p53正向调控TIMP2并抑制结直肠癌细胞恶性表型OA
p53 Positively Regulates TIMP2 and Suppresses Malignant Phenotypes of Colorectal Cancer Cells
目的 探讨 p53 对金属蛋白酶组织抑制因子2(tissue inhibitor of metalloproteinases 2,TIMP2)的转录调控作用及其对结直肠癌细胞生物学功能的作用.方法 采用 p53 缺失 HCT116 细胞转录组数据筛选差异表达基因;选取 HCT116 与 HCT8 两类结直肠癌细胞株,在 p53 敲降及激活条件下利用 qRT-PCR 与蛋白质印迹测定 p53 对 TIMP2 的转录调控效应;结合 JASPAR 数据库与双荧光素酶报告实验,验证 p53 直接转录调控 TIMP2;利用划痕和 Transwell 实验检测 p53 过表达与 TIMP2 缺失对肿瘤细胞迁移和侵袭的作用;利用 TCGA 数据库分析结直肠癌(TCGA-COAD)中癌与癌旁组织中 TIMP2 的表达并进行生存情况分析.结果 RNA-seq 分析显示 TIMP2 为差异表达较显著的基因之一;在 HCT116 与 HCT8 细胞中 TIMP2 mRNA和蛋白质水平均与 p53 呈正相关;p53 可直接转录激活 TIMP2,进而抑制肿瘤的恶性表型.TCGA 数据库显示 COAD 肿瘤中 TIMP2 低表达,且与结直肠癌患者预后较差相关.结论 TIMP2 在结直肠癌细胞中低表达,p53 部分通过转录调控 TIMP2 抑制肿瘤细胞转移,表明 TIMP2 有望成为未来治疗结直肠癌的潜在靶点之一.
Objective To explore the transcriptional regulatory effect of p53 on tissue inhibitor of metalloproteinases 2(TIMP2)and its role in the biological functions of colorectal cancer cells.Methods Differentially expressed genes were screened using transcriptome data of p53-defi-cient HCT116 cells.Two colorectal cancer cell lines,HCT116 and HCT8,were selected,and the transcriptional regulatory effect of p53 on TIMP2 was measured by qRT-PCR and Western blotting under p53 knockdown and activation conditions.The direct transcriptional regulation of TIMP2 by p53 was verified via combining the JASPAR database and dual-luciferase reporter assay.The cell scratch and Transwell assays were used to detect the effects of p53 overexpression and TIMP2 deple-tion on tumor cell migration and invasion.The TCGA database was used to analyze TIMP2 expression and survival in cancer and adjacent normal tissues in colorectal cancer(TCGA-COAD).Results RNA-seq analysis showed TIMP2 was among the significantly differentially expressed genes.In HCT116 and HCT8 cells,TIMP2 mRNA and protein levels were positively correlated with p53.p53 could directly activate TIMP2 transcription,thereby suppressing tumor malignant phenotypes.The TCGA database showed low TIMP2 expression in COAD tumors,which was associated with poor prognosis in colorectal cancer patients.Conclusion TIMP2 is lowly expressed in colorectal cancer cells,and p53 partially inhibits tumor cell metastasis by transcriptionally regulating TIMP2,indica-ting that TIMP2 is expected to become a potential target for the future treatment of colorectal cancer.
袁嘉阳;杜文静;李薇
中国医学科学院北京协和医学院基础医学研究所细胞生物学系 北京市,100005中国医学科学院北京协和医学院基础医学研究所细胞生物学系 北京市,100005||重大疾病共性机制研究全国重点实验室 北京市,100005中国医学科学院北京协和医学院基础医学研究所细胞生物学系 北京市,100005
生物科学
p53金属蛋白酶组织抑制因子2结直肠癌转录调控侵袭
p53tissue inhibitor of metalloproteinases 2colorectal cancertranscriptional regulationinvasion
《医学分子生物学杂志》 2026 (3)
233-241,9
国家自然科学基金(No.82303260),中国医学科学院医学与健康科技创新工程(No.2025-I2M-XHJC-018、No.2025-I2M-XHXX-066) This work was supported by grants from the National Natural Science Foundation of China(No.82303260)and the Medical and Health Technology Inno-vation Project of the Chinese Academy of Medical Sciences(No.2025-I2M-XHJC-018,No.2025-I2M-XHXX-066).
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