MicroRNA-126-5p downregulates BCAR3 expression to promote cell migration and invasion in endometriosis
MicroRNA-126-5p downregulates BCAR3 expression to promote cell migration and invasion in endometriosis
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MicroRNA-126-5p下调BCAR3表达促进子宫内膜异位症细胞迁移和侵袭
DOI：
10.1016/j.mce.2019.110486
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发表时间：
2019-08-20
影响因子：
4.1
通讯作者：
Wang, Danbo
中科院分区：
文献类型：
--
作者：
Meng, Xiannan;Liu, Jing;Wang, Danbo
目的：子宫内膜异位症（EMs）是一种雌激素依赖性多因素疾病。子宫内膜细胞中雌激素的抑制导致它们无法在异位部位形成病变。然而，减少或抑制雌激素的抑制作用是否会导致异位病变的建立仍不清楚。 BCAR3基因诱导雌激素依赖性乳腺癌细胞产生雌激素抵抗，并促进细胞迁移、侵袭和上皮间质转化（EMT）。但BCAR3在子宫内膜异位症中的表达及其对子宫内膜细胞功能的影响以及子宫内膜异位症的抗雌激素作用尚未见报道。本研究对这些问题进行了解决。方法：本研究纳入32例异位子宫内膜和在位子宫内膜的子宫内膜异位症患者以及31例正常子宫内膜作为对照。通过实时PCR、免疫组化和蛋白质印迹检测BCAR3和微小RNA（miR）-126-5p的表达。采用慢病毒过表达和载体敲低模型、CCK-8实验、Transwell实验以及上皮细胞和间质细胞原代培养的雌激素干预实验验证BCAR3和miR-126-5p对在位子宫内膜细胞形态和生物学行为的影响。结果：BCAR3基因在子宫内膜异位症患者的异位子宫内膜和在位子宫内膜中高表达，且在III-IV期患者优于I-II期患者。体外细胞实验表明，miR-126-5p负向调节BCAR3的表达及其对基质细胞迁移和侵袭的影响。 miR-126-5p的低表达和BCAR3的高表达促进子宫内膜异位症基质细胞的迁移和侵袭。与在位子宫内膜相比，子宫内膜异位症的EMT评估显示，异位子宫内膜中波形蛋白的表达显着增加，而E-钙粘蛋白的表达显着减少。雌激素促进在位子宫内膜上皮细胞的 EMT，而雌激素抑制剂可逆转这种作用。 BCAR3对EMT无直接影响，且不与雌激素协同促进EMT。结论：miR-126-5p负向调控在位子宫内膜异位症中BCAR3的表达，增强子宫内膜细胞的迁移和侵袭，促进子宫内膜异位症的发生。 BCAR3不诱导EMT，与雌激素无协同作用，但其抗雌激素功能的抑制可能为子宫内膜异位症中局部雌激素作用机制提供新的见解。
Purpose: Endometriosis (EMs) is an estrogen-dependent multifactorial disease. Inhibition of estrogen in endometrial cells contributes to their failure to form lesions in ectopic sites. However, whether reducing or suppressing the inhibitory effect of estrogen results in the establishment of ectopic lesions remains unclear. The BCAR3 gene induces estrogen resistance in estrogen-dependent breast cancer cells and promotes cell migration, invasion, and epithelial-mesenchymal transition (EMT). However, the expression of BCAR3 in endometriosis and its effect on endometrial cell function and the anti-estrogen effect of endometriosis have not been reported. These issues are addressed in the present study.Methods: The study included 32 cases of ectopic endometrium and eutopic endometrium in patients with endometriosis and 31 cases of normal endometrium as controls. The expression of BCAR3 and microRNA (miR)-126-5p was detected by real-time PCR, immunohistochemistry, and western blotting. The effects of BCAR3 and miR-126-5p on the morphology and biological behavior of eutopic endometrial cells were verified using lentivirus overexpression and a vector knockdown model, the CCK-8 assay, Transwell experiments, and estrogen intervention experiments using primary cultures of epithelial and stromal cells.Results: The BCAR3 gene was highly expressed in ectopic endometrium and the eutopic endometrium of patients with endometriosis, and the expression level was higher in stage III-IV patients than in stage I-II patients. In vitro cell experiments showed that miR-126-5p negatively regulated the expression of BCAR3 and its effect on the migration and invasion of stromal cells. Low expression of miR-126-5p and high expression of BCAR3 promoted endometriosis stromal cell migration and invasion. Assessment of EMT in endometriosis compared with eutopic endometrium showed that the expression of vimentin was significantly increased and the expression of E-cadherin was significantly decreased in ectopic endometrium. Estrogen promoted EMT in eutopic endometrial epithelial cells and this effect was reversed by estrogen inhibitors. BCAR3 had no direct effect on EMT and did not act synergistically with estrogen on promoting EMT.Conclusion: miR-126-5p negatively regulated BCAR3 expression in eutopic endometriosis, enhanced the migration and invasion of endometrial cells, and promoted the occurrence of endometriosis. BCAR3 did not induce EMT and had no synergistic effect with estrogen, but its inhibition of anti-estrogen function may provide new insight into the mechanism of local estrogen action in endometriosis.