27-Hydroxycholesterol-induced EndMT acts via STAT3 signaling to promote breast cancer cell migration by altering the tumor microenvironment
27-Hydroxycholesterol-induced EndMT acts via STAT3 signaling to promote breast cancer cell migration by altering the tumor microenvironment
复制标题
27-羟基胆固醇诱导的 EndMT 通过 STAT3 信号传导发挥作用，通过改变肿瘤微环境来促进乳腺癌细胞迁移
DOI：
10.20892/j.issn.2095-3941.2019.0262
复制
发表时间：
2020-02-01
影响因子：
5.5
通讯作者：
Li, Zhong
中科院分区：
文献类型：
--
作者：
Jiao, Kailin;Zhen, Jing;Li, Zhong
目的：内皮间质转化（EndMT）通过调节肿瘤微环境（TME）的复杂性在癌症转移中发挥重要作用。在这里，我们研究了 27-羟基胆固醇 (27HC) 是否诱导内皮细胞 (EC) 中的 EndMT。方法：用蛋白质印迹法评估人微血管内皮细胞 1 (HMEC-1) 细胞系和经 27HC 刺激的人脐静脉内皮细胞 (HUVEC) 中的 EndMT 标记。采用定量实时聚合酶链反应 (qRT-PCR) 研究条件培养基中培养的乳腺癌 (BC) 细胞的上皮间质转化 (EMT) 标记。分别采用 qRT-PCR 和明胶酶谱法检测 MMP-2 和 MMP-9 mRNA 表达和活性。通过蛋白质印迹、免疫荧光染色和细胞转染测定验证了激活的 STAT3 对 2711C 诱导的 EndMT 的影响。通过Transwell实验评估BC细胞的迁移能力。结果：我们发现27HC诱导HMEC 1和HUVEC中的EndMT，并且27HC诱导的EndMT促进EMT和BC细胞迁移。 27HC诱导的BC细胞EMT也促进EndMT和HUVEC迁移。对潜在分子机制的研究表明，STAT3 敲除抑制了 HUVEC 中的 EndMT 以及 27HC 诱导的 BC 细胞中的迁移。此外，C646 和白藜芦醇（STAT3 乙酰化抑制剂）可抑制暴露于 27HC 的 HUVEC 中 Ac-STAT3、p-STAT3 和 EndMT 标记物的表达。这些HUVEC反过来减弱了27HC诱导的EndMT中BC细胞的迁移能力。结论：在TME中观察到27HC诱导的EndMT和EMT之间的串扰。此外，发现 STAT3 信号传导的激活参与 27HC 诱导的 EndMT。
Objective: The endothelial to mesenchymal transition (EndMT) plays a major role in cancer metastasis by regulating the complexity of the tumor microenvironment (TME). Here, we investigated whether 27-hydroxycholesterol (27HC) induces EndMT in endothelial cells (ECs).Methods: EndMT markers in the human microvascular endothelial cell-1 (HMEC- 1) cell line and human umbilical vein endothelial cells (HUVECs) stimulated with 27HC were evaluated with Western blot. Epithelial to mesenchymal transition (EMT) markers in breast cancer (BC) cells cultured in conditioned medium were investigated with quantitative real time polymerase chain reaction (qRT-PCR). The MMP-2 and MMP-9 mRNA expression and activity were detected with qRT-PCR and gelatin zymography assays, respectively. The effect of activated STAT3 on 2711C-induced EndMT was validated by Western blot, immunofluorescence staining, and cell transfection assays. The migration ability of BC cells was evaluated with Transwell assays.Results: We found that 27HC induced EndMT in HMEC 1 and HUVECs, and 27HC induced EndMT facilitated EMT and BC cell migration. The 27HC-induced EMT of BC cells also promoted EndMT and HUVEC migration. Investigation of the underlying molecular mechanisms revealed that STAT3 knockdown repressed EndMT in HUVECs as well as migration in BC cells induced with 27HC. In addition, C646 and resveratrol, inhibitors of STAT3 acetylation, repressed the expression of Ac-STAT3, p-STAT3, and EndMT markers in HUVECs exposed to 27HC; these HUVECs in turn attenuated the migration ability of BC cells in 27HC-induced EndMT.Conclusions: Cross-talk between 27HC-induced EndMT and EMT was observed in the TME. Moreover, activation of STAT3 signaling was found to be involved in 27HC-induced EndMT.