent assay (ELISA) to detection the amount of the IL-10and IL-12secreted by macrophages in different time points. The relationship between the phenotypes of macrophages and the clinicopathologic features of gastric carcinomas was analyzed.Results:By Immunohistochemistry the number of CD68+macrophages in gasric tissue in control group, Ⅰ+Ⅱ and Ⅲ+Ⅳ stages group were4.72±1.40,14.98±2.19and18.54±2.54respectively. The difference was statistically significant between the three groups (x2=192.157,P<0.01),and the number of the macrophages was increased along with the higher clinical stage of gastric cancers (Z=-9.812,P<0.01).The number of CD163+macrophages in control group, Ⅰ+Ⅱ and Ⅲ+Ⅳstages group were3.16±1.18±5.21±2.95,12.42±2.78respectively. The number of macrophages in Ⅲ+Ⅳstages was larger than that in control and Ⅰ+Ⅱ stages group (Z=-10.396、12.317, P<0.01). CD68+/CD163macrophages in control group, Ⅰ+Ⅱ and Ⅲ+Ⅳstages group were9.66±1.42,5.90±1.50and1.72±0.68respectively. The difference between the three groups was significantly (F=131.253,P<0.01), and the number of the macrophages was decreased along with the higher clinical stage of gastric cancers(F=131.253,P<0.01). The number of CD68+and CD163+macrophages in the Lymph node metastasis group was higher than the group without lymph node metastasis (P<0.05). But the number of CD68+and CD163+macrophages had no relationship with the gender, age and tumor size. The number of CD68+and CD163+macrophages was respectively positive correlated with the clinical stage of gastric cancer (r=0.679, P<0.001),(r=0.917, P<0.001), and the number of CD68+/CD163macrophages was negative correlation with the clinical stage of gastric cancer (r=-0.938, P<0.001). Detected by ELISA in control and Ⅰ+Ⅱ stages group, the secretion quantity of IL-10all were below III+IV stages group (P<0.05), while the secretion quantity of IL-12all were higher than III+IV stages group (P<0.05), and the secretion of IL-10and IL-12showed a negative correlation (r=-0.512, P<0.001), The IL-12secreted by macrophages in gastric cancer was negative correlated with the clinical stages of gastric cancers (r=-0.671, P<0.001), and the secretion of IL-10was positive correlation with the clinical stages of gastric cancers (r=0.733, P<0.001).Conclusion:The macrophages of gastric cancers were mainly M1type in Ⅰ and Ⅱ clinical stages group as they secreted more IL-12; and the macrophages in gastric clinical stages Ⅲ and Ⅳ group were mainly M2type as they mainy secretion of IL-10, the macrophages were translated into M2phenotype with the development of gastric cancer,, within a condition that promote tumor growth. |
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