G蛋白偶联受体激酶3在口腔鳞状细胞癌细胞增殖、迁移和侵袭中的作用及其可能的机制  

Role of G protein-coupled receptor kinase 3 in the proliferation,migration and invasion of oral squamous carcinoma cells and its possible mechanism

作　　者：张晗 罗清琼[1] 朱丽萍[1] 陈福祥[1] ZHANG Han;LUO Qingqiong;ZHU Liping;CHEN Fuxiang(Department of Clinical Laboratory,Shanghai Ninth People’s Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院检验科,上海200011

出　　处：《中国肿瘤生物治疗杂志》2021年第5期435-442,共8页Chinese Journal of Cancer Biotherapy

基　　金：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)。

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(t=5.799,P<0.01),细胞周期蛋白D1(Cyclin D1)、Cyclin D3、周期蛋白依赖性激酶2(cyclin-dependent kinases 2,CDK2)和CDK4基因的m RNA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。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(t=5.799,P<0.01);the mRNA expression levels of Cyclin D1,Cyclin D3,CDK2 and CDK4 were decreased(all P<0.05);expression 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 promotes the migration and invasion through regulating EMT and MMPs.

关 键 词：口腔鳞状细胞癌 G蛋白偶联受体激酶3 增殖 迁移 侵袭 上皮-间质转化 基质金属蛋白酶 

分 类 号：R739.85[医药卫生—肿瘤] R730.2[医药卫生—临床医学]