泛素特异性蛋白酶20、低氧诱导因子1α在乳腺癌中的表达变化及意义  

Expression and significance of ubiquitin ⁃ specific proteases 20 and hypoxia inducible factor ⁃ 1α in breast cancer

作　　者：方玲玉 胡璟骅 文俊峰 韩诗琪 王雅丽 蒲卢兰[1,2,3] 李静佳 杨懿 邓世山 侯令密[2,3,4] 周方方[1,2,3] FANG Lingyu;HU Jinghua;WEN Junfeng;HAN Shiqi;WANG Yali;PU Lulan;LI Jingjia;YANG Yi;DENG Shishan;HOU Lingmi;ZHOU Fangfang(Institute of Basic Medicine and Forensic Medicine,North Sichuan Medical College,Nanchong 637100,China;Imaging Institute of North Sichuan Medical College,Nanchong 637100,China;Blogical Targeting Laboratory of Breast Cancer,Academician(Expert)Workstation,Afliated Hospital of North Sichuan Medical College,Nanchong 637100,China;不详)

机构地区：[1]川北医学院基础医学与法医学研究所,四川南充637100 [2]川北医学院影像研究所,四川南充637100 [3]川北医学院附属医院乳腺癌生物靶向研究室院士工作站,四川南充637100 [4]川北医学院附属医院甲乳外科,四川南充637100

出　　处：《实用医学杂志》2024年第16期2270-2276,共7页The Journal of Practical Medicine

基　　金：四川省科技创新苗子工程资助项目(编号:MZGC20230045);2022年医学影像四川省重点实验室开放课题(编号:MIKL202206);南充市科技局市校科技战略合作项目(编号:20SXQT0052);川北医学院2022年度四川省基层卫生事业发展研究中心资助项目(编号:SWFZ22-C-82);川北医学院2020年度校级科研发展计划项目(编号:CBY20-QA-Z13)。

摘　　要：目的探讨乳腺癌中泛素特异性蛋白酶20(ubiquitin-specific proteases 20,USP20)、低氧诱导因子1α(hypoxia inducible factor-1α,HIF-α)表达变化及意义。方法免疫组织化学及免疫荧光方法用于检测USP20与HIF-α阳性细胞的表达变化以及细胞定位,并分析它们之间的关系;shRNA-USP20慢病毒转染乳腺癌MDA-MB-231细胞构建乳腺癌MDA-MB-231 USP20过表达细胞系,siRNA-USP20构建乳腺癌MDA-MB-231USP20敲减细胞系,通过荧光定量PCR与Westem blot检测USP20基因及蛋白的过表达和敲减水平,并观察过表达以及敲减USP20前后HIF-α基因和蛋白的表达变化。结果USP20与HIF-α在乳腺癌组织中的阳性表达率分别为69.62%(55/79)和46.83%(37/79),而在邻近相对正常组织中的表达均为阴性,与邻近相对正常组织相比,乳腺癌组织中USP20与HIF-α阳性表达均显著增加,差异均有统计学意义(P<0.01),USP20与HIF-α阳性表达均主要在乳腺癌组织细胞质中,少量在细胞核;且乳腺组织中USP20表达与HIF-1α表达呈正相关(P<0.01);过表达USP20后HIF-α的mRNA的表达不变,蛋白表达显著增加(P<0.01)。敲减USP20后HIF-αmRNA表达轻微降低,但差异均无统计学意义(P>0.05),而蛋白表达水平显著降低(P<0.01)。结论乳腺癌中USP20和HIF-α表达均显著增加,且呈正相关关系,过表达USP20后HIF-α蛋白表达显著增加,敲减USP20后HIF-α蛋白表达显著降低,USP20可能通过影响HIF-α的表达进而促进乳腺癌发生发展。Objective To explore the changes and significane of USP20 and HIF-αexpression in breast cancer.Methods Following transfection of shRNA-USP20 lentivirus into breast cancer MDA-MB-231 cells,the gene and protein expression levels of USP20 were detected using fluorescence quantitative PCR and Western Blot.Subsequently,the overexpression of USP20 was observed to determine its effect on HIF-αexpression.Similarly,siRNA-USP20 was used to knock down USP20 in breast cancer MDA-MB-231 cells,followed by detection of gene and protein expression levels using fluorescence quantitative PCR and Western Blot.The subsequent changes in HIF-αexpression were then examined.Rusults The positive expression rates of USP20 and HIF-αin breast cancer tissues were 69.6%and 46.83%,respectively,while they were negatively expressed in the adjacent normal tissues,with statistically significant differences(P<0.01).The positive expressions of USP20 and HIF-αwere predomi⁃nantly observed in the cytoplasm of breast cancer tissue,with a smaller amount present in the nucleus.There was a significant positive correlation between USP20 and HIF-αin breast cancer.Following transfection of shRNA-USP20 lentivirus into MDA-MB-231 cells,both the protein and gene expression levels of USP20 significantly increased(P<0.01).Over-expression of USP20 did not affect HIF-αmRNA levels but led to a significant increase in HIF-αprotein expression(P<0.01).Conversely,siRNA-USP20 interference resulted in a significant decrease in both the protein and gene expression levels of USP20(P<0.01),without affecting HIF-αmRNA levels;however,it caused a notable reduction in HIF-αprotein expression(P<0.01).Conclusion The expression of USP20 exhib⁃ited a significant positive correlation with HIF-αin breast cancer.Overexpression of USP20 led to a substantial increase in HIF-αprotein expression,while knock-down of the USP20 gene resulted in a significant decrease in HIF-αprotein levels.Therefore,it can be inferred that USP20 may exert its influence on the development of br

关 键 词：乳腺癌 泛素特异性蛋白酶20 低氧诱导因子1Α 病理特征 

分 类 号：R737.9[医药卫生—肿瘤]