基础医学与临床 ›› 2023, Vol. 43 ›› Issue (6): 953-959.doi: 10.16352/j.issn.1001-6325.2023.06.0953

• 研究论文 • 上一篇    下一篇

p53转录调控靶基因GGT6对人结肠癌的影响

柳子朝1, 杜文静1,2, 李莉1*   

  1. 1.中国医学科学院基础医学研究所 北京协和医学院基础学院 细胞生物学系 医学分子生物学国家重点实验室,北京 100005;
    2.山西医科大学 基础医学院 细胞生理学教育部重点实验室,山西 太原 030606
  • 收稿日期:2023-03-07 修回日期:2023-04-18 出版日期:2023-06-05 发布日期:2023-05-31
  • 通讯作者: *rene_ll@126.com
  • 基金资助:
    国家重点研发计划(2019YFA0802600);中国医学科学院医学与健康科技创新工程 (2021-I2M-1-016);细胞生态海河实验室项目(22HHXBSS00011);中国医学科学院医学与健康科技创新工程健康长寿先导科技专项(2019-RC-HL-007)

Effect of p53 transcriptional regulatory target gene GGT6 in human colon adenocarcinoma

LIU Zizhao1, DU Wenjing1,2, LI Li1*   

  1. 1. Skate Key Laboratory of Medical Molecular Biology, Department of Cell Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005;
    2. Key Laboratory of Cell Physiology, Ministry of Education, College of Basic Medicine, Shanxi Medical University, Taiyuan 030606, China
  • Received:2023-03-07 Revised:2023-04-18 Online:2023-06-05 Published:2023-05-31
  • Contact: *rene_ll@126.com

摘要: 目的 筛选出p53转录调控靶基因,并利用生物信息学分析该基因在结肠癌(COAD)中的作用。方法 对p53蛋白稳定剂Nutlin处理的结肠癌细胞系HCT116的RNA-seq数据进行基因差异分析及GO、KEGG和GSEA富集分析;在HCT116及HCT8细胞中加入Nutlin稳定或转染siRNA敲降p53的表达,利用实时荧光定量PCR检测候选靶基因的mRNA表达;双荧光素酶报告实验验证p53对靶基因的转录调控;利用TCGA数据库中的结肠癌数据分析靶基因在结肠癌组织中的表达及患者预后情况。结果 Nutlin处理结肠癌细胞系HCT116后细胞周期、DNA复制及染色体分离等生物过程基因表达发生变化,同样p53信号通路也被激活。在HCT116及HCT8细胞中γ-谷氨酰转移酶(GGT6)的mRNA水平与p53蛋白表达呈正相关。双荧光素酶报告实验确定了GGT6是p53转录调控的靶基因。TCGA-COAD数据分析表明GGT6在结肠癌组织中表达高于癌旁组织(P<0.001),且GGT6高表达患者的总体生存时间较GGT6低表达患者更长(P<0.01),推测GGT6是结肠癌的抑癌因素。结论 GGT6是一个p53正调控转录靶基因,在结肠癌中可能是一个抑癌因素。

关键词: p53, 转录调控, γ-谷氨酰转移酶, 结肠癌

Abstract: Objective To screen out a transcriptional target gene of p53 regulation, and to analyze the role of this gene in colon adenocarcinoma(COAD) by bioinformatics.Methods RNA-seq data of colon cancer cell line HCT116 treated with p53 protein stabilizer Nutlin were analyzed for differential gene expression (DEGs) and GO, KEGG and GSEA enrichment analysis. Nutlin was treated to stabilize p53 or transfected siRNA to knock down p53 expression in HCT116 and HCT8 cells. Quantitative real-time PCR was used to detect the mRNA expression of the predicted target gene. Dual luciferase reporter assay was used to verify the transcriptional regulation of p53 on the target gene. The expression of the target gene and the prognosis of patients was analyzed by TCGA-COAD database. Results After Nutlin treatment, the genes coding for cell cycle, DNA replication and chromosome segregation etc were changed. The p53 signaling pathway was also activated. The mRNA level of GGT6 was positively correlated with the protein expression of p53 in HCT116 and HCT8 cells. Dual luciferase reporter assay identified GGT6 as a target gene of p53 transcriptional regulation. The TCGA-COAD data analysis showed that the expression of GGT6 in COAD tissues was higher than that in the adjacent tissues(P<0.001), and the overall survival(OS) time of patients with high GGT6 expression was longer than that of patients with low GGT6 expression(P<0.01), suggesting GGT6 as a tumor suppressor in COAD. Conclusions GGT6 is a new transcriptional target gene of p53 and a potential tumor suppressor for colon adenocarcinoma development.

Key words: p53, transcriptional regulation, GGT6, colon adenocarcinoma

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