Clinical Laboratory, Urumqi Maternal and Child Health Care Hospital, Urumqi, Xinjiang, China.
Department of Gynecology, Urumqi Maternal and Child Health Care Hospital, Urumqi, Xinjiang, China.
Biochem Genet. 2022 Dec;60(6):2611-2629. doi: 10.1007/s10528-022-10227-2. Epub 2022 Jun 8.
Accumulating evidence have demonstrated that circular RNAs (circRNAs) exert important roles in tumor initiation and progression. Nevertheless, the role and mechanism of circRNA ceramide synthase 6 (circCERS6) in epithelial ovarian cancer (EOC) remain to be clarified. Using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot assay, we measured RNA and protein expression. We analyzed the effects of circCERS6/microRNA-630 (miR-630)/Ras-association domain family member 8 (RASSF8) axis in cell proliferation, migration, and invasion by 5-Ethynyl-2'-deoxyuridine (EdU) assay, colony formation assay, scratch test, and transwell assay. Using RNA-pull down assay and dual-luciferase reporter assay, the interaction between miR-630 and circCERS6 or RASSF8 was verified. The in vivo role of circCERS6 in tumor growth was analyzed using xenograft mice model. CircCERS6 expression was markedly reduced in EOC tissues and cell lines. CircCERS6 overexpression hampered the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of EOC cells. CircCERS6 directly interacted with miR-630. miR-630 expression was up-regulated in EOC tissues and cell lines. CircCERS6 overexpression-induced anti-tumor effects in EOC were largely reversed by the overexpression of miR-630. RASSF8 was a direct target of miR-630. RASSF8 level was decreased in EOC tissues and cell lines. CircCERS6 up-regulated RASSF8 expression by acting as a molecular sponge for miR-630 in EOC cells. CircCERS6 overexpression-mediated suppressive effects in EOC cells were largely overturned by the silence of RASSF8. CircCERS6 overexpression blocked tumor growth in vivo. CircCERS6 overexpression hampered the proliferation, migration, invasion, and EMT of EOC cells by up-regulating RASSF8 via sponging miR-630.
越来越多的证据表明，环状 RNA（circRNA）在肿瘤的发生和发展中发挥着重要作用。然而，环状 RNA 神经酰胺合酶 6（circCERS6）在卵巢上皮性癌（EOC）中的作用和机制仍有待阐明。本研究采用逆转录定量聚合酶链反应（RT-qPCR）和 Western blot 检测，测量 RNA 和蛋白表达。通过 5-乙炔基-2'-脱氧尿苷（EdU）检测、集落形成实验、划痕实验和 Transwell 实验分析 circCERS6/微小 RNA-630（miR-630）/Ras 相关结构域家族成员 8（RASSF8）轴对细胞增殖、迁移和侵袭的影响。采用 RNA 下拉实验和双荧光素酶报告基因实验验证 miR-630 与 circCERS6 或 RASSF8 的相互作用。利用异种移植小鼠模型分析 circCERS6 在体内肿瘤生长中的作用。结果显示，circCERS6 在 EOC 组织和细胞系中表达明显降低。circCERS6 过表达抑制 EOC 细胞的增殖、迁移、侵袭和上皮间质转化（EMT）。circCERS6 与 miR-630 直接相互作用。miR-630 在 EOC 组织和细胞系中表达上调。circCERS6 过表达引起的 EOC 抗肿瘤作用在很大程度上被 miR-630 的过表达所逆转。RASSF8 是 miR-630 的直接靶标。RASSF8 在 EOC 组织和细胞系中表达下调。circCERS6 通过在 EOC 细胞中作为 miR-630 的分子海绵来上调 RASSF8 的表达。circCERS6 过表达介导的 EOC 细胞抑制作用在 RASSF8 沉默时被大大推翻。circCERS6 过表达在体内阻断肿瘤生长。circCERS6 过表达通过海绵吸附 miR-630 上调 RASSF8 来抑制 EOC 细胞的增殖、迁移、侵袭和 EMT。
