乳腺癌细胞LncRNA OIP5-AS1对巨噬细胞极化的影响以及极化巨噬细胞对乳腺癌细胞的反馈调节OA
Effect of LncRNA OIP5-AS1 in Breast Cancer Cells on Macrophage Polarization and Feedback Regulation of Polarized Macrophages on Breast Cancer Cells
目的 探究乳腺癌源性Lnc-RNA 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Cont-rol组(空白对照)、NC 组(转染NC siRNA)和si-OIP5组(转染LncRNA OIP5-AS1 siRNA).RT-qPCR检测LncRNA OIP5-AS1、IL-4和IL-13 mRNA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RPMI 1640培养基中诱导M0巨噬细胞向TAM极化,RT-qPCR检测TAM中CD206 mRNA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MDA-MB-231细胞迁移侵袭能力并检测Vimentin、N-cadherin、E-cadherin表达变化.结果 相较于Control及NC组,si-OIP5组LncRNA OIP5-AS1表达显著下调(P<0.001),IL-13 mRNA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Vim-entin表达下调(均P<0.01).结论 乳腺癌源性LncRNA 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.
Objective To explore the mechanism by which breast cancer-derived LncRNA 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 siRNA),and the si-OIP5 group(transfected with LncRNA OIP5-AS1 siRNA).The mRNA expression levels of LncRNAs OIP5-AS1,IL-4,and IL-13 were detected by RT-qPCR.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 mRNA expression of CD206 in TAMs was detected by RT-qPCR.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,N-cadherin and E-cadherin.Results Compared with the control and NC groups,the si-OIP5 group showed significantly downregulated expression of LncRNA OIP5-AS1(P<0.001)and significantly decreased IL-13 mRNA 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 volume-dependent 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 LncRNA OIP5-AS1 can induce TAM M2 polarization mediated by IL-13,thereby promoting the migration,invasion,and EMT of breast cancer cells.
杨恩帅;董哲;常鑫悦;肖子阳;刘洋;郭素芬
157011 牡丹江,牡丹江医科大学病理教研室||157011 牡丹江,黑龙江省肿瘤疾病防治重点实验室157011 牡丹江,牡丹江医科大学病理教研室||157011 牡丹江,黑龙江省肿瘤疾病防治重点实验室157011 牡丹江,牡丹江医科大学病理教研室||157011 牡丹江,黑龙江省肿瘤疾病防治重点实验室157011 牡丹江,牡丹江医科大学病理教研室||157011 牡丹江,黑龙江省肿瘤疾病防治重点实验室157011 牡丹江,牡丹江医科大学病理教研室||157011 牡丹江,黑龙江省肿瘤疾病防治重点实验室157011 牡丹江,牡丹江医科大学病理教研室||157011 牡丹江,黑龙江省肿瘤疾病防治重点实验室
医药卫生
乳腺癌M2型巨噬细胞LncRNA OIP5-AS1迁移上皮间质转化
Breast neoplasmsM2 macrophagesLncRNA OIP5-AS1MigrationEpithelium-mesenchymal transformation
《肿瘤防治研究》 2026 (3)
187-193,7
Doctoral Research Startup Fund of Mudanjiang Medical University(No.2021-MYBSKY-012)Natural Science Foundation of Heilongjiang Province of China(No.LH2019H199) 牡丹江医科大学博士启动基金(2021-MYBSKY-012)黑龙江省自然科学基金(LH2019H199)
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