Department of Women'ss Health Service, Yantaishan Hospital, YanTai, Shandong, China.
Department of Gynecology, Yantaishan Hospital, No. 91 Jiefang Road, Zhifu District, YanTai, 264000, Shandong, China.
J Mol Histol. 2021 Apr;52(2):245-256. doi: 10.1007/s10735-020-09944-y. Epub 2021 Jan 4.
Long non-coding RNA (lncRNA) has been demonstrated as vital regulator in human cancer. However, the precise role of lnc-TDRG1 in cervical cancer (CC) remains unclear, so this study was aimed to clarify the role and underlying molecular mechanism of lnc-TDRG1 in CC. The real-time quantitative polymerase chain reaction (RT-qPCR) was conducted to assess the expression levels of lnc-TDRG1, miR-214-5p and Semaphorin 4C (SEMA4C). Under hypoxia condition, the biological behaviors of CC cell, including invasion and glycolysis were determined by transwell assay and Glucose Assay Kit and Lactate Assay Kit, respectively. The Western blot assay was employed to test the expression level of SEMA4C and hexokinase 2 (HK2) expression. The interaction relationship between miR-214-5p and lnc-TDRG1 or SEMA4C was analyzed bioinformatics database and confirmed by dual-luciferase reporter assay, respectively. A xenograft experiment in nude mice was established to clarify the functional role of lnc-TDRG1 in vivo. We found Lnc-TDRG1 was highly expressed in CC tissues and cells and it was upregulated in response to hypoxia. Loss-of-functional experiment suggested that knockdown of lnc-TDRG1 impede invasion, hypoxia-induced glycolysis in vitro and tumor growth in vivo, which was abolished by knockdown of miR-214-5p or overexpression of SEMA4C. Moreover, we confirmed that miR-214-5p specifically bound to SEMA4C and negatively correlated with SEMA4C expression. Collectively, lnc-TDRG1 regulated SEMA4C expression by sponging miR-214-5p in CC. Collectively, mechanistically, lnc-TDRG1 could act as a sponge of miR-214-5p to regulate the expression of SEMA4C, and further regulate invasion and hypoxia-glycolysis in CC cells.
长链非编码 RNA（lncRNA）已被证明是人类癌症的重要调节因子。然而，lnc-TDRG1 在宫颈癌（CC）中的确切作用尚不清楚，因此本研究旨在阐明 lnc-TDRG1 在 CC 中的作用及其潜在的分子机制。实时定量聚合酶链反应（RT-qPCR）用于评估 lnc-TDRG1、miR-214-5p 和 Semaphorin 4C（SEMA4C）的表达水平。在低氧条件下，通过 Transwell 测定和葡萄糖测定试剂盒和乳酸测定试剂盒分别测定 CC 细胞的生物学行为，包括侵袭和糖酵解。Western blot 测定用于检测 SEMA4C 和己糖激酶 2（HK2）表达水平。通过生物信息学数据库分析 miR-214-5p 与 lnc-TDRG1 或 SEMA4C 的相互作用关系，并分别通过双荧光素酶报告基因实验进行验证。建立裸鼠移植瘤实验以阐明 lnc-TDRG1 在体内的功能作用。我们发现 Lnc-TDRG1 在 CC 组织和细胞中高表达，并且对低氧有反应性上调。功能丧失实验表明，敲低 lnc-TDRG1 可阻止体外侵袭、低氧诱导的糖酵解和体内肿瘤生长，而敲低 miR-214-5p 或过表达 SEMA4C 则可消除这种作用。此外，我们证实 miR-214-5p 特异性结合 SEMA4C 并与 SEMA4C 表达呈负相关。总之，lnc-TDRG1 通过海绵吸附 miR-214-5p 调节 CC 中的 SEMA4C 表达。总之，机制上，lnc-TDRG1 可以作为 miR-214-5p 的海绵，调节 SEMA4C 的表达，进而调节 CC 细胞的侵袭和低氧糖酵解。
