Department of Clinical Laboratory, Cancer Hospital, Harbin Medical University, Harbin, China.
Department of Clinical Laboratory, Nanjing Maternity and Child Health Care Hospital, Nanjing Medical University, Nanjing, China.
Cancer Gene Ther. 2015 Nov;22(11):518-23. doi: 10.1038/cgt.2015.45. Epub 2015 Oct 23.
Bone marrow-derived endothelial progenitor cells (EPCs) migrate to and engraft at ovarian cancer sites. Understanding the interactions between ovarian cancer cells and EPCs is fundamental for determining whether to harness EPC-tumor interactions for delivery of therapeutic agents or target them for intervention. Ovarian cancer cell lines (SKOV-3 and OVCAR-3) were cultured alone or in EPC-conditioned media (EPC-CM). Migration of ovarian cancer cells was detected by transwell chamber. N-cadherin and E-cadherin expression were analyzed by real-time reverse transcription PCR and western blot. EPC-CM can increase transforming growth factor-beta (TGF-β) secretion in SKOV-3 and OVCAR-3 cells. EPC-CM induced loss of ovarian cancer cell-cell junctions, downregulation of E-cadherin, upregulation of N-cadherin and acquisition of a fibroblastic phenotype, consistent with an epithelial-to-mesenchymal transition (EMT). The specific TGF-β inhibitor SB431542 abolished the SKOV-3 and OVCAR-3 ovarian cancer cell migration induced by EPC-CM. In SKOV-3 and OVCAR-3 cells, EPC-CM downregulated E-cadherin and concurrently upregulated N-cadherin. EPC-CM upregulated the expression of transcriptional repressors of E-cadherin, Snail and Twist. Treatment with SB431542 abolished the effects of EPC-CM on the relative expression levels of cadherin, Snail and Twist. This study demonstrates that TGF-β has a role in EPC-CM-induced ovarian cancer migration by activating EMT.
骨髓来源的内皮祖细胞(EPC)迁移到卵巢癌细胞并在其部位定植。了解卵巢癌细胞与 EPC 之间的相互作用对于确定是否利用 EPC-肿瘤相互作用来输送治疗剂或针对它们进行干预至关重要。单独培养卵巢癌细胞系(SKOV-3 和 OVCAR-3)或在 EPC 条件培养基(EPC-CM)中培养。通过 Transwell 室检测卵巢癌细胞的迁移。通过实时逆转录 PCR 和 Western blot 分析 N-钙粘蛋白和 E-钙粘蛋白的表达。EPC-CM 可增加 SKOV-3 和 OVCAR-3 细胞中转化生长因子-β(TGF-β)的分泌。EPC-CM 诱导卵巢癌细胞细胞-细胞连接丢失,E-钙粘蛋白下调,N-钙粘蛋白上调,并获得成纤维细胞表型,与上皮-间充质转化(EMT)一致。特异性 TGF-β抑制剂 SB431542 可消除 EPC-CM 诱导的 SKOV-3 和 OVCAR-3 卵巢癌细胞迁移。在 SKOV-3 和 OVCAR-3 细胞中,EPC-CM 下调 E-钙粘蛋白,同时上调 N-钙粘蛋白。EPC-CM 上调 E-钙粘蛋白转录抑制因子 Snail 和 Twist 的表达。用 SB431542 处理可消除 EPC-CM 对钙粘蛋白、Snail 和 Twist 相对表达水平的影响。本研究表明,TGF-β 通过激活 EMT 在 EPC-CM 诱导的卵巢癌细胞迁移中起作用。
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