TGF-β1通过诱导代谢重组促进子宫内膜异位症内膜间质细胞的迁移和侵袭

Metabolic reprogramming induced by transforming growth factor-beta 1 promotes the migration and invasion of endometrial stromal cells

目的:探讨转化生长因子β1(TGF-β1)是否具有诱导子宫内膜间质细胞发生代谢重组的效应,及该效应对内膜间质细胞生物学行为的影响及其可能作用机制。方法:收集子宫内膜异位症(EMs)患者的异位病灶及在位内膜。免疫组化法分析标本组织内无氧糖酵解标志分子单羧酸转运蛋白1和4(MCT1、MCT4)等表达。体外分离和培养获得原代内膜间质细胞。原代培养的内膜间质细胞用TGF-β1(2ng/ml)干预12h,检测培养液中乳酸浓度。siRNA沉默MCT4基因表达。qRT-PCR和Western blot法检测无氧糖酵解相关基因和蛋白的表达水平,Transwell小室检测细胞迁移和侵袭能力。结果:内异症异位病灶组织内无氧糖酵解标志分子MCT1、MCT4表达显著增加。相较无干预细胞组,外源性TGF-β1作用后内膜间质细胞无氧糖酵解标志分子MCT1、MCT4和乳酸脱氢酶A(LDHA)表达均显著上调(P均<0.05);同时TGF-β1干预促进内膜间质细胞的乳酸分泌及迁移和侵袭(P均<0.05),但MCT4基因沉默可显著逆转上述效应。结论:TGF-β1通过诱导内膜间质细胞无氧糖酵解、乳酸分泌促进了细胞的迁移和侵袭,且MCT4在其中发挥了关键性作用。

Objective: To investigate whether TGF-β1 induces the metabolic reprogramming in endometrial stromal cells (ESCs) and then affects the biological behavior of ESCs.To further explore the possible mechanism. Methods: Ectopic and eutopic endometria were collected from patients with endometriosis.The expression of glycolysis-associated markers,monocarboxylate transporter 1 and 4,were assessed by immunohistochemical staining.Primary human ESCs were isolated and cultured in vitro from eutopic endometria of IVF-ET patients with endometriosis.After ESCs were treated with 2ng/ml TGF-β1 for 12 hours,the concentration of lactic acid in the condition media was detected.Cells were transfected with siRNA to knock down the MCT4 expression.The mRNA and protein expressions of MCT1,MCT4 and lactate dehydrogenase A (LDHA) were analyzed by qRT-PCR and Western blot respectively.Cell migration and invasion were evaluated by Transwell assay. Results: The expressions of glycolysis-associated markers MCT1 and MCT4 significantly increased in the ectopic lesions.Compared to the control group (without TGF-β1 stimulation),exposure of ESCs to TGF-β1 significantly increased the expressions of LDHA mRNA and MCT1/4 mRNA and proteins (all P < 0.05 ).In addition,TGF-β1 stimulation promoted the lactic acid production,migration and invasion of endometrial stromal cells (all P <0.05).While MCT4 silencing in the ESCs significantly reversed the above effects. Conclusion: TGF-β1 promotes the migration and invasion of ESCs by inducing glycolysis and lactate secretion,in which MCT4 plays a pivotal role.