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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。
The aim of this study was to investigate the effects of leptin on the migration and invasion of breast cancer cell line MCF7 induced by tumor-associated macrophages (TAMs) and its possible mechanism. The expression of CD206, TGF-β and IL-10 in THP1-differentiated macrophages were detected by RT-PCR, FQ-PCR and Western blot. The expression of Ob-Rb, a receptor for leptin, and Ob-Rt, a short receptor, were detected by RT-PCR in TAMs. Cell scratch assay and Transwell invasion assay were used to detect the migration and invasion ability of MCF cells. Western blot was used to detect the expression of p-STAT3, p-ERK 1/2 and p-AKT in TAMs. The expression of MMP2 and MMP9 in TAMs was detected by RT-PCR and Western blot. The results showed that the molecular phenotype of macrophages induced by 100 nmol / L PMA and 20 ng / mL IL-4 was CD206 + TGF-β HighIL-10 High. The leptin long receptor Ob-Rb and short receptor Ob-Rt in TAMs were highly expressed. Leptin-stimulated TAMs conditioned medium significantly enhanced the ability of MCF cells to migrate and invade. Leptin could significantly increase the expression of p-STAT3, p-ERK 1/2 and p-AKT in TAMs (P <0.05), and leptin upregulated the expression of MMP2 and MMP9 in TAMs; while MAPK / ERK 1/2 signaling PD98059 could inhibit the expression of MMP2, and AG490, an inhibitor of JAK / STAT signaling pathway, inhibited the expression of MMP9 (P <0.05). These results suggest that leptin can promote the migration and invasion of MCF7 cells through the activation of TAMs. The mechanism may be related to leptin upregulation of MMP2 in TAMs through MAPK / ERK 1/2 signaling pathway and up-regulation of MMP9 expression through JAK / STAT signaling pathway.