目的:Src同源磷酸酪氨酸磷酸酶2(SHP2)与几种癌症类型的进展有关。然而,其在子宫内膜癌(EC)中的功能尚不清楚。这里,我们报道了十11易位3(TET3)介导的DNA去甲基化修饰负责SHP2在EC中的致癌作用,并探讨了其详细机制。
方法:使用生物信息学工具分析EC组织和对照组织之间的转录组差异,其次是蛋白质-蛋白质相互作用网络的建立。用靶向SHP2的shRNA单独处理EC细胞,或与异孕酮醇联合处理,表皮生长因子受体(EGFR)信号激活因子。使用细胞计数试剂盒-8检查细胞生物学行为,集落形成,流式细胞术,划痕试验,和transwell分析,并计算了醋酸甲羟孕酮/吉非替尼的中位抑制浓度值.证实了TET3与SHP2启动子的结合。选择具有TET3敲低并与SHP2过表达组合的EC细胞以在小鼠中构建肿瘤异种移植物。
结果:TET3和SHP2在EC细胞中过表达。TET3与SHP2启动子结合,从而增加DNA羟甲基化修饰并激活SHP2以诱导EGFR/细胞外信号调节激酶(ERK)途径。TET3或SHP2的敲除抑制EC细胞恶性侵袭性并损害EGFR/ERK途径。TET3的沉默抑制了EC细胞的致瘤能力,SHP2或异孕酮烯醇的异位表达逆转了TET3敲低对EC细胞生物学活性的抑制作用。
结论:TET3促进SHP2启动子的DNA去甲基化修饰并激活SHP2,从而激活EGFR/ERK通路并导致EC进展。
OBJECTIVE: Src homology phosphotyrosin phosphatase 2 (SHP2) has been implicated in the progression of several cancer types. However, its function in endometrial cancer (EC) remains unclear. Here, we report that the ten-eleven translocation 3 (TET3)-mediated DNA demethylation modification is responsible for the oncogenic role of SHP2 in EC and explore the detailed mechanism.
METHODS: The transcriptomic differences between EC tissues and control tissues were analyzed using bioinformatics tools, followed by protein-protein interaction network establishment. EC cells were treated with shRNA targeting SHP2 alone or in combination with isoprocurcumenol, an epidermal growth factor receptor (EGFR) signaling activator. The cell biological behavior was examined using cell counting kit-8, colony formation, flow cytometry, scratch assay, and transwell assays, and the median inhibition concentration values to medroxyprogesterone acetate/gefitinib were calculated. The binding of TET3 to the SHP2 promoter was verified. EC cells with TET3 knockdown and combined with SHP2 overexpression were selected to construct tumor xenografts in mice.
RESULTS: TET3 and SHP2 were overexpressed in EC cells. TET3 bound to the SHP2 promoter, thereby increasing the DNA hydroxymethylation modification and activating SHP2 to induce the EGFR/extracellular signal-regulated kinase (ERK) pathway. Knockdown of TET3 or SHP2 inhibited EC cell malignant aggressiveness and impaired the EGFR/ERK pathway. Silencing of TET3 inhibited the tumorigenic capacity of EC cells, and ectopic expression of SHP2 or isoprocurcumenol reversed the inhibitory effect of TET3 knockdown on the biological activity of EC cells.
CONCLUSIONS: TET3 promoted the DNA demethylation modification in the SHP2 promoter and activated SHP2, thus activating the EGFR/ERK pathway and leading to EC progression.