Department of Otorhinolaryngology, Head and Neck Surgery, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, Guangdong, China.
Guangdong Provincial Key Laboratory of Biomedical Imaging, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, Guangdong, China.
PeerJ. 2022 Mar 30;10:e13221. doi: 10.7717/peerj.13221. eCollection 2022.
Long non-coding RNA SPRY4 intronic transcript 1 (Lnc RNA SPRY4-IT1) was aberrant-expressed in various kinds of cancer. Increasing evidence demonstrated that lnc RNAs involved in tumorigenesis and metastasis. In this study, we aimed to explore the biological role of SPRY4-IT1 on the phenotype of nasopharyngeal carcinoma (NPC) in vitro and in vivo.
The expression level of SPRY4-IT1 in NPC cell lines were measured by quantitative real-time polymerase chain reaction (qRT-PCR). Cell Counting Kit-8 (CCK-8) and colony formation assay were used to detect cell proliferation. Wound-healing assay, transwell assay and animal experiment were performed to evaluate the ability of cell migration and metastasis. Cell cycle distribution and apoptosis were determined by flow cytometry. Western blotting and immunofluorescence were employed to identify protein expression.
SPRY4-IT1 was significantly up-regulated in several NPC cell lines (6-10B, CNE-2, and HONE-1) compared with human immortalized nasopharyngeal epithelial cell (NP69). Silencing of SPRY4-IT1 inhibited proliferation, migration, and metastasis, and induced significant G2/M phase arrest and apoptosis. Western blotting showed that the expression levels of cell cycle-related proteins (cyclin B1, cdc2 and p-cdc2) were down-regulated and apoptosis-associated proteins (PARP, cleaved PARP and cleaved caspase-3) were up-regulated after knockdown of SPRY4-IT1. The expression level of E-cadherin was increased and the expression of Vimentin, Snail and Twist1 were decreased after the SPRY4-IT1 knockdown.
lncRNA SPRY4-IT1 played a significant role in NPC proliferation, migration and metastasis, suggesting that SPRY4-IT1 might be a potential therapeutic target for the treatment of NPC.
长链非编码 RNA SPRY4 内含子转录本 1（Lnc RNA SPRY4-IT1）在各种癌症中异常表达。越来越多的证据表明，lncRNA 参与肿瘤的发生和转移。在这项研究中，我们旨在探讨 SPRY4-IT1 对体外和体内鼻咽癌（NPC）表型的生物学作用。
通过实时定量聚合酶链反应（qRT-PCR）测量 NPC 细胞系中 SPRY4-IT1 的表达水平。使用细胞计数试剂盒-8（CCK-8）和集落形成实验检测细胞增殖。通过划痕愈合实验、Transwell 实验和动物实验评估细胞迁移和转移能力。通过流式细胞术检测细胞周期分布和细胞凋亡。采用 Western blot 和免疫荧光法鉴定蛋白表达。
与永生化鼻咽上皮细胞（NP69）相比，几种 NPC 细胞系（6-10B、CNE-2 和 HONE-1）中 SPRY4-IT1 显著上调。沉默 SPRY4-IT1 抑制增殖、迁移和转移，并诱导显著的 G2/M 期阻滞和细胞凋亡。Western blot 显示，沉默 SPRY4-IT1 后细胞周期相关蛋白（cyclin B1、cdc2 和 p-cdc2）的表达水平下调，凋亡相关蛋白（PARP、cleaved PARP 和 cleaved caspase-3）的表达水平上调。沉默 SPRY4-IT1 后 E-钙黏蛋白表达增加，波形蛋白、Snail 和 Twist1 的表达减少。
lncRNA SPRY4-IT1 在 NPC 的增殖、迁移和转移中发挥重要作用，提示 SPRY4-IT1 可能是 NPC 治疗的潜在靶点。
