p53转录调控靶基因GGT6对人结肠癌的影响

doi:10.16352/j.issn.1001-6325.2023.06.0953

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

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.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 Wenjing^1,2, LI Li^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/Abstract

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

中图分类号:

柳子朝, 杜文静, 李莉. p53转录调控靶基因GGT6对人结肠癌的影响[J]. 基础医学与临床, 2023, 43(6): 953-959.

LIU Zizhao, DU Wenjing, LI Li. Effect of p53 transcriptional regulatory target gene GGT6 in human colon adenocarcinoma[J]. Basic & Clinical Medicine, 2023, 43(6): 953-959.

参考文献 11

[1]	Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2021, 71: 209-249.
[2]	Lahalle A, Lacroix M, De Blasio C, et al. The p53 pathway and metabolism: the tree that hides the forest[J]. Cancers (Basel), 2021, 13. doi: 10.3390/cancers13010133.
[3]	Zhao J, Zhou X, Chen B, et al. p53 promotes peroxisomal fatty acid beta-oxidation to repress purine biosynthesis and mediate tumor suppression[J]. Cell Death Dis, 2023, 14. doi: 10.1038/s41419-023-05625-2.
[4]	Rizzotto D, Zaccara S, Rossi A, et al. Nutlin-induced apoptosis is specified by a translation program regulated by PCBP2 and DHX30[J]. Cell Rep, 2020, 30: 4355-4369.
[5]	Heisterkamp N, Groffen J, Warburton D, et al. The human gamma-glutamyltransferase gene family[J]. Hum Genet, 2008, 123: 321-332.
[6]	Reddy RB, Khora SS, Suresh A. Molecular prognos-ticators in clinically and pathologically distinct cohorts of head and neck squamous cell carcinoma-A meta-analysis approach[J]. PLoS One, 2019, 14. doi: 10.1371/journal.pone.0218989.
[7]	Liu Z, Wan Y, Yang M, et al. Identification of methylation-driven genes related to the prognosis of papillary renal cell carcinoma: a study based on The Cancer Genome Atlas[J]. Cancer Cell Int, 2020, 20. doi: 10.1186/s12935-020-01331-7.
[8]	Liu Y, Gu W. The complexity of p53-mediated metabolic regulation in tumor suppression[J]. Semin Cancer Biol, 2022, 85: 4-32.
[9]	Hassin O, Oren M. Drugging p53 in cancer: one protein, many targets[J]. Nat Rev Drug Discov, 2023, 22: 127-144.
[10]	Mottaghitalab F, Lanjanian H, Masoudi-Nejad A. Revealing transcriptional and post-transcriptional regulatory mechanisms of gamma-glutamyl transferase and keratin isoforms as novel cooperative biomarkers in low-grade glioma and glioblastoma multiforme[J]. Genomics, 2021, 113: 2623-2633.
[11]	Corti A, Belcastro E, Dominici S, et al. The dark side of gamma-glutamyltransferase (GGT): Pathogenic effects of an ‘antioxidant' enzyme[J]. Free Radic Biol Med, 2020, 160: 807-819.

[1]	姚安军, 陈凌子, 金惠仙. 二十二碳六烯酸抑制人结肠癌细胞系HT-29增殖[J]. 基础医学与临床, 2024, 44(8): 1107-1112.
[2]	冯娜欣, 王鹏, 何明璇, 刘锦燕. 茯苓酸抑制人结肠癌细胞系HT-29增殖[J]. 基础医学与临床, 2023, 43(11): 1673-1679.
[3]	鄢雯, 李囡, 古同男, 孔祥照. 从GEO数据库筛选结肠癌差异关键基因及验证[J]. 基础医学与临床, 2023, 43(11): 1685-1692.
[4]	侯欣然, 厚静雅, 张亚茹, 李海丽. 叉头框蛋白M1在脑胶质瘤中作用的研究进展[J]. 基础医学与临床, 2023, 43(10): 1604-1607.
[5]	高洪婷, 黄雨晴, 杨黎星, 马瑞, 王钰铖, 鞠瑞, 郭磊. 羧胺三唑增强小鼠脾脏淋巴细胞促进结肠癌细胞系MC38和C26凋亡[J]. 基础医学与临床, 2021, 41(6): 805-810.
[6]	姚涵, 徐文彬, 王冬来. 抗肿瘤药物FTY720对p53依赖的基因转录调控[J]. 基础医学与临床, 2021, 41(3): 311-317.
[7]	闫晓俊, 徐文彬, 王冬来. 羧基末端乙酰化修饰调控抑癌蛋白p53转录特异性[J]. 基础医学与临床, 2021, 41(11): 1577-1582.
[8]	辛玉, 贺龙梅, 杨红, 吕红, 谭蓓, 汪红英, 钱家鸣. 1α,25-二羟基维生素D₃抑制人结肠癌细胞系SW480中β-catenin转录活性[J]. 基础医学与临床, 2020, 40(9): 1169-1174.
[9]	田春阳, 刘晓政, 范军朝. 奥沙利铂联合柔红霉素诱导结肠癌细胞系HT-29凋亡[J]. 基础医学与临床, 2020, 40(3): 310-314.
[10]	石光, 宁娜, 侯晓鸿, 孙聪, 原媛, 王丽, 王晓晖. Rev-erb激动剂SR9009通过降低自噬抑制人结肠癌细胞系HCT116增殖[J]. 基础医学与临床, 2020, 40(11): 1494-1499.
[11]	敖宁王亮王琼张秀茹. USP22 siRNA通过TIMP-2抑制人结肠癌细胞系SW480侵袭[J]. 基础医学与临床, 2019, 39(3): 375-380.
[12]	吴银芳孙晓东郑宇黄东胜. RNA干扰下调RNF2抑制胰腺癌细胞系PANC-1增殖和迁移[J]. 基础医学与临床, 2018, 38(1): 74-79.
[13]	李明星蒋德旗王艳马艳娇喻珊珊王勇. YC-1对低氧诱导人肺动脉平滑肌细胞增殖和p53表达的影响[J]. 基础医学与临床, 2015, 35(10): 1303-1307.
[14]	肖琳王平董俊红王守训刘顺梅. RNAi双沉默Survivin和hTERT基因抑制人结肠癌SW480细胞增殖促进凋亡[J]. 基础医学与临床, 2015, 35(1): 38-43.
[15]	于晓剑王燕李瑞峰李莉刘玉刚. microRNA-34c调控c-Met抑制肝细胞癌的发生发展[J]. 基础医学与临床, 2014, 34(9): 1250-1254.

p53转录调控靶基因GGT6对人结肠癌的影响

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

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 11

相关文章 15

编辑推荐

Metrics

本文评价