摘要
目的探讨肿瘤相关巨噬细胞(TAMs)上调miR-221-3p可能参与促进前列腺癌(PCa)恶性转移的作用机制。方法选择6例转移性PCa患者癌组织进行微小RNA(miRNAs)表达谱测序和差异表达miRNAs分析。分离上述转移性PCa患者癌组织中的原代TAMs。采用聚合酶链反应(qPCR)测定其中miR-221-3p的表达水平。向RAW264.7巨噬细胞转染miR-221-3p模拟物(mimic)或抑制物(inhibitor),然后与人PCa细胞系PC3细胞建立共培养体系。CCK-8法测定PC3细胞增殖能力,流式细胞术(FCM)测定细胞凋亡率,Transwell法测定细胞迁移和侵袭能力,Western blot法测定细胞上皮-间充质转化(EMT)相关蛋白因子的表达水平。结果在6例转移性PCa患者的癌组织中,与淋巴结转移阴性PCa组织来源TAMs相比,淋巴结转移阳性PCa组织来源TAMs中hsa-miR-221-3p显著上调(P<0.05)。在共培养体系中,与Mimic-NC组相比,miR-221-3p mimic组PC3细胞增殖能力明显增强;迁移能力和侵袭能力明显增强;EMT相关蛋白因子表达水平明显增强(除E-Cadherin)(P均<0.05)。与Inhibitor-NC组相比,miR-221-3p inhibitor组细胞增殖能力明显降低,凋亡率明显升高;迁移能力和侵袭能力明显被抑制;EMT相关蛋白因子表达水平明显被抑制(除E-Cadherin)(P均<0.05)。结论TAMs上调miR-221-3p可能参与促进了PCa恶性转移。
Objective To investigate the mechanism by which the up-regulation of miR-221-3p by tumor-associated macrophages(TAMs)may be involved in promoting the malignant metastasis of prostate cancer(PCa).Methods The microRNAs(miRNAs)expression profiles of 6 cases of metastatic PCa tissues were sequenced and analyzed.The primary TAMs were isolated.The expression of miR-221-3p was determined with qPCR.The miR-221-3p mimic or miR-221-3p inhibitor was transfected into RAW264.7 macrophages in vitro,and co-cultured with human prostate cancer PC3 cells.The proliferation,apoptosis,invasion and migration of PC3 cells were detected with CCK-8,flow cytometry(FCM),Transwell assay,respectively.Expressions of epithelial-mesenchymal transformation(EMT)related protein factors were determined with Western blot.Results In the 6 cases of metastatic PCa,hsa-miR-221-3p was significantly up-regulated in TAMs-derived from PCa tissues with positive lymph node metastasis(P<0.05).In the co-cultured system,compared with Mimic-NC group,miR-221-3p mimic group had significantly up-regulated proliferation,migration,invasion and EMT-related protein factors(except E-Cadherin)(P<0.05).Compared with Inhibitor-NC group,miR-221-3p inhibitor group had significantly up-regulated apoptosis rate,but down-regulated proliferation,migration,invasion and EMT-related protein factors(except E-Cadherin)(P<0.05).Conclusion The miR-221-3p expression up-regulate by TAMs may participate in the malignant metastasis of prostate cancer.
作者
阿斯木江·阿不拉
高新
宋光鲁
ASIMUJIANG Abula;GAO Xin;SONG Guanglu(Urology Center,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830000,China)
出处
《现代泌尿外科杂志》
CAS
2024年第4期368-374,378,共8页
Journal of Modern Urology
基金
新疆维吾尔自治区自然科学基金项目(No.2019D01C314)。
