M2型巨噬细胞对食管癌移植瘤脉管生成的影响*
Effect of M2 type macrophage on angiogenesis and lymphangiogenesis of human esophageal cancer xenograft

目的:探讨M2型巨噬细胞对食管癌移植瘤脉管生成的影响及其可能机制。方法:应用PMA及IL-4将人淋巴瘤U937单核细胞诱导为M2型巨噬细胞。16只裸鼠分为2组,每组8只,注射与M2型巨噬细胞共培养的EC9706细胞或单独注射EC9706细胞; 观察2组荷瘤裸鼠成瘤情况,采用原位杂交法检测移植瘤中VEGF和 VEGF-C mRNA的表达,采用免疫组化方法检测2组移植瘤中VEGF和VEGF-C蛋白的表达,分别用CD31、D2-40标记2组移植瘤中血管和淋巴管,计算MVD与LMVD值。结果:M2型巨噬细胞与EC9706细胞共培养组裸鼠移植瘤中VEGF、VEGF-C mRNA及蛋白表达水平高于单独注射EC9706细胞的对照组(P<0.05), MVD及LMVD值亦高于对照组(P<0.05)。结论:M2型巨噬细胞可通过分泌VEGF、VEGF-C促进裸鼠食管癌移植瘤中血管、淋巴管的生成。
Aim: To investigate the effect of M2 type macrophage on angiogenesis and lymphangiogenesis of human esophageal cancer xenograft.Methods: Human lymphoma U937 monocytes were induced into M2 type macrophage by PMA and IL-4. The M2 type macrophage was co-cultured with esophageal cancer EC9706 cells. Sixteen nude mice were allocated into two groups and were injected with co-culture M2 type macrophage with EC9706 cells(co-culture group)or single-culture EC9706 cells(control group). The formation of tumor in the 2 groups was observed. The mRNA expression levels of VEGF and VEGF-C in xenograft of the 2 groups were determined by in-situ hybridization method. The levels of VEGF and VEGF-C proteins in xenograft of the 2 groups were detected by immunohistochemical technique. The expression levels of CD31 and D2-40 separately to mark the blood vessel and the lymphatic vessel were detected by immunohistochemistry, and the value of MVD and LMVD was calculated.Results: The protein and mRNA expression levels of VEGF and VEGF-C in co-culture group were significantly higher than those in control group(P<0.05). Both the value of MVD and that of LMVD in co-culture group were also higher than those in control group(P<0.05).Conclusion: M2 type macrophage may promote angiogenesis and lymphatic vessel generation of esophageal cancer xenograft in nude mice by secreting VEGF and VEGF-C