乳腺癌抑癌基因KRT17过表达通过活化p53信号通路促进癌细胞凋亡并阻滞细胞周期  

Overexpression of breast cancer suppressor gene KRT17 promotes apoptosis and arrests cell cycle by activating p53 signaling pathway

作　　者：陈铎 胡明[1] 周海瑞[1] 张雪松[1] CHEN Duo;HU Ming;ZHOU Hairui;ZHANG Xuesong(Plastic Surgery Department,School of Clinical Medicine,Jiamusi University,Heilongjiang Jiamusi 154007,China)

机构地区：[1]佳木斯大学临床医学院整形外科,黑龙江佳木斯154007

出　　处：《现代肿瘤医学》2024年第17期3180-3189,共10页Journal of Modern Oncology

基　　金：黑龙江省自然科学基金项目(编号:LH2021H104)。

摘　　要：目的:探讨乳腺癌中角蛋白17(keratin 17,KRT17)的表达、预后意义及其对癌细胞生长的影响与机制。方法:基于癌症基因组图谱(The Cancer Genome Atlas,TCGA)收录的乳腺癌(breast cancer,BC)数据筛选差异表达基因和预后相关基因,取交集获得靶基因KRT17。分别利用UALCAN、人类蛋白质图谱(Human Protein Mapping,HPA)和TISCH2数据库分析各种癌症中KRT17 mRNA和蛋白水平的表达、BC组织芯片中KRT17的表达与定位以及肿瘤微环境中KRT17的表达,利用bc-GenExMiner v5.0数据库分析不同数据集中KRT17表达与预后的相关性。接下来,检测KRT17过表达对MDA-MB-231和MCF7细胞生长的影响,并用全基因组敲除和敲低文库筛选数据验证KRT17对BC细胞系生长的影响,裸鼠移植瘤实验验证KRT17对裸鼠异种移植瘤生长的调节作用。最后,利用流式细胞术检测过表达KRT17对细胞周期和凋亡的调控,GSEA富集分析KRT17功能,RT-qPCR和Western blotting揭示KRT17促癌生长的分子机制。结果:不同肿瘤中KRT17表达具有异质性;BC中KRT17mRNA和蛋白水平显著下调(P<0.001,P<0.001),KRT17蛋白主要定位于癌细胞的细胞质和细胞膜,且肿瘤微环境中各种细胞均表达KRT17;KRT17低表达是BC预后不良因素(均P<0.01),过表达KRT17可以显著抑制MDA-MB-231和MCF7细胞的生长(P<0.01,P<0.01),敲除或敲低KRT17可促进MDA-MB-231和MCF7等多种BC细胞系的生长,裸鼠异体移植瘤模型证实过表达KRT17可抑制肿瘤体积(P<0.001);过表达KRT17可显著促进细胞凋亡(P <0.001)并阻滞细胞周期G1期(P <0.01);分子实验结果显示p53信号通路部分介导了KRT17过表达对细胞凋亡和周期的调节作用。结论:KRT17在BC中低表达,不利于患者预后,过表达KRT17通过活化p53通路促进细胞凋亡并阻滞细胞周期,KRT17是一种潜在的抑癌基因。Objective:To investigate the expression and prognostic significance of keratin 17(KRT17) in breast cancer(BC) and its effect on the growth of cancer cells and its mechanism.Methods:Based on the BC data collected by the Cancer Genome Atlas(TCGA),differential expression genes and prognostic related genes were screened,and the target gene KRT17 was obtained by intersection.UALCAN,Human Protein Mapping(HPA) and TISCH2 databases were used to analyze the levels of KRT17 mRNA and protein in various cancers,the expression and localization of KRT17 in BC tissues,and the expression of KRT17 in tumor microenvironment,respectively.The correlation between KRT17 expression and prognosis in different datasets was also analyzed using the bc-GenExMiner v5.0 database.Next,the effects of KRT17 overexpression on the growth of MDA-MB-231 and MCF7 cells were examined,and the effects of KRT17 on the growth of BC cell lines were verified by genome-wide knockout and knockdown library screening data,and the regulatory effects of KRT17 on the growth of xenografts in nude mice were verified in vivo.Finally,flow cytometry was used to detect the regulation of overexpression of KRT17 on cell cycle and apoptosis,and the function of KRT17 was analyzed by GSEA enrichment.RT-qPCR and Western blotting were used to reveal the molecular mechanism of KRT17 promoting cancer growth.Results:The expression of KRT17 was heterogeneous in different tumors.The mRNA and protein levels of KRT17 in BC were significantly down-regulated(P<0.001,P<0.001).KRT17 protein was mainly located in the cytoplasm and cell membrane of cancer cells,and KRT17 was expressed in various cells in the tumor microenvironment.Low expression of KRT17 was a poor prognostic factor for BC(all P<0.01),and overexpression of KRT17 could significantly inhibit the growth of MDA-MB-231 and MCF7 cells(P<0.01,P<0.01).Knockout or knockdown of KRT17 promoted the growth of multiple BC cell lines such as MDA-MB-231 and MCF7,and overexpression of KRT17 inhibited tumor volume in nude mouse xenografts(P

关 键 词：乳腺癌 角蛋白17 预后 p53信号通路 细胞生长 

分 类 号：R737.9[医药卫生—肿瘤]