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目的研究瘦素(Leptin)对HTR8-SVneo滋养细胞系增殖的影响及对DNA结合和分化抑制剂(ID)分子表达水平的影响,探讨Leptin对HTR8-SVneo细胞增殖作用的调节机制。方法体外培养HTR8-SVneo滋养细胞,MTT法检测Leptin不同质量浓度(0、10、50、100、250、500 ng/mL)刺激后细胞增殖情况。HTR8-SVneo细胞培养体系中加入Leptin(0、100、500 ng/mL),RT-PCR检测瘦素受体(Leptin R)各亚型表达,实时定量PCR检测ID分子表达,Western Blot检测ID2表达。SiRNA干扰ID2分子后,分为Leptin(0 ng/mL)、Leptin(100 ng/mL)、Leptin(500 ng/mL)、Leptin(0 ng/mL)+ID2-siRNA、Leptin(100 ng/mL)+ID2-siRNA、Leptin(500 ng/mL)+ID2-siRNA,MTT检测Leptin对HTR8-SVneo滋养细胞增殖的影响。结果与Leptin(0 ng/mL)比较,高质量浓度Leptin(100、250、500 ng/mL)促进HTR8-SVneo细胞增殖(P<0.05);HTR8-SVneo细胞表达除HuB219.2外的全部受体亚型;Leptin以剂量依赖方式促进HTR8-SVneo细胞ID2分子及OB-RL表达(P<0.05);ID2-siRNA抑制ID2表达并逆转Leptin对滋养细胞增殖的促进作用。结论 Leptin可能通过Leptin R-ID2途径促进HTR8-SVneo细胞增殖,为阐明滋养细胞增殖调控机制提供了新的基础数据。
Objective To investigate the effect of Leptin on the proliferation of HTR8-SVneo trophoblast cell line and the effect of Leptin on the molecular expression of DNA binding and differentiation inhibitor (ID), and to explore the regulatory mechanism of Leptin on HTR8-SVneo cell proliferation. Methods The HTR8-SVneo trophoblast cells were cultured in vitro. The proliferation of HTR8-SVneo cells was detected by MTT assay. The proliferation of HTR8-SVneo cells was detected by MTT assay at different concentrations of Leptin (0, 10, 50, 100, 250, 500 ng / mL). Leptin (0, 100, 500 ng / mL) was added into HTR8-SVneo cell culture system. The expression of Leptin R subtype was detected by RT-PCR. The expression of ID was detected by real- . After siRNA interfered with ID2, Leptin (100 ng / mL), Leptin (500 ng / mL), Leptin (0 ng / mL) ) + ID2-siRNA, Leptin (500 ng / mL) + ID2-siRNA were used to detect the effect of Leptin on the proliferation of HTR8-SVneo trophoblast cells. Results Compared with Leptin (0 ng / mL), high concentration of Leptin (100, 250 and 500 ng / mL) promoted HTR8-SVneo cell proliferation (P <0.05) Leptin promoted the expression of ID2 and OB-RL in HTR8-SVneo cells in a dose-dependent manner (P <0.05). ID2-siRNA suppressed the expression of ID2 and reversed the promoting effect of Leptin on the proliferation of trophoblasts. Conclusion Leptin may promote the proliferation of HTR8-SVneo cells through the Leptin R-ID2 pathway, providing new basic data for elucidating the regulatory mechanism of trophoblast proliferation.