[ 摘 要 ] 目的：探讨沉默 G 蛋白偶联受体激酶 3（G protein-coupled receptor kinase 3，GRK3）对口腔鳞状细胞癌（oral squamous cell carcinoma，OSCC）细胞增殖、迁移和侵袭的影响及其可能的机制。方法：利用 Oncomine 数据库分析 GRK3 在正 常口腔组织及 OSCC 组织中的表达水平。用 RNA 干扰技术敲降 GRK3 在 OSCC 细胞 WSU-HN6 和 CAL27 中的表达，用 qPCR 法验证干扰效率后，采用 CCK-8 法和流式细胞术分别检测敲降 GRK3 对 OSCC 细胞增殖和凋亡的影响，Transwell 小室 法检测对 OSCC 细胞迁移、侵袭能力的影响，qPCR 法检测对 OSCC 细胞周期、上皮间质转化（epithelial to mesenchymal transition，EMT）和基质金属蛋白酶（matrix metallopeptidase，MMP）相关分子 mRNA 水平表达的影响，WB 法检测 EMT 及 MMP 相关分子的蛋白表达水平变化。结果：OSCC 组织中 GRK3 的表达水平显著高于正常口腔组织（P<0.01）。转染 si-GRK3 后，OSCC 细胞中 GRK3 mRNA 表达水平均下调 70% 以上。敲降 GRK3 可显著抑制 OSCC 细胞的增殖、迁移和侵袭能力（均 P<0.01），对细胞凋亡无显著影响（P>0.05）。敲降 GRK3 表达后，OSCC 细胞的 G0/G1 期比例显著增高（P<0.01），细胞周期蛋白 D1（Cyclin D1）、Cyclin D3、细胞周期蛋白依赖性激酶 2（cyclin-dependent kinases 2，CDK2）和 CDK4 基因的 mRNA 表达降低 （均 P<0.05）；EMT 相关分子波形蛋白（Vimentin）、Zeb1 和 Slug 表达降低，E-钙黏蛋白（E-cadherin）表达升高（均 P<0.05）； MMP3 和 MMP9 表达降低（均 P<0.05），MMP2 和 MMP7 表达无明显变化（均 P>0.05）。结论：GRK3 可通过调节细胞周期促 进 OSCC 细胞的增殖能力，并通过调控 EMT 和 MMP 增强细胞的迁移和侵袭能力。

[Abstract] Objective: To investigate the effects of silencing G protein-coupled receptor kinase 3 (GRK3) on the proliferation, migration and invasion of oral squamous cell carcinoma (OSCC) cells and the possible underlying mechanisms. Methods: GRK3 expression levels in normal oral tissues and OSCC tissues were analyzed using Oncomine database. RNA interference technology was used to down-regulate GRK3 expression in OSCC cell lines WSU-HN6 and CAL27. The interference efficiency was verified by qPCR. The effects of knockdown of GRK3 on proliferation and apoptosis of OSCC cells were detected by CCK-8 assay and Flow cytometry, respectively; the effects on migration and invasion abilities of OSCC cells were determined by Transwell assay; and the effects on mRNA expression levels of molecules associated with cell cycle, epithelial-mesenchymal transition (EMT), and matrix metallopeptidase (MMP) were determined by qPCR. The changes in protein levels of molecules associated with EMT and MMP were detected using WB assay. Results: GRK3 expression level in OSCC tissues was significantly higher than that in normal oral tissues ( P<0.01). After being transfected with si-GRK3, the mRNA expression of GRK3 in OSCC cells was down-regulated by more than 70%. Silencing GRK3 significantly inhibited the proliferation, migration and invasion of OSCC cells (all P<0.01), but had no significant effect on apoptosis (P>0.05). After down-regulation of GRK3, the percentage of OSCC cells in G0/G1 phase was significantly increased (P<0.01); the mRNA expression levels of Cyclin D1, Cyclin D3, CDK2 and CDK4 were decreased (all P<0.05); expression levels of EMT-related proteins (Vimentin, Zeb1 and Slug) were decreased, while E-cadherin was increased (all P<0.05); MMP3 and MMP9 were decreased (all P<0.05), while MMP2 and MMP7 showed no significant changes (all P>0.05). Conclusion: GRK3 promotes the proliferation of OSCC cells by regulating cell cycle-related molecules and enhances the migration and invasion through regulating EMT and MMPs.

上海市“医苑新星”青年医学人才培养计划：青年医学人才类?临床检验项目资助（No. 沪卫人事 [2020] 087 号）；上海交通大学医学院附属第九人民医院院级基金?基础研究助推计划资助（No. JYZZ040）