lncRNA DLEU2通过调控巨噬细胞M2极化对卵巢癌细胞转移的影响

Effect of lncRNA DLEU2 on metastasis of ovarian cancer cells by regulating M2 polarization of macrophages

目的 探讨长链非编码RNA(lncRNA)DLEU2对卵巢癌细胞转移的影响。方法 采用实时荧光定量PCR(RT-qPCR)实验检测lncRNA DLEU2在卵巢癌组织和卵巢癌细胞SKOV3中的表达,Transwell实验和Western blot分别检测lncRNA DLEU2对SKOV3细胞迁移、侵袭能力以及上皮-间充质转化(EMT)相关蛋白N-cadherin、Vimentin、E-cadherin表达的影响。用佛波醇肉豆蔻酸乙酸酯(PMA)处理THP-1单核细胞诱导其分化为巨噬细胞。随后使用DLEU2-siRNA、NC-siRNA转染组SKOV3细胞的上清液处理巨噬细胞。收集处理巨噬细胞的条件培养基(CM),用其处理SKOV3细胞。将细胞分为Control组、NC-siRNA+CM组、DLEU2-siRNA+CM组。采用RT-qPCR检测iNOS、CD86、CD163、Arg-1和IL-10mRNA的水平。结果 与癌旁组织和人正常卵巢上皮细胞IOSE80比较,卵巢癌组织和SKOV3细胞中lncRNA DLEU2表达均显著升高(P<0.05)。敲低lncRNA DLEU2可显著抑制SKOV3细胞侵袭、迁移能力以及EMT相关蛋白的表达。M2型巨噬细胞中lncRNADLEU2的表达高于M1型巨噬细胞(P<0.05)。与Control组比较,NC-siRNA+CM组SKOV3细胞侵袭、迁移能力以及EMT相关蛋白的表达显著升高(P<0.05);与NC-siRNA+CM组比较,DLEU2-siRNA+CM组SKOV3细胞侵袭、迁移能力以及EMT相关蛋白的表达显著降低(P<0.05)。结论 敲低lncRNA DLEU2可通过调控M2型巨噬细胞极化抑制卵巢癌细胞转移。

Objective To investigate the effect of long non-coding RNA(lncRNA)DLEU2 on the metastasis of ovar-ian cancer cells.Methods Real-time quantitative PCR(RT-qPCR)was used to detect the expression of lncRNA DLEU2 in ovarian cancer tissues and SKOV3 cells.Transwell assay and Western blot were used to detect the effects of lncRNA DLEU2 on the migration and invasion ability of SKOV3 cells and the expression of epithelial-mesenchymal transition(EMT)-related proteins N-cadherin,Vimentin and E-cadherin.THP-1 monocytes were treated with phorbol myristate acetate(PMA)to induce differentiation into macrophages.Subsequently,the macrophages were treated with the supernatant of SKOV3 cells transfected with DLEU2-siRNA and NC-siRNA.The conditioned medium(CM)of macrophages was collected and used to treat SKOV3 cells.The cells were divided into Control group,NC siRNA+CM group and DLEU2 siRNA+CM group.RT-qPCR was used to detect the mRNA levels of iNOS,CD86,CD163,Arg-1 and IL-10.Results Compared with the adjacent tissues and human normal ovarian epithelial cells IOSE80,the expression of lncRNA DLEU2 was significantly increased in ovarian cancer tis-sues and SKOV3 cells(P<0.05).Knockdown of lncRNA DLEU2 significantly inhibited the invasion and migration of SKOV3 cells and the expression of EMT-related proteins.The expression of lncRNA DLEU2 in M2 type macrophages was higher than that in M1 type macrophages(P<0.05).Knockdown of lncRNA DLEU2 significantly reduced the expression of M2 markers CD163,Arg-1 and IL-10 mRNA in macrophages.Compared with the Control group,the invasion and migration ability of SKOV3 cells and the expression of EMT-related proteins were significantly increased in the NC-siRNA+CM group(P<0.05).Compared with NC siRNA+CM group,the invasion and migration ability of SKOV3 cells and the expression of EMT-related proteins were significantly decreased in DLEU2 siRNA+CM group(P<0.05).Conclusion Knockdown of lncRNA DLEU2 could inhibit the metastasis of ovarian cancer cells by regulating the polarization of M2 macrophages.