Zhang Cuiying, Liu Wei, Li Fei, Feng Yang, Li Yunyun, Wang Jia
Department of Gynaecology, Yongchuan Hospital of Chongqing Medical University, Chongqing, People's Republic of China.
Department of Orthopedics, Yongchuan Hospital of Chongqing Medical University, Chongqing, People's Republic of China.
Cancer Manag Res. 2021 Mar 10;13:2265-2277. doi: 10.2147/CMAR.S291218. eCollection 2021.
More and more evidences show that circular RNA (circRNA) has an important role in ovarian cancer (OC). Hsa_circ_0015326 is a newly discovered upregulated circRNA in OC, but its role and mechanism in OC have not been studied yet.
Quantitative real-time PCR was used to detect the expression of hsa_circ_0015326, microRNA (miR)-127-3p and MYB. The viability, colony number, cell cycle process, invasion, migration and apoptosis of cells were determined using cell counting kit 8 assay, colony formation assay, flow cytometry, transwell assay and wound healing assay. Moreover, the protein expression levels of metastasis, proliferation, apoptosis markers and MYB were assessed using Western blot analysis. The interaction between miR-127-3p and hsa_circ_0015326 or MYB was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation assay. Xenograft tumors were built to explore the role of hsa_circ_0015326 in OC tumor growth in vivo.
Elevated expression of hsa_circ_0015326 was identified in OC tissues and cells. Loss-of-function experiments suggested that silenced hsa_circ_0015326 inhibited the proliferation, invasion, migration, and promoted the apoptosis of OC cells in vitro, as well as inhibited OC tumorigenesis in vivo. Mechanically, hsa_circ_0015326 sponged miR-127-3p and miR-127-3p targeted MYB. The rescue experiments revealed that miR-127-3p inhibitor reversed the inhibitory effect of hsa_circ_0015326 silencing on OC progression, and MYB overexpression reversed the suppressive effect of miR-127-3p on OC progression. In addition, our data indicated that MYB expression was positively regulated by hsa_circ_0015326.
This study showed that hsa_circ_0015326 could facilitate OC progression by regulating the miR-127-3p/MYB axis, which suggested that it might become a potential target for the treatment of OC.
越来越多的证据表明，环状RNA（circRNA）在卵巢癌（OC）中发挥重要作用。Hsa_circ_0015326是一种新发现的在OC中上调的circRNA，但其在OC中的作用和机制尚未得到研究。
采用定量实时PCR检测hsa_circ_0015326、微小RNA（miR）-127-3p和MYB的表达。使用细胞计数试剂盒8检测、集落形成检测、流式细胞术、Transwell检测和伤口愈合检测来确定细胞的活力、集落数量、细胞周期进程、侵袭、迁移和凋亡。此外，使用蛋白质印迹分析评估转移、增殖、凋亡标志物和MYB的蛋白质表达水平。通过双荧光素酶报告基因检测和RNA免疫沉淀检测证实miR-127-3p与hsa_circ_0015326之间的相互作用。构建异种移植肿瘤以探讨hsa_circ_0015326在体内OC肿瘤生长中的作用。
在OC组织和细胞中鉴定出hsa_circ_0015326表达升高。功能丧失实验表明，沉默hsa_circ_0015326可抑制OC细胞在体外的增殖、侵袭、迁移，并促进其凋亡，以及在体内抑制OC肿瘤发生。机制上，hsa_circ_0015326吸附miR-127-3p，且miR-127-3p靶向MYB。挽救实验表明，miR-127-3p抑制剂可逆转hsa_circ_0015326沉默对OC进展的抑制作用，而MYB过表达可逆转miR-127-3p对OC进展的抑制作用。此外，我们的数据表明hsa_circ_0015326对MYB表达具有正向调节作用。
本研究表明，hsa_circ_0015326可通过调节miR-127-3p/MYB轴促进OC进展，这表明它可能成为OC治疗的潜在靶点。
