Ape1in-13(F13A)对M2型肿瘤相关巨噬细胞诱导的乳腺癌细胞侵袭的影响及机制

摘要

Objective To investigate effect of putative receptor protein related to the angiotensin receptor AT1 (APJ) antagonist Apelin13 (F13A) on the polarization of tumor-associated macrophages (TAM) and the invasion induced by M2 type macrophages in breast cancer cells line MCF-7 cells.Methods Macrophage RAW264.7 cells were treated with IL-13 (10 ng/mL) for 24 h to induce M2 type polarization (M2RAW) and the cells were treated with Apelin-13(F13A) (10.00 pm/mL) for 24 h.RAW and MCF-7 cells were co-cuhured for 48 h in non-contact manner.The proliferation of RAW and MCF-7 was detected by MTT.Levels of transforming growth factor-β (TGF-β) and interleukin-10 (IL-10) in culture medium were tested by ELISA.Expression of marker protein of M2 type polarization arginase 1 (Arg-1) were measured by Western blot.Migration and invasiveness of MCF-7 cells were tested by scratch test and Transwell method.Results Compared with RAW group,levels of TGF-β and IL-10 and expression of Arg-1 in M2RAW group were significantly increased,the differences were statistically significant (P ＜ 0.05).Compared with M2RAW group,levels of TGF-β and IL-10 and expression of Arg-1 in M2RAW+Apelin-13 (F13A) group were significantly decreased,the differences were statistically significant (P ＜ 0.05).Compared with RAW+MCF-7 group,scratch area was significantly decreased,the number of cell through the membrane was significantly increased in M2RAW+MCF-7 group,the differences were statistically significant (P ＜ 0.05).Compared with M2RAW+MCF-7 group,scratch area was significantly increased,the number of cell through the membrane was significantly decreased in Apelin-13(F13A)+M2RAW+ MCF-7 group,the differences were statistically significant (P ＜ 0.05).Conclusion Apelin13 (F13A) inhibits the polarization of tumor related macrophages and the invasion induced by M2 type macrophages in breast cancer cells line MCF-7 cells.%目的 探讨血管紧张素受体AT1相关的受体蛋白(APJ)拮抗剂Apelin13(F13A)对肿瘤相关巨噬细胞极化及其诱导的乳腺癌细胞系MCF-7细胞迁移和侵袭的影响.方法 用白细胞介素-13 (IL-13)(10 ng/mL)处理RAW264.7细胞24h诱导M2型极化(M2RAW),同时用Apelin-13 (F13A)(10.00 pm/mL)共同处理24h,再与MCF-7细胞非接触共培养48 h.采用MTT法检测RAW和MCF-7细胞的增殖.采用酶联免疫吸附测定法检测RAW细胞培养液上清中TGF-β和IL-10水平.Western blot检测RAW细胞中M2型极化标志蛋白Arg-1表达.划痕实验和Transwell实验检测MCF-7细胞的迁移和侵袭力.结果 与RAW组比较,M2RAW组TGF-β、IL-10及Arg-1的表达水平均显著增加,差异均有统计学意义(P＜ 0.05);与M2RAW组比较,M2RAW+Apelin-13(F13A)组TGF-β、IL-10水平及Arg-1的表达水平显著降低,差异均有统计学意义(P＜0.05).与RAW+MCF-7组比较,M2RAW+MCF-7组MCF-7细胞划痕显著减小,侵袭的细胞数明显增加,差异均有统计学意义(P＜0.05);与M2RAW+MCF-7组比较,Apelin-13(F13A)+M2RAW+MCF-7组MCF-7细胞划痕显著增加,侵袭的细胞数明显减少,差异均有统计学意义(P＜0.05).结论 Apelin13(F13A)可抑制巨噬细胞M2型极化及其诱导的乳腺癌细胞迁移和侵袭.