Department of Gynecology, The Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou 221000, Jiangsu, China.
Department of Gynecology, The Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou 221000, Jiangsu, China.
Reprod Biol. 2022 Jun;22(2):100648. doi: 10.1016/j.repbio.2022.100648. Epub 2022 May 6.
Cervical cancer (CC) is a common gynecological malignant tumor, causing poor survival rate. Circular RNAs (circRNAs) are abundantly expressed in CC with their stable loop structure. However, the underlying mechanism and biological function of circRNAs remained unclear. Using quantitative real-time polymerase chain reaction (qRT-PCR) or western blot assay, we measured the expression of hsa_circ_0001495, miR-526b-3p, and transmembrane Bax inhibitor motif containing 6 (TMBIM6) in CC tissues and cells. The relationship between miR-526b-3p and hsa_circ_0001495 or TMBIM6 was investigated by bioinformatics analysis, dual-luciferase and RIP analysis. Enzyme linked immunosorbent assay (ELISA) was conducted to evaluate glucose consumption and lactate production. 5-ethynyl-2'-deoxyuridine (EDU) assay were used to test cell proliferation. Cell apoptosis was analyzed by using flow cytometry assay. Transwell and wound-healing assays were used to measure cell invasion and migration. The expression of proteins was examined by western blot. Xenograft assay was applied to detect the effect of hsa_circ_0001495 in vivo. Our finding showed that hsa_circ_0001495 and TMBIM6 expression were upregulated, while miR-526b-3p was downregulated in CC tissues and cell lines. Hsa_circ_0001495 knockdown or TMBIM6 knockdown suppressed cell proliferation, migration, glycolysis, while promoted cell apoptosis in vitro, and hsa_circ_0001495 silence curbed tumor growth in vivo. Beside, hsa_circ_0001495 exerted its function in CC by positively regulating TMBIM6. Furthermore, hsa_circ_0001495 acted as a sponge for miR-526b-3p to regulate TMBIM6 expression. Hsa_circ_0001495/miR-526b-3p/TMBIM6 axis also regulated the phosphorylation of mammalian target of rapamycin (mTOR) in CC cells. In summary, hsa_circ_0001495 regulated the progression of CC by regulating miR-526b-3p/TMBIM6/mTOR pathway.
宫颈癌(CC)是一种常见的妇科恶性肿瘤,其生存率较差。环状 RNA(circRNA)在 CC 中大量表达,具有稳定的环结构。然而,circRNAs 的潜在机制和生物学功能尚不清楚。我们使用定量实时聚合酶链反应(qRT-PCR)或 Western blot 检测 CC 组织和细胞中 hsa_circ_0001495、miR-526b-3p 和跨膜 Bax 抑制剂基序包含 6(TMBIM6)的表达。通过生物信息学分析、双荧光素酶和 RIP 分析研究 miR-526b-3p 与 hsa_circ_0001495 或 TMBIM6 的关系。酶联免疫吸附试验(ELISA)用于评估葡萄糖消耗和乳酸生成。EDU 测定用于检测细胞增殖。通过流式细胞术分析检测细胞凋亡。Transwell 和划痕愈合试验用于测量细胞侵袭和迁移。Western blot 检测蛋白表达。异种移植试验用于检测 hsa_circ_0001495 在体内的作用。我们的研究结果表明,hsa_circ_0001495 和 TMBIM6 的表达在 CC 组织和细胞系中上调,而 miR-526b-3p 的表达下调。hsa_circ_0001495 敲低或 TMBIM6 敲低抑制细胞增殖、迁移和糖酵解,同时促进细胞凋亡,hsa_circ_0001495 沉默抑制体内肿瘤生长。此外,hsa_circ_0001495 通过正向调节 TMBIM6 发挥其在 CC 中的作用。此外,hsa_circ_0001495 作为 miR-526b-3p 的海绵来调节 TMBIM6 的表达。hsa_circ_0001495/miR-526b-3p/TMBIM6 轴还调节 CC 细胞中哺乳动物雷帕霉素靶蛋白(mTOR)的磷酸化。总之,hsa_circ_0001495 通过调节 miR-526b-3p/TMBIM6/mTOR 通路调节 CC 的进展。