胃癌外泌体lncRNA MM2P诱导STAT6磷酸化而促进M2型巨噬细胞极化

Gastric cancer exosomal lncRNA MM2P induces phosphorylation of STAT6 and promotes polarization of M2 type macrophages

目的探究胃癌外泌体lncRNA MM2P对巨噬细胞极化的影响及机制。方法诱导THP1细胞分化为M0、M1、M2型巨噬细胞,qRT-PCR检测各组巨噬细胞中lncRNA MM2P表达水平,提取并鉴定胃黏膜上皮细胞GES-1和胃癌细胞株MGC-803、SGC-7901所分泌的外泌体(GES-1 exo、MGC-803 exo、SGC-7901 exo),qRT-PCR检测lncRNA MM2P在胃癌细胞及其外泌体中的表达,将lncRNA MM2P及Vector转染至M0型巨噬细胞,qRT-PCR检测细胞中M2型巨噬细胞标志蛋白CD206、CD163、IL-10的表达,将胃癌细胞外泌体(MGC-803 exo和SGC-7901 exo)及差异表达lncRNA MM2P的外泌体(MGC-803/Vector exo和MGC-803/lncRNA MM2P exo)与M0型巨噬细胞共孵育后,qRT-PCR检测CD206、CD163、IL-10的表达,Western blotting检测STAT1和STAT6磷酸化水平。结果lncRNA MM2P高表达于M2型巨噬细胞(P<0.001),并且lncRNA MM2P在胃癌细胞株及其外泌体中的表达水平均显著高于其在胃黏膜上皮细胞及其外泌体中的表达(P<0.001)。与Vector组比较,转染lncRNA MM2P后,M2型巨噬细胞标志蛋白CD206、CD163、IL-10表达显著上调(P<0.001)。与GES-1 exo组比较,MGC-803 exo组和SGC-7901 exo组细胞CD206、CD163、IL-10表达显著上调(P<0.001),且MGC-803/lncRNA MM2P exo组细胞CD206、CD163、IL-10表达水平及STAT6磷酸化水平均显著高于MGC-803/Vector exo组。结论胃癌外泌体中lncRNA MM2P可显著促进M2型巨噬细胞极化,并且可促进STAT6磷酸化。

Objective To study the effect and mechanism of gastric cancer exosomal lncRNA MM2 P on the polarization of macrophages.Methods THP1 cells were induced to differentiate into M0,M1,M2 type macrophages,the expression levels of lncRNA MM2 P in each group of macrophages were detected by qRT-PCR.Exosomes secreted by gastric mucosal epithelial cells GES-1 and gastric cancer cell lines MGC-803,SGC-7901 were extracted and identified(GES-1 exo,MGC-803 exo,SGC-7901 exo),qRT-PCR was used to detect the expression of lncRNA MM2 P in gastric cancer cells and their exosomes,and transfected lncRNA MM2 P and Vector to M0 type macrophages,qRT-PCR was used to detect the expression of M2 type macrophage marker proteins CD206,CD163,and IL-10 in the cells.Gastric cancer cell exosomes(MGC-803 exo and SGC-7901 exo)and exosomes differentially expressed lncRNA MM2 P(MGC-803/Vector exo and MGC-803/lncRNA MM2 P exo)were incubated with M0 type macrophages,the expressions of CD206,CD163,IL-10 were detected by qRT-PCR,STAT1 and STAT6 phosphorylation level were detected by Western blotting.Results lncRNA MM2 P was highly expressed in M2 type macrophages(P<0.001),and the expression level of lncRNA MM2 P in gastric cancer cell lines and their exosomes was significantly higher than that in gastric mucosal epithelial cells and their exosomes(P<0.001).Compared with the Vector group,after transfection of lncRNA MM2 P,the expression of M2 type macrophage marker proteins CD206,CD163 and IL-10 was significantly up-regulated(P<0.001).Compared with the GES-1 exo group,the expressions of CD206,CD163,and IL-10 in the MGC-803 exo group and SGC-7901 exo group were significantly up-regulated(P<0.001),and the expression level of CD206,CD163,IL-10 and STAT6 phosphorylation in the MGC-803/lncRNA MM2 P exo group were significantly higher than those in MGC-803/Vector exo group.Conclusion The lncRNA MM2 P in gastric cancer exosomes can significantly promote the polarization of M2 type macrophages and promote the phosphorylation of STAT6.