Chen Xubin, Chen Qiaojiang, Zhao Chen, Lu Zhiqi
Department of Oral and Maxillofacial Surgery, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, China.
Department of Anesthesiology, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, China.
J Dent Sci. 2023 Jan;18(1):282-294. doi: 10.1016/j.jds.2022.05.012. Epub 2022 Jun 4.
BACKGROUND/PURPOSE: Circular RNAs (circRNAs) have been identified as potential functional modulators of the cellular physiology processes. This study aims to learn the potential molecular mechanisms of hsa_circ_0005050 (circ_0005050) in oral squamous cell carcinoma (OSCC).
Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to examine the expression of circ_0005050, miR-487a-3p, and chondroitin sulfate synthase 1 (CHSY1). Dual-luciferase reporter system, RNA pull-down, and RNA Immunoprecipitation (RIP) assays were used to determine the binding between miR-487a-3p and circ_0005050 or CHSY1. Colony formation experiment and EdU assay were used to investigate proliferation. Wound-healing and transwell assays were used to detect the migration of cells. The apoptosis rate of OSCC cells was tested by flow cytometry. Protein levels of related factors were determined by Western blot. Tumor xenograft was established to determine the regulatory role of circ_0005050 on tumor growth , and Ki-67 expression was detected in this xenograft using Immunohistochemical (IHC).
We implicated that circ_0005050 was apparently upregulated in OSCC tissues cells. In function experiments, repressing of circ_0005050 remarkably retarded OSCC growth . Furthermore, we conducted dual-luciferase reporter assays and RNA pull-down assays to verify that circ_0005050 sponged miR-487a-3p. Suppression of miR-487a-3p rescued the inhibition of proliferation in SCC15 and SCC25 cells induced by circ_0005050 knockdown. In addition, we found that overexpression of CHSY1 also reversed the inhibitory effect of circ_0005050 silencing on cell proliferation. Moreover, circ_0005050 knockdown inhibited tumor growth .
Circ_0005050 acted as an oncogenic factor in OSCC progression through miR-487a-3p/CHSY1 axis.
背景/目的：环状RNA（circRNAs）已被确定为细胞生理过程的潜在功能调节因子。本研究旨在了解hsa_circ_0005050（circ_0005050）在口腔鳞状细胞癌（OSCC）中的潜在分子机制。
采用定量逆转录-聚合酶链反应（qRT-PCR）检测circ_0005050、miR-487a-3p和硫酸软骨素合酶1（CHSY1）的表达。利用双荧光素酶报告系统、RNA下拉和RNA免疫沉淀（RIP）实验来确定miR-487a-3p与circ_0005050或CHSY1之间的结合。采用集落形成实验和EdU实验研究增殖情况。采用伤口愈合实验和Transwell实验检测细胞迁移。通过流式细胞术检测OSCC细胞的凋亡率。用蛋白质印迹法测定相关因子的蛋白水平。建立肿瘤异种移植模型以确定circ_0005050对肿瘤生长的调节作用，并使用免疫组织化学（IHC）检测该异种移植模型中Ki-67的表达。
我们发现circ_0005050在OSCC组织细胞中明显上调。在功能实验中，抑制circ_0005050显著抑制了OSCC的生长。此外，我们进行了双荧光素酶报告实验和RNA下拉实验，以验证circ_0005050可吸附miR-487a-3p。抑制miR-487a-3p可挽救circ_0005050敲低诱导的SCC15和SCC25细胞增殖抑制。此外，我们发现CHSY1的过表达也可逆转circ_0005050沉默对细胞增殖的抑制作用。此外，circ_0005050敲低抑制了肿瘤生长。
Circ_0005050通过miR-487a-3p/CHSY1轴在OSCC进展中发挥致癌因子的作用。
