Xu Hai, Sun Yuan, Ma Zhen, Xu Xin, Qin Lili, Luo Baoping
Department of Obstetrics and Gynecology, Huangjiahu Hospital of Hubei University of Chinese Medicine, Wuhan 430065, Hubei, China.
College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, Hubei, China.
Aging (Albany NY). 2018 Nov 28;10(11):3353-3370. doi: 10.18632/aging.101644.
To investigate possible mechanism of abnormal methylation of long non-coding RNA (lncRNA) on endometrial carcinoma (EC) progression, we detected the genome methylation profiling of endometrial carcinoma by bioinformatic analysis. Accordingly, gene was chosen for the further research. We also found that was the target gene of and miRNA, hsa-miR-196a-5p, might form a connection between and . The relationship was further proved by dual-luciferase reporter assay. studies, DNA methylation and expression were determined by MSP and qRT-PCR respectively. Cell proliferation, apoptosis and cell cycle were demonstrated by colony formation assay, Annexin V/PI double staining and flow cytometry. Besides, the function of and in EC was further confirmed by Tumor Xenograft. Our results indicated that EC progression was promoted by hypermethylated and . Moreover, transcription was regulated by via hsa-miR-196a-5p binding. and demethylation by 5-Aza-2-Deoxycytidine inhibited EC cells proliferation and accelerated cell apoptosis. Furthermore, the expression of TACR1, TACR2 and TACR3 was remarkably decreased through and treatments. Our findings establish a novel regulatory axis, /hsa-miR-196a-5p/ Downregulation of the axis promoted EC development through TACR3, which further activated neuroactive ligand-receptor interaction.
为了研究长链非编码RNA（lncRNA）异常甲基化对子宫内膜癌（EC）进展的可能机制，我们通过生物信息学分析检测了子宫内膜癌的基因组甲基化谱。据此，选择了基因进行进一步研究。我们还发现基因是基因的靶基因，且微小RNA hsa-miR-196a-5p可能在基因和基因之间形成联系。双荧光素酶报告基因检测进一步证实了这种关系。通过甲基化特异性PCR（MSP）和定量逆转录聚合酶链反应（qRT-PCR）分别测定研究中的DNA甲基化和表达。通过集落形成试验、膜联蛋白V/碘化丙啶（Annexin V/PI）双染和流式细胞术证明细胞增殖、凋亡和细胞周期。此外，通过肿瘤异种移植进一步证实了基因和基因在子宫内膜癌中的功能。我们的结果表明，基因和基因的高甲基化促进了子宫内膜癌的进展。此外，基因转录通过与hsa-miR-196a-5p结合受到调控。5-氮杂-2'-脱氧胞苷对基因和基因的去甲基化抑制了子宫内膜癌细胞的增殖并加速了细胞凋亡。此外，通过基因和基因处理，速激肽受体1（TACR1）、速激肽受体2（TACR2）和速激肽受体3（TACR3）的表达显著降低。我们的研究结果建立了一个新的调控轴，即基因/hsa-miR-196a-5p/。该轴的下调通过TACR3促进了子宫内膜癌的发展，进而激活了神经活性配体-受体相互作用。
