M2型巨噬细胞通过分泌白细胞介素-10调控胆囊癌细胞增殖、迁移和侵袭的研究

M2 macrophages regulate the proliferation,migration and invasion of gallbladder cancer cells by secreting interleukin-10

目的:探讨M2型巨噬细胞通过调控白细胞介素-10(IL-10)表达影响胆囊癌(gallbladder cancer,GBC)细胞增殖、迁移、侵袭的研究。方法:将人单核细胞株THP-1在体外诱导成M2型巨噬细胞,利用RTqPCR检测相应标记物和IL-10的表达情况;利用ELISA检测细胞上清液中IL-10的表达情况。将GBC细胞株NOZ和GBC-SD分组为Control组(无血清培养基)、M2-CM组(M2型巨噬细胞条件培养基)、M2-CM+anti-IL-10组(含IL-10中和抗体的M2-CM)和M2-CM+IgG组(含IL-10同型对照抗体的M2-CM)。采用CCK-8实验、细胞划痕实验和Transwell实验检测各组细胞的增殖、迁移和侵袭变化状况,再用Western blot实验检测各组细胞的上皮间质转化标志物(E-cadherin和Vimentin)的表达量。结果:M2型巨噬细胞在体外诱导成功;与诱导前的THP-1相比,IL-10在M2型巨噬细胞内和上清液中表达明显升高(P<0.05);M2型巨噬细胞可促进GBC细胞增殖、迁移和侵袭,以及能抑制E-cadherin表达和促进Vimentin表达,阻断IL-10可抑制M2型巨噬细胞的这一影响(P<0.05)。结论:M2型巨噬细胞可通过分泌IL-10促进GBC细胞增殖、迁移、侵袭及上皮间质转化。

Objective:To investigate the effect of M2 macrophages on the proliferation,migration and invasion of gallbladder carcinoma cells by regulating the expression of interleukin-10(IL-10).Methods:Human monocyte line THP-1 was induced into M2 macrophages in vitro,and the expression of corresponding markers and IL-10 was detected by RT-qPCR.The expression of IL-10 in cell supernatant was detected by ELISA.GBC cell lines NOZ and GBC-SD were divided into the Control group(serumfree medium)、the M2-CM group(M2 macrophage conditioned medium)、the M2-CM+anti-IL-10 group(M2-CM containing anti-IL-10)and the M2-CM+IgG group(M2-CM containing IL-10 isotype control antibody).The cell proliferation rate of each group was measured by CCK-8 test,the cell migration rate of each group was detected by cell scratch test and the change in cell invasive ability of each group was detected by Transwell test;and Western blot assay was used to detect the expression of epithelialmesenchymal transformation markers(E-cadherin and Vimentin)in each group.Results:M2 macrophages were successfully induced in vitro.Compared with uninduced THP-1,IL-10 was highly expressed in M2 macrophages and M2 macrophages supernatant(P<0.05).M2 macrophages could promote the proliferation,migration and invasion of GBC cells,as well as inhibit the expression of E-cadherin and promote the expression of Vimentin.Blocking IL-10 could inhibit the effect of M2 macrophages(P<0.05).Conclusion:M2 macrophages can promote the proliferation,migration,invasion and epithelial-mesenchymal transformation of GBC cells by secreting IL-10.