Department of Obstetrics and Gynecology, Tongji Hospital Affiliated to Tongji University, Shanghai, China.
Clin Exp Pharmacol Physiol. 2021 Jun;48(6):890-901. doi: 10.1111/1440-1681.13456. Epub 2021 Feb 28.
Database screening indicated that tubulin polymerization-promoting protein 3 (TPPP3) was involved in pathogenesis of multiple cancer types. miR-1827 has a potential role in a variety of human cancers. However, the role of TPPP3 and its underlying molecular mechanism in endometrial cancer (EC) has not been investigated. Herein, we aimed to reveal the role of TPPP3/miR-1827 in EC progression.
Tumour tissue and whole blood samples were collected for the detection of TPPP3 expression. TPPP3 shRNAs and pcDNA-TPPP3 were applied to knockdown or upregulate the TPPP3 expression, and miR-1827 mimic was used to upregulate miR-1827 level. CCK-8 and colony assays were applied to estimate the cell proliferation. Wound healing and Transwell assays were conducted to assess the cell migration and invasion abilities. The dual-luciferase reporter assay was conducted to validate the putative binding site between TPPP3 and miR-1827. Expression of TPPP3, miR-1827 and related proteins in cell lines, tissue and whole blood sample were detected using western blot, RT-qPCR and immunofluorescence.
TPPP3 was observed markedly elevated in EC patients and cells. TPPP3 knockdown displayed evident suppression in cell proliferation, migration and invasion in vitro and in vivo. Moreover, we identified TPPP3 as a direct and functional target gene of miR-1827 in EC cells. The miR-1827 induced regulatory effects on EC cells were partially reversed by TPPP3. Additionally, in vivo study confirmed the findings discovered in vitro.
TPPP3 exerted oncogenic roles in EC progression by sponging miR-1827. This finding might provide potential targets for EC therapy.
数据库筛选表明，微管蛋白聚合促进蛋白 3（TPPP3）参与了多种癌症的发病机制。miR-1827 在多种人类癌症中具有潜在作用。然而，TPPP3 在子宫内膜癌（EC）中的作用及其潜在的分子机制尚未得到研究。在此，我们旨在揭示 TPPP3/miR-1827 在 EC 进展中的作用。
收集肿瘤组织和全血样本以检测 TPPP3 表达。应用 TPPP3 shRNAs 和 pcDNA-TPPP3 下调或上调 TPPP3 表达，并用 miR-1827 模拟物上调 miR-1827 水平。CCK-8 和集落形成实验用于评估细胞增殖。伤口愈合和 Transwell 实验用于评估细胞迁移和侵袭能力。双荧光素酶报告实验用于验证 TPPP3 和 miR-1827 之间的假定结合位点。使用 Western blot、RT-qPCR 和免疫荧光法检测细胞系、组织和全血样本中 TPPP3、miR-1827 和相关蛋白的表达。
TPPP3 在 EC 患者和细胞中明显升高。TPPP3 下调在体外和体内显著抑制细胞增殖、迁移和侵袭。此外，我们鉴定出 TPPP3 是 EC 细胞中 miR-1827 的直接和功能靶基因。TPPP3 部分逆转了 miR-1827 对 EC 细胞的诱导调节作用。此外，体内研究证实了体外发现的结果。
TPPP3 通过海绵吸附 miR-1827 在 EC 进展中发挥致癌作用。这一发现可能为 EC 治疗提供潜在的靶点。
