SIRT3在皮肤鳞状细胞癌中的作用OACSTPCD
Role of SIRT3 in skin squamous cell carcinoma
目的 探索去乙酰化酶3(Sirtuin-3,SIRT3)对皮肤鳞状细胞癌的影响及潜在调控机制.方法 采用基因表达谱交互分析数据库(Gene Expression Profiling Interactive Analysis,GEPIA)分析SIRT3在多种肿瘤中的表达.采用SIRT3小干扰RNA转染A431细胞株构建SIRT3敲低细胞(si-SIRT3组),将阴性对照siRNA转染至A431细胞中,构建对照组细胞.采用RT-qPCR及Western blot分别检测细胞中SIRT3 mRNA及蛋白水平,采用流式细胞仪检测细胞凋亡率、活性氧(reactive oxygen species,ROS)及膜电位(mitochondrial membrane potential,MMP)水平,采用Western blot检测细胞中促凋亡蛋白Bax、细胞色素C蛋白及抑凋亡蛋白Bcl-2的表达,采用CCK-8实验检测细胞的增殖.A431细胞分别用SIRT3抑制剂3-TYP处理(3-TYP组)和等量DMSO处理(对照组)后,采用流式细胞仪检测细胞凋亡率.采用转录组测序分析对照组和si-SIRT3组的差异表达基因,并对差异表达基因进行GO及KEGG富集分析.结果 SIRT3在胸腺瘤及弥漫性大B细胞淋巴瘤中表达上调(P＜0.05).相比于对照组,si-SIRT3组细胞凋亡率明显增加(P＜0.01),ROS释放增加(P＜0.05),MMP明显降低(P＜0.001),促凋亡蛋白Bax和细胞色素C蛋白表达上调,抑凋亡蛋白Bcl-2表达下调,细胞增殖受到抑制(P＜0.001).相比于对照组,3-TYP组细胞凋亡率增加(P＜0.001).转录组测序分析结果显示,对照组和si-SIRT3组细胞中差异表达基因共有94个,其中上调基因59个,下调基因35个,差异表达基因主要富集在代谢、信号转导、钙离子、蛋白转运、蛋白水解、胞外基质、金属内肽酶及激素分泌等信号通路中.结论 下调皮肤鳞癌细胞中SIRT3的表达,可以促进癌细胞凋亡,抑制增殖,SIRT3有望作为皮肤鳞癌患者新的治疗靶点.
Objective To investigate the effects of Sirtuin-3(SIRT3)on skin squamous cell carcinoma(SCC)and its potential regula-tory mechanism.Methods The expression of SIRT3 in various tumors was analyzed using the Gene Expression Profiling Interactive Analysis(GEPIA)database.A431 cell line was transfected with SIRT3 small interfering RNA(siRNA)to construct SIRT3 knockdown cells(si-SIRT3 group),and A431 cell line was transfected with negative control siRNA as control group.RT-qPCR and Western blot were used to detect SIRT3 mRNA and protein levels.Flow cytometry was employed to measure the apoptosis rate,the reactive oxygen species(ROS)level,and the mitochondrial membrane potential(MMP).Western blot was also applied to detect the expression of the apoptotic protein Bax,cytochrome C protein,and the anti-apoptotic protein Bcl-2.CCK-8 assay was used to assess the proliferation rate.A431 cells were treated with a SIRT3 inhibitor 3-TYP and an equal amount of DMSO,respectively,and then the apoptosis rate was detected by flow cytometry.Differentially expressed genes between control group and si-SIRT3 group were identified by transcrip-tome sequencing analysis,and analyzed by GO and KEGG enrichment analysis.Results SIRT3 expression was upregulated in thymoma and diffuse large B-cell lymphoma(P＜0.05).Compared with control group,the apoptosis rate was significantly increased in si-SIRT3 group(P＜0.01),ROS release was elevated(P＜0.05),MMP was significantly reduced(P＜0.001),the expressions of Bax and cytochrome C protein were upregulated,the expression of Bcl-2 was downregulated,and the cell proliferation was inhibited(P＜0.001).The apoptosis rate in 3-TYP group was also increased compared to control group(P＜0.001).Transcriptome sequencing analysis revealed 94 differentially expressed genes between control group and si-SIRT3 group,including 59 upregulated genes and 35 downregulated genes.These genes were primarily enriched in signaling pathways related to metabolism,signal transduction,calcium ion regulation,protein transport,proteolysis,extracellular matrix,metallopeptidases,and hormone secretion.Conclusion Downregulation of SIRT3 expression in skin squamous cell carcinoma cells can promote the cell apoptosis,inhibit the cell proliferation,and SIRT3 is expected to be a new therapeutic target for patients with skin squamous cell carcinoma.
魏艳;王丽娟;周艳;郑焱
西安交通大学第一附属医院皮肤科,西安 710061
临床医学
皮肤鳞癌SIRT3活性氧细胞凋亡线粒体膜电位
skin squamous cell carcinomaSIRT3reactive oxygen speciescell apoptosismitochondrial membrane potential
《山西医科大学学报》 2024 (007)
820-827 / 8
国家自然科学基金项目(81972938);陕西省科技重点基金资助项目(2015KTCL03-10);陕西省自然科学基础研究计划项目(2024JC-YBQN-0783);西安交通大学第一附属医院科研发展基金项目(2022QN-12)
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