Circ_0000140 restrains the proliferation, metastasis and glycolysis metabolism of oral squamous cell carcinoma through upregulating CDC73 via sponging miR-182-5p
Circ_0000140 restrains the proliferation, metastasis and glycolysis metabolism of oral squamous cell carcinoma through upregulating CDC73 via sponging miR-182-5p
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DOI：
10.1186/s12935-020-01501-7
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发表时间：
2020-08-26
影响因子：
5.8
通讯作者：
Zhang, Meng
中科院分区：
文献类型：
--
作者：
Guo, Jia;Su, Yuanyuan;Zhang, Meng
背景口腔鳞状细胞癌（OSCC）是世界上较常见的癌症。新的证据表明环状 RNA (circRNA) 参与 OSCC 的进展。然而，circ_0000140在OSCC中的作用仍不清楚。方法通过实时定量聚合酶链式反应（qRT-PCR）评估circ_0000140和microRNA-182-5p（miR-182-5p）的表达。此外，分别通过集落形成和跨孔实验测量细胞增殖、迁移和侵袭。采用蛋白质印迹（WB）分析检测增殖、转移和糖酵解代谢相关蛋白以及细胞分裂周期73（CDC73）蛋白的水平。进一步利用Seahorse XF细胞外通量分析仪检测细胞的细胞外酸化率（ECAR）。采用乳酸测定试剂盒检测细胞的乳酸水平。此外，使用双荧光素酶报告基因验证miR-182-3p与circ_0000140或CDC73之间的相互作用，并采用RNA免疫沉淀（RIP）测定进一步证实miR-182-3p与circ_0000140之间的关系。此外，还建立了小鼠异种移植模型来测量circ_0000140对体内OSCC肿瘤生长的影响。结果 Circ_0000140在OSCC中低表达，其过表达阻碍OSCC细胞的增殖、迁移、侵袭和糖酵解代谢。 MiR-182-5p 可以被 circ_0000140 吸收，其模拟物可以逆转 circ_0000140 过表达对 OSCC 进展的抑制。 CDC73可以被miR-182-3p靶向，其沉默可以逆转miR-182-3p抑制剂对OSCC进展的抑制。此外，过表达的circ_0000140减少了体内OSCC肿瘤的生长。结论 Circ_0000140可能在OSCC中发挥抗癌作用，为OSCC的临床治疗提供了新的靶点。
Background Oral squamous cell carcinoma (OSCC) is a more common cancer in the world. Emerging evidence suggests that circular RNAs (circRNAs) participate in the progression of OSCC. However, the role of circ_0000140 in OSCC is still unknown. Methods The expression of circ_0000140 and microRNA-182-5p (miR-182-5p) were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). Also, cell proliferation, migration and invasion were measured by colony formation and transwell assays, respectively. Western blot (WB) analysis was used to test the levels of proliferation, metastasis and glycolysis metabolism-related proteins as well as cell division cycle 73 (CDC73) protein. Further, the extracellular acidification rate (ECAR) of cells was detected by the Seahorse XF Extracellular Flux Analyzer. The lactate acid level of cells was tested by Lactate Assay Kit. Moreover, dual-luciferase reporter was used to verify the interaction between miR-182-3p and circ_0000140 or CDC73, and RNA immunoprecipitation (RIP) assay was employed to further confirm the relationship between miR-182-3p and circ_0000140. In addition, mice xenograft models were built to measure the effect of circ_0000140 on OSCC tumor growth in vivo. Results Circ_0000140 was lowly expressed in OSCC, and its overexpression hindered proliferation, migration, invasion and glycolysis metabolism in OSCC cells. MiR-182-5p could be sponged by circ_0000140, and its mimic could invert the suppression of circ_0000140 overexpression on OSCC progression. CDC73 could be targeted by miR-182-3p, and its silencing could reverse the inhibition of miR-182-3p inhibitor on OSCC progression. Further, overexpressed circ_0000140 reduced the OSCC tumor growth in vivo. Conclusions Circ_0000140 might play an anti-cancer role in OSCC, which provided a novel target for clinical therapy of OSCC.