基于NF-κB信号通路探究SIRT7基因对口腔癌细胞株自噬蛋白及巨噬细胞极化的作用机制  

Study on the mechanism of SIRT7 gene on autophagy protein and macrophage polarization of oral cancer cell line based on NF-κB signal pathway

作　　者：顾超[1] 张红梅 仝越越 孙红玲 何晓娉 GU Chao;ZHANG Hong-mei;TONG Yue-yue;SUN Hong-ling;HE Xiao-ping(Zhongda Hospital Affiliated to Southeast University,Jiangsu Nanjing 210044,China)

机构地区：[1]东南大学附属中大医院,江苏南京210044

出　　处：《临床口腔医学杂志》2023年第12期712-717,共6页Journal of Clinical Stomatology

基　　金：国家自然科学基金项目(编号:81502340)。

摘　　要：目的:基于NF-κB信号通路探究SIRT7基因对口腔癌细胞株自噬蛋白及巨噬细胞极化的作用机制。方法:人口腔鳞状细胞癌细胞株SCC25细胞分为SCC25组(无干预的SCC25细胞)、阴性对照组(SCC25细胞转染空载体pLKO.1-vector-GFP)、SIRT7 shRNA组(SCC25细胞转染SIRT7 shRNA)、SIRT7组(SCC25细胞转染SIRT7慢病毒),联合A组(SCC25细胞+50μmol/L NF-κB抑制剂PDTC+SIRT7 shRNA)及联合B组(SCC25细胞+50μmol/L NF-κB抑制剂PDTC+SIRT7慢病毒),qRT-PCR检测SIRT7相对表达;Transwell法检测侵袭数目;划痕实验检测划痕愈合率;免疫荧光检测P62、LC3-Ⅱ表达,免疫印迹分别检测M1和M2标志蛋白及NF-κB、NF-κB p65蛋白水平。结果:SCC25组及阴性对照组SCC25细胞的侵袭数目、划痕愈合率、P62、LC3-Ⅱ、CD163、CD11C、NF-κB及NF-κB p65表达比较差异无统计学意义(P>0.05);与阴性对照组相比,SIRT7 shRNA组细胞侵袭数目增加、划痕愈合率、CD163、NF-κB及NF-κB p65升高,CD11C降低(P<0.05);与SIRT7 shRNA组相比,SIRT7组SCC25细胞的侵袭数目、划痕愈合率、CD163、NF-κB及NF-κB p65降低,CD11C升高(P<0.05)。与联合A组相比,且联合B组SCC25细胞的侵袭数目、划痕愈合率、CD163、NF-κB及NF-κB p65降低,CD11C升高(P<0.05)。结论:上调SIRT7可抑制口腔鳞状细胞癌细胞侵袭及迁移,降低自噬蛋白P62、LC3-Ⅱ活性,促进巨噬细胞M2向M1极化,研究机制可能与SIRT7调节NF-κB相关。Objective:To explore the mechanism of SIRT7 gene on autophagy protein and macrophage polarization in oral squamous cell carcinoma cells based on the NF-κB signaling pathway.Methods:Human oral squamous cell carcinoma cell line SCC25 cells were divided into SCC25 group(SCC25 cells without intervention),negative control group(SCC25 cells transfected with empty vector pLKO.1-vector-GFP),SIRT7 shRNA group(SCC25 cells transfected with SIRT7 shRNA),SIRT7 group(SCC25 cells transfected SIRT7 lentivirus),combination group A(SCC25 cells+50μmol/L NF-κB inhibitor PDTC+SIRT7 shRNA)and combination group B(SCC25 cells+50μmol/L NF-κB inhibitor PDTC+SIRT7 lentivirus),the relative expression of SIRT7 was detected by qRT-PCR.The number of invasion was detected by Transwell method.The scratch healing rate was detected by scratch test.Immunofluorescence detection of P62,LC3-II,the levels of M1 and M2 marker proteins,NF-κB and NF-κB p65 proteins were detected by Western blot.Results:There was no significant difference in SCC25 cells invasion number,scratch healing rate,P62,LC3-II,CD163,CD11C,NF-κB and NF-κB p65 expression between SCC25 group and negative control group(P>0.05).The number of cell invasion,scratch healing rate,CD163,NF-κB and NF-κB p65 were increased and CD11C was decreased in SIRT7 shRNA group(P<0.05).The invasion number,scratch healing rate,CD163,NF-κB and NF-κB p65 of SCC25 cells in SIRT7 group were decreased,while CD11C was increased(P<0.05).Compared with combination group A,the invasion number,scratch healing rate,CD163,NF-κB and NF-κB p65 of SCC25 cells in combination group B were decreased,and CD11C was increased(P<0.05).Conclusion:Up-regulation of SIRT7 can inhibit the invasion and migration of oral squamous cell cancer cells,reduce the activities of autophagy protein P62 and LC3-II,and promote the polarization of macrophage M2 to M1.The mechanism may be related to the regulation of NF-κB by SIRT7.

关 键 词：口腔癌 SIRT7 自噬蛋白 巨噬细胞极化 核转录因子NF-ΚB 

分 类 号：R739.8[医药卫生—肿瘤]