沉默circ-NDRG1对口腔鳞状细胞癌细胞恶性生物学行为的作用机制研究  

Mechanism of silencing circ-NDRG1 regulating the malignant biological behavior of oral squamous cell carcinoma cells

作　　者：陶冶[1] 肖锋[2] 杨文宇[2] 徐培[1] TAO Ye;XIAO Feng;YANG Wenyu;XU Pei(School of Stomatology,Anhui Medical College,Hefei 230601,Anhui,China)

机构地区：[1]安徽医学高等专科学校口腔医学院,安徽合肥230601 [2]安徽医科大学第二附属医院口腔颌面外科,安徽合肥230601

出　　处：《皖南医学院学报》2024年第3期210-214,共5页Journal of Wannan Medical College

基　　金：安徽高校自然科学研究重点项目(KJ2020A0862);高校优秀青年人才支持项目一般项目(gxyq2022192)。

摘　　要：目的:探究circ-NDRG1在口腔鳞状细胞癌组织中的表达及其通过靶向miR-1271对口腔鳞状细胞癌HSC-3细胞恶性生物学行为的影响。方法:利用TCGA数据库分析口腔鳞状细胞癌组织中circ-NDRG1的表达水平,分析circ-NDRG1表达与患者预后生存期的相关性。实时荧光定量PCR(qPCR)检测circ-NDRG1在永生化口腔角质细胞(HOK)和口腔鳞状细胞癌HSC-3、SCC-25、CAL-27、Tca-8113细胞中的表达水平。利用Lipofectamine 2000将NC inhibitor、circ-NDRG1 inhibitor分别转染HSC-3细胞,分别为Anti-NC组和Anti-circ-NDRG1组。MTT法和Transwell小室实验分别检测各组HSC-3细胞增殖和侵袭能力。Western blot检测各组HSC-3细胞增殖蛋白(CDK3、Cyclin C)和上皮间质转化(EMT)蛋白(ZLF8、Notch、SIX1)表达。利用TCGA数据库分析口腔鳞状细胞癌组织中circ-NDRG1与miR-1271表达的关系。双荧光素酶报告实验验证circ-NDRG1和miR-1271的靶向关系。qPCR检测各组HSC-3细胞中miR-1271的表达。结果:TCGA数据库显示口腔鳞状细胞癌组织中circ-NDRG1的表达高于癌旁组织(P<0.01)。口腔鳞状细胞癌患者预后生存期与circ-NDRG1表达水平呈负相关(P<0.01)。与HOK细胞相比,HSC-3、SCC-25、CAL-27、Tca-8113细胞中circ-NDRG1呈高表达(P<0.01)。与Anti-NC组相比,低表达circ-NDRG1能够抑制HSC-3细胞的增殖能力(P<0.05)和侵袭能力(P<0.01)。与Anti-NC组相比,低表达circ-NDRG1能够下调HSC-3细胞中CDK3、Cyclin C、ZLF8、Notch、SIX1蛋白的表达(P<0.01)。口腔鳞状细胞癌组织中circ-NDRG1表达水平与miR-1271表达水平呈负相关(P<0.01)。circ-NDRG1能够靶向结合miR-1271(P<0.01)。与Anti-NC组相比,低表达circ-NDRG1能够上调HSC-3细胞中miR-1271的表达(P<0.01)。结论:口腔鳞状细胞癌组织中circ-NDRG1呈高表达,沉默circ-NDRG1通过靶向调控miR-1271表达抑制口腔鳞状细胞癌细胞的增殖、侵袭和EMT进程。Objective:To investigate the expression of circ-NDRG1 in oral squamous cell carcinoma(OSCC)tissues and the effect on the malignant biological behavior of OSCC HSC-3 cells by targeting miR-1271.Methods:The TCGA database was used to analyze the expression level of circ-NDRG1 in OSCC tissues,and the correlation between its expression and patient′s survival was analyzed.The expression level of circ-NDRG1 in immortalized oral keratinocyte HOK and OSCC HSC-3,SCC-25,CAL-27,Tca-8113 cells was detected by real-time fluorescence quantitative PCR(qPCR).NC inhibitor and circ-NDRG1 inhibitor were transfected into HSC-3 cells respectively by using Lipofectamine 2000(Anti-NC group and Anti-circ-NDRG1 group).The proliferation and invasion abilities of HSC-3 cells in each group were detected by MTT and Transwell chamber assay.The expression of HSC-3 cell proliferation proteins(CDK3,Cyclin C)and epithelial-mesenchymal transition(EMT)proteins(ZLF8,Notch,SIX1)was detected by Western blot.The TCGA database was used to analyze the relationship between the expression of circ-NDRG1 and miR-1271 in OSCC tissues.The dual-luciferase reporter assay was used to verify the targeting relationship between circ-NDRG1 and miR-1271.The expression of miR-1271 in HSC-3 cells in each group was detected by qPCR.Results:TCGA database analysis showed that the expression of circ-NDRG1 in OSCC tissues was significantly higher than that in adjacent tissues(P<0.01).The survival time of OSCC patients was negatively correlated with the expression level of circ-NDRG1(P<0.01).Compared with HOK cells,circ-NDRG1 was highly expressed in HSC-3,SCC-25,CAL-27,and Tca-8113 cells(P<0.01).Compared with the Anti-NC group,low expression of circ-NDRG1 could inhibit the proliferation ability(P<0.05)and invasion ability(P<0.01)of HSC-3 cells.Compared with the Anti-NC group,low expression of circ-NDRG1 could downregulate the protein expressions of CDK3,Cyclin C,ZLF8,Notch,and SIX1 in HSC-3 cells(all P<0.01).The expression level of circ-NDRG1 in OSCC tissues was negatively c

关 键 词：口腔鳞状细胞癌 circ-NDRG1 miR-1271 增殖 侵袭 上皮间质转化 

分 类 号：R739.8[医药卫生—肿瘤]