瘦素激活肿瘤相关巨噬细胞促进乳腺癌细胞MCF7的迁移及侵袭

Tumor-associated Macrophages Activated by Leptin Promote Migration and Invasion of Breast Cancer MCF7 Cells

该文探讨瘦素(leptin)激活肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)对乳腺癌细胞MCF7迁移及侵袭的影响及其作用机制。RT-PCR、FQ-PCR及Western blot检测THP1分化的巨噬细胞中CD206、TGF-β及IL-10的表达。RT-PCR检测TAMs中leptin长受体Ob-Rb及短受体Ob-Rt的表达。细胞划痕试验和Transwell侵袭试验检测MCF细胞的迁移及侵袭能力。Western blot检测TAMs中p-STAT3、p-ERK 1/2和p-AKT的表达。RT-PCR及Western blot检测TAMs中MMP2、MMP9的表达。结果表明,经100 nmol/L PMA及20 ng/mL IL-4诱导成的巨噬细胞分子表型为CD206+TGF-βHighIL-10High。TAMs中leptin长受体Ob-Rb及短受体Ob-Rt均为高表达。经leptin刺激的TAMs条件培养基能明显增强MCF细胞的迁移及侵袭能力。Leptin能显著提高TAMs中p-STAT3、p-ERK 1/2和p-AKT的表达(P<0.05),且leptin能上调TAMs中MMP2和MMP9的表达;而MAPK/ERK 1/2信号通路抑制剂PD98059能抑制MMP2的表达,JAK/STAT信号通路抑制剂AG490能抑制MMP9的表达(P<0.05)。以上结果表明,leptin能通过激活TAMs促进MCF7细胞的迁移和侵袭,其机制可能与leptin通过MAPK/ERK 1/2信号通路上调TAMs中MMP2及通过JAK/STAT信号通路上调MMP9的表达有关。

This paper investigated the effect of tumor-associated macrophages activated by leptin on migra- tion and invasion of MCF7 cells and explored its molecule mechanisms. The expressions of CD206, TGF-β1and IL- l0 were detected by RT-PCR, FQ-PCR and Western blot. Ob-Rb and Ob-Rt in TAMs were detected by RT-PCR. The migration and invasion of MCF7 cells were determined by cell scratch assay and Transwell chamber assay. The expressions of p-STAT3, p-ERK 1/2 and p-AKT in TAMs were detected by Western blot. The mRNA and protein expression of MMP2 and MMP9 in TAMs were detected by RT-PCR and Western blot. The results suggested that the phenotype of macrophage induced by PMA （100 nmol/L） and IL-4 （20 ng/mL） was CD206^＋TGF-β^HighIL-10^High. The conditional medium of TAMs activated by leptin significantly increased the migration and invasion of MCF7 cells. The expressions ofp-STAT3, p-ERK 1/2 and p-AKT in TAMs were significantly enhanced by leptin （P〈0.05）. Furthermore, the mRNA and protein expressions of MMP2 and MMP9 in TAMs were remarkably up-regulated by leptin （P〈0.05）. However, the MAPK/ERK 1/2 inhibitor PD98059 could down-regulate the expression of MMP2 in TAMs treated with leptin （P〈0.05）, and JAK/STAT inhibitor AG490 could decrease the expression of MMP9 （P〈0.05）. In conclusion, leptin can enhance the migration and invasion of MCF7 cells via activating TAMs, which may be associated with the up-regulation of MMP2 and MMP9 in TAMs, through MAPK/ERK 1/2 and JAK/STAT signaling pathways, respectively.