The 2nd Dental Center, Ninth People,s Hospital, Shanghai Jiao Tong University School of Medicine; National Clinical Research Center for Oral Disease; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai 200011, China.
Dept. of Oral and Maxillofacial-Head and Neck Oncology, Ninth People,s Hospital, Shanghai Jiao Tong University, School of Medicine; National Clinical Research Center for Oral Disease; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai 200011, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2021 Jun 1;39(3):320-327. doi: 10.7518/hxkq.2021.03.012.
This study aims to investigate the effect of the regulator of G-protein signaling 2 (RGS2) on the proliferation and invasion of oral squamous cell carcinoma (OSCC) cells and its potential molecular mechanism. Metho⁃ds The expression status and clinical significance of RGS2 in head and neck squamous cell carcinomas and matched adjacent normal tissues were evaluated using TCGA database. Three OSCC cell lines (i.e., SCC-9, Cal27, and Fadu) were overexpressed with RGS2, and the effect of RGS2 on cell proliferation and invasion was determined using the Transwell, clone formation, and cell counting kit (CCK)-8 assays. Moreover, the yeast two-hybrid scree-ning and co-immunoprecipitation (Co-IP) assays were conducted to detect the correlation of RGS2, four and a half LIM domains protein 1 (FHL1), and damage DNA-binding protein 1 (DDB1).
The expression level of RGS2 in OSCC was significantly lower than that in matched adjacent normal tissues (=0.023). The high RGS2 expression level was negatively correlated with lymphovascular invasion (<0.001). After transfection with lentiv-RGS2, the expression of RGS2 was increased, and the invasion and proliferation abilities of OSCC cell lines were evidently inhibited. FHL1 could competitively bind with RGS2, which decreased the integration of DDB1 and RGS2, inhibited the ubiquitination process of RGS2, and maintained the stability of the RGS2 protein.
RGS2 plays an important role in the inhibition of OSCC proliferation and invasion. The structure stability of RGS2 is competitively regulated by FHL1 and DDB1.
本研究旨在探讨 G 蛋白信号调节因子 2（RGS2）对口腔鳞状细胞癌（OSCC）细胞增殖和侵袭的影响及其潜在的分子机制。
利用 TCGA 数据库评估 RGS2 在头颈部鳞状细胞癌及其配对的正常组织中的表达状态和临床意义。采用 RGS2 过表达的三种 OSCC 细胞系（即 SCC-9、Cal27 和 Fadu），通过 Transwell、克隆形成和细胞计数试剂盒（CCK-8）检测 RGS2 对细胞增殖和侵袭的影响。此外，进行酵母双杂交筛选和免疫共沉淀（Co-IP）实验，以检测 RGS2、四个半 LIM 结构域蛋白 1（FHL1）和损伤 DNA 结合蛋白 1（DDB1）之间的相关性。
OSCC 中 RGS2 的表达水平明显低于配对的正常组织（=0.023）。高 RGS2 表达水平与血管淋巴管浸润呈负相关（<0.001）。转染慢病毒-RGS2 后，RGS2 的表达增加，OSCC 细胞系的侵袭和增殖能力明显受到抑制。FHL1 可以与 RGS2 竞争结合，从而减少 DDB1 和 RGS2 的整合，抑制 RGS2 的泛素化过程，并维持 RGS2 蛋白的稳定性。
RGS2 在抑制 OSCC 增殖和侵袭中发挥重要作用。FHL1 和 DDB1 竞争性调节 RGS2 的结构稳定性。
