METHODS: Tumor and adjacent normal tissues were collected from cervical cancer patients, and the expression of EBLN3P and miR-29c-3p were analyzed via RT-qPCR. The capacities of proliferation, migration, and invasion were assessed using CCK-8, wound healing and transwell assays. The interaction among EBLN3P, miR-29c-3p and TAF15 was determined by luciferase, RNA immunoprecipitation and RNA pull-down assays, respectively. A subcutaneous tumor xenograft mouse model was established to evaluate the functional role of EBLN3P in vivo.
RESULTS: The interaction and reciprocal negative regulation between EBLN3P and miR-29c-3p were uncovered in cervical cancer cells. Likewise, EBLN3P and miR-29c-3p expression patterns in tumor tissues presented a negative association. EBLN3P knockdown weakened cell proliferation, migration and invasion, but these effects were abrogated by miR-29c-3p depletion. Mechanistically, ALKBH5 might impaired EBLN3P stability to reduce its expression. EBLN3P functioned as a competing endogenous RNA (ceRNA) for miR-29c-3p to relieve its suppression of RCC2. Besides, EBLN3P enhanced RCC2 mRNA stability via interacting with TAF15. Furthermore, silencing of EBLN3P repressed the tumor growth in mice.
CONCLUSIONS: Altogether, lncRNA EBLN3P positively regulates RCC2 expression via competitively binding to miR-29c-3p and interacting with TAF15, thereby boosting proliferation, migration, and invasion of cervical cancer cells.
方法:收集宫颈癌患者肿瘤及癌旁正常组织,通过RT-qPCR分析EBLN3P和miR-29c-3p的表达。扩散能力,迁移,使用CCK-8,伤口愈合和transwell测定评估和侵袭。EBLN3P之间的相互作用,通过荧光素酶测定miR-29c-3p和TAF15,RNA免疫沉淀和RNA下拉法,分别。建立皮下肿瘤异种移植小鼠模型以评估EBLN3P在体内的功能作用。
结果:在宫颈癌细胞中发现了EBLN3P与miR-29c-3p之间的相互作用和相互负调控。同样,EBLN3P和miR-29c-3p在肿瘤组织中的表达模式呈现负相关。EBLN3P敲低削弱细胞增殖,移民和入侵,但这些影响被miR-29c-3p耗尽所消除。机械上,ALKBH5可能会损害EBLN3P的稳定性以降低其表达。EBLN3P充当miR-29c-3p的竞争性内源性RNA(ceRNA)以减轻其对RCC2的抑制。此外,EBLN3P通过与TAF15相互作用增强RCC2mRNA稳定性。此外,EBLN3P的沉默抑制了小鼠的肿瘤生长。
结论:总而言之,lncRNAEBLN3P通过竞争性结合miR-29c-3p并与TAF15相互作用,从而促进增殖,迁移,和宫颈癌细胞的侵袭。