乳腺癌细胞外泌体miR-27a对巨噬细胞极化及肿瘤生长和转移的影响

Effects of breast cancer cell exosomes miR-27a on the polarization of macrophages and on tumor growth and metastasis

目的 探究乳腺癌细胞外泌体miR-27a对巨噬细胞极化及肿瘤生长和转移的影响。方法 实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测M1型巨噬细胞和M2型巨噬细胞中miR-27a的表达,并且检测miR-27a对巨噬细胞极化的影响,差速离心法提取乳腺上皮细胞MCF10A和乳腺癌细胞MDA-MB-231外泌体,qRT-PCR检测外泌体中miR-27a的表达,检测外泌体miR-27a对巨噬细胞极化的影响,取外泌体诱导后的巨噬细胞培养基上清与MDA-MB-231细胞共孵育后,检测MDA-MB-231细胞增殖、迁移和侵袭能力。结果 miR-27a高表达于M2型巨噬细胞(P<0.05),并且miR-27a mimic显著促进CD206和MRC-2表达(P<0.05),miR-27a inhibitor抑制CD206和MRC-2表达(P<0.05),miR-27a在MDA-MB-231外泌体中显著高表达(P<0.05),MDA-MB-231外泌体可促进M2型巨噬细胞极化(P<0.05),极化后的巨噬细胞培养上清可显著诱导MDA-MB-231细胞的增殖、迁移和侵袭能力增加(P<0.05),miR-27a低表达的MDA-MB-231外泌体可显著抑制M2型巨噬细胞极化(P<0.05),并且该M2型细胞培养上清可显著抑制MDA-MB-231细胞的增殖、迁移和侵袭能力(P<0.05)。结论 乳腺癌细胞外泌体miR-27a可促进M2型巨噬细胞极化而促进乳腺癌细胞的生长和转移。

Objective To explore the effects of breast cancer cell exosomes miR-27 a on the polarization of macrophages and on tumor growth and metastasis.Methods qRT-PCR was used to detect the expression of miR-27 a in M1 and M2 macrophages, and to detect the effect of miR-27 a on the polarization of macrophages.Differential centrifugation was used to extract breast epithelial cells MCF10 A and breast cancer cells MDA-MB-231 exosomes, qRT-PCR to detect the expression of miR-27 a in exosomes, to detect the effect of exosomes miR-27 a on the polarization of macrophages, and culture the macrophages after exosomes induction After the base supernatant(CM) was incubated with MDA-MB-231 cells, the proliferation, migration and invasion ability of MDA-MB-231 cells were detected.Results miR-27 a was highly expressed in M2 type macrophages(P<0.05),and miR-27 a mimic significantly promoted the expression of CD206 and MRC-2(P<0.05),and miR-27 a inhibitor inhibited the expression of CD206 and MRC-2(P<0.05),miR-27 a was highly expressed in MDA-MB-231 exosomes(P<0.05),MDA-MB-231 exosomes can significantly promoted the polarization of M2 macrophages(P<0.05).The polarized macrophage culture supernatant could significantly induce the proliferation, migration and invasion of MDA-MB-231 cells(P<0.05),and MDA-MB-231 exosomes with low expression of miR-27 a could significantly inhibit M2 Type macrophages were polarized(P<0.05),and the M2 type cell culture supernatant could significantly inhibit the proliferation, migration and invasion of MDA-MB-231 cells(P<0.05).Conclusion The breast cancer cell exosomes miR-27 a can promote the polarization of M2 macrophages and promote the growth and metastasis of breast cancer cells.