【摘要】 目的 探究乳腺癌源性LncRNA OIP5-AS1经肿瘤相关巨噬细胞（TAM）M2型极化调控乳腺癌细胞迁移、侵袭及上皮—间质转化的机制。方法 MDA-MB-231细胞分设Control组（空白对照）、NC组（转染NC si RNA）和si-OIP5组（转染Lnc RNAOIP5-AS1 si RNA）。RT-qPCR检测Lnc RNA OIP5-AS1、IL-4和IL-13 m RNA表达；ELISA检测培养基上清IL-4和IL-13蛋白表达。Control组培养上清按不同体积比加入RPMI1640培养基中诱导M0巨噬细胞向TAM极化，RT-qPCR检测TAM中CD206 m RNA的表达；以NC组/si-OIP5组培养基上清诱导M0型巨噬细胞，Western blot检测CD206表达；继而构建巨噬细胞MDA-MB-231共培养模型，评估MDAMB-231细胞迁移侵袭能力并检测Vimentin、N-cadherin、Ecadherin表达变化。结果 相较于Control及NC组，si-OIP5组Lnc RNAOIP5-AS1表达显著下调（P<0.001）, IL-13m RNA及蛋白水平均显著降低（P<0.05），而IL-4表达在各组间无明显差异。条件培养基呈体积依赖性诱导M0巨噬细胞CD206表达，40%体积比时效应最强（P<0.001）。相较NC组，si-OIP5组CD206蛋白水平显著降低（P<0.01）。共培养体系中，si-OIP5组MDA-MB-231细胞迁移侵袭能力显著减弱（P<0.001）,E-cadherin表达上调，N-cadherin及Vimentin表达下调（均P<0.01）。结论 乳腺癌源性Lnc RNA OIP5-AS1可经IL-13介导诱导TAM M2型极化，进而促进乳腺癌细胞的迁移、侵袭及EMT。更多还原

【Abstract】 Objective To explore the mechanism by which breast cancer-derived Lnc RNA OIP5-AS1 regulates the migration, invasion, and epithelial-mesenchymal transition of breast cancer cells through the M2 polarization of tumor-associated macrophages(TAM). Methods MDA-MB-231 cells were divided into the control group(blank control), the NC group(transfected with NC si RNA), and the si-OIP5 group(transfected with Lnc RNA OIP5-AS1 si RNA). The m RNA expression levels of Lnc RNAs OIP5-AS1, IL-4, and IL-13 were detected by RT-q PCR. The protein expression levels of IL-4 and IL-13 in the culture supernatant were detected by ELISA. The culture supernatant from the control group was added to RPMI 1 640 medium at different volume ratios to induce the polarization of M0 macrophages into TAMs. The m RNA expression of CD206 in TAMs was detected by RT-q PCR. M0-type macrophages were induced with the medium supernatant of the NC group/si-OIP5 group, and the expression of CD206 was detected by Western blot.Subsequently, a co-culture model of macrophage and MDA-MB-231 was constructed to evaluate the migration and invasion abilities of MDA-MB-231 cells and detect the expression changes in Vimentin, Ncadherin and E-cadherin. Results Compared with the control and NC groups, the si-OIP5 group showed significantly downregulated expression of Lnc RNA OIP5-AS1(P<0.001) and significantly decreased IL-13m RNA and protein levels(P<0.05). Meanwhile, no significant difference was observed in the expression of IL-4 among the groups. Conditioned medium induced CD206 expression in M0 macrophages in a volumedependent manner, with the strongest effect at a 40% volume ratio(P<0.001). The CD206 protein level in the si-OIP5 group significantly decreased(P<0.01). In the co-culture system, the migration and invasion abilities of MDA-MB-231 cells in the si-OIP5 group significantly weakened(P<0.001), the expression of E-cadherin was upregulated, and the expression levels of N-cadherin and Vimentin were downregulated(both P<0.01).Conclusion Breast cancer-derived Lnc RNA OIP5-AS1 can induce TAM M2 polarization mediated by IL-13, thereby promoting the migration, invasion, and EMT of breast cancer cells.更多还原