PR结构域在PRDM16抑制结直肠癌迁移中的作用研究  

Effect of PR domain on the anticancer activity of PRDM16 in colorectal cancer cells

作　　者：杨锦燮 张隽[3] 谌倩芸 杨芝春 向跃芸[4] YANG Jin-xie;ZHANG Jun;CHEN Qian-yun;YANG Zhi-chun;XIANG Yue-yun(Department of Pharmacology,Xiangya School of Pharmaceutical Sciences,Central South University,Changsha 410078;Hunan Key Laboratory of Cardiovascular Research,Changsha 410078;Department of Oncology,the Third Xiangya Hospital,Central South University,Changsha 410013;Department of Clinical Laboratory,Hunan Maternal and Child Healthcare Hospital,Changsha 410003)

机构地区：[1]中南大学湘雅药学院药理学系,长沙410078 [2]湖南省心血管研究重点实验室,长沙410078 [3]中南大学湘雅三医院肿瘤科,长沙410013 [4]湖南省妇幼保健院检验科,长沙410003

出　　处：《中南药学》2024年第11期2837-2845,共9页Central South Pharmacy

基　　金：国家自然科学基金(No.81673491);湖南省自然科学基金(No.2022JJ30792);湖南省妇幼保健院锐新培育计划(No.2021RX14)。

摘　　要：目的探究PRDM16及其PR结构域在结直肠癌增殖迁移中的作用。方法应用TCGA数据库和生物信息学分析总PRDM16(tPRDM16)与结直肠癌的相关性;免疫组化及PCR检测结直肠癌组织中PRDM16表达;Westernblot检测结直肠癌细胞株中PRDM16本底表达水平,然后在高表达株敲低结肠癌细胞株中总tPRDM16,在低表达株中分别过表达全长PRDM16(fPRDM16)与短型PRDM16(sPRDM16),然后通过CCK8、平板克隆实验以及划痕实验、Transwell迁移实验检测fPRDM16与sPRDM16对结直肠癌细胞增殖和迁移能力的影响。结果生信分析结果显示,总tPRDM16水平(包括fPRDM16、sPRDM16)是结直肠癌的危险因素,也是免疫检查点抑制剂疗效的预测因素。临床研究结果显示,与正常组织相比,tPRDM16、fPRDM16和UCP1在结直肠癌组织中高表达,且UCP1mRNA水平与fPRDM16、sPRDM16mRNA均成正相关。敲低SW480细胞中PRDM16,发现tPRDM16与fPRDM16皆下调,伴细胞迁移能力下降,但对细胞增殖并无明显影响。在HCT116细胞中进行fPRDM16过表达则可抑制肿瘤细胞迁移,伴线粒体解偶联蛋白UCP1的蛋白表达水平升高;而过表达sPRDM16则可降低UCP1蛋白水平且促进细胞迁移。进一步研究发现,敲低UCP1可促进HCT116细胞的迁移。结论PR结构域是PRDM16抑制结直肠癌迁移的功能结构域,其机制可能与维持UCP1蛋白的正常表达水平有关。Objective To determine the effect of PRDM16 on the proliferation and migaration of colorectal cancer(CRC)cells and the role of PR domain in the anti-migaration effect of PRDM16.Methods The relationship between total PRDM16(tPRDM16,including two isoforms fPRDM16 and sPRDM16)and CRC was analyzed with TCGA database and bioinformatics.The expression of PRDM16 protein was detected by immunohistochemistry or PRC Western blot,and then was knocked down in SW480 cells,and over-expressed full-length PRDM16(fPRDM16)or short PRDM16(sPRDM16)in HCT116 cells respectively.CCK8,wound healing assay,colony formation assay and transwell migration assay were used to determine the proliferation and migration of CRC cells.Results Bio-informatics analysis showed that tPRDM16 level might be considered as both a risk factor for CRC and a predictive factor for treatment with immune checkpoint inhibitors.Clinical studies showed that tPRDM16,fPRDM16 and UCP1 expressions were all high in CRC tissues,and UCP1 mRNA was positively correlated with both tPRDM16 and fPRDM16.In vitro experiments showed that down-regulation of tPRDM16 via shRNA interference inhibited the migration of SW480 cells but didnot affect the proliferation.It was observed that over-expression of fPRDM16 inhibited the migration of HCT116 cells,while over-expression of short PRDM16(sPRDM16)promoted the migration.Over-expression of fPRDM16 also up-regulated UCP1 expression,while over-expression of sPRDM16 decreased UCP1 level in HCT116 cells.Moreover,the knockdown of UCP1 promoted the migration of HCT116 cells.Conclusion PR domain is the functional domain of PRDM16 protein in inhibiting the migration of colorectal cancer,whose mechanism may be related to maintaining the normal expression of UCP1 protein.

关 键 词：结直肠癌 PRDM16 PR结构域 UCP1 

分 类 号：R96[医药卫生—药理学] R735.34[医药卫生—药学]