Department of Otorhinolaryngology, Weifang Traditional Chinese Medicine Hospital, Weifang, Shandong 261041, PR China.
Department of Infectious Diseases, Weifang people's Hospital, Weifang, Shandong 261041, PR China.
Exp Mol Pathol. 2021 Oct;122:104671. doi: 10.1016/j.yexmp.2021.104671. Epub 2021 Aug 4.
This study aims to investigate the role of lncRNA RHPN1-AS1 in NPC and its potential regulatory mechanism. The expression of RHPN1-AS1 in tissues and cells was measured by qRT-PCR. The effect of RHPN1-AS1 silencing on biological functions of NPC cells was detected by CCK-8, colony formation, flow cytometry, wound healing, and transwell assays. The protein expression was measured by western blot. The RBPs of RHPN1-AS1 were predicted by Starbase and LncTar, and verify by RIP assay. ESTIMATE was used to analyze the relationship between CELF2 expression and tumor purity. GSEA was used to analyze the downstream signaling pathway of CELF2. In our study, RHPN1-AS1 was up-regulated in NPC tissues and cells. RHPN1-AS1 silencing inhibited cell viability, capacity of proliferation, migration and invasion, promoted apoptosis, decreased protein expression of Bcl-2, MMP2/9, increased protein expression of Bax, caspase-3, and TIMP2 of NPC cells. CELF2 was a target of RHPN1-AS1 and was down-regulated in NPC tissues and cells. CELF2 level was associated with tumor purity negatively. Low expression of CELF2 activated mTORC1 signaling pathway and increased protein expression of p-mTORC1/mTORC1 and p-P70S6K/P70S6K. RHPN1-AS1 silencing eliminated the activating effect of CELF2 silencing on mTORC1 signaling pathway. Moreover, CELF2 silencing reversed the inhibitory effect of RHPN1-AS1 on NPC progression. In conclusion, our findings indicated that RHPN1-AS1 plays an important role in NPC via activating mTORC1 signaling which is modulated by CELF2. RHPN1-AS1 may serve as a potential therapeutic target for NPC treatment.
本研究旨在探讨长链非编码 RNA RHPN1-AS1 在 NPC 中的作用及其潜在的调控机制。通过 qRT-PCR 测定组织和细胞中 RHPN1-AS1 的表达。通过 CCK-8、集落形成、流式细胞术、划痕愈合和 Transwell 测定检测 RHPN1-AS1 沉默对 NPC 细胞生物学功能的影响。通过 Western blot 测定蛋白表达。通过 Starbase 和 LncTar 预测 RHPN1-AS1 的 RBPs，并通过 RIP 测定验证。使用 ESTIMATE 分析 CELF2 表达与肿瘤纯度的关系。使用 GSEA 分析 CELF2 的下游信号通路。在本研究中，RHPN1-AS1 在 NPC 组织和细胞中上调。RHPN1-AS1 沉默抑制 NPC 细胞活力、增殖能力、迁移和侵袭能力，促进凋亡，降低 Bcl-2、MMP2/9 蛋白表达，增加 Bax、caspase-3 和 TIMP2 蛋白表达。CELF2 是 RHPN1-AS1 的靶点，在 NPC 组织和细胞中下调。CELF2 水平与肿瘤纯度呈负相关。CELF2 低表达激活 mTORC1 信号通路，增加 p-mTORC1/mTORC1 和 p-P70S6K/P70S6K 蛋白表达。RHPN1-AS1 沉默消除了 CELF2 沉默对 mTORC1 信号通路的激活作用。此外，CELF2 沉默逆转了 RHPN1-AS1 对 NPC 进展的抑制作用。总之，我们的研究结果表明，RHPN1-AS1 通过激活 mTORC1 信号通路在 NPC 中发挥重要作用，该信号通路受 CELF2 调节。RHPN1-AS1 可能成为 NPC 治疗的潜在治疗靶点。
