枯草溶菌素转化酶9在结直肠癌中的表达及其对细胞迁移的影响  

Investigation of proprotein convertase subtilisin/kexin type 9 expression in colorectal cancer and its impact on cell migration

作　　者：胡一佳 董海燕 向南霖 杨湘玲[1,2,3] 刘焕亮 HU Yijia;DONG Haiyan;XIANG Nanlin;YANG Xiangling;LIU Huanliang(Department of Clinical Laboratory,the Sixth Affiliated Hospital,Sun Yat-sen University,Guangzhou,Guangdong 510655,China;Guangdong Provincial Key Laboratory of Colorectal and Pelvic Floor Diseases,Guangdong Institute of Gastroenterology,the Sixth Affiliated Hospital,Sun Yat-sen University,Guangzhou,Guangdong 510655,China;Biomedical Innovation Center,the Sixth Affiliated Hospital,Sun Yat-sen University,Guangzhou,Guangdong 510799,CChhiinnaa)

机构地区：[1]中山大学附属第六医院检验科,广东广州510655 [2]中山大学附属第六医院广东省结直肠盆底疾病研究重点实验室,广东广州510655 [3]广州市黄埔区中六生物医学创新研究院,广东广州510799

出　　处：《热带医学杂志》2024年第5期635-640,I0002,共7页Journal of Tropical Medicine

基　　金：国家自然科学基金面上项目(82172337)。

摘　　要：目的探讨枯草溶菌素转化酶9(PCSK9)在结直肠癌(CRC)中的表达水平,及其对CRC细胞增殖、迁移的影响。方法通过分析癌症基因组图谱(TCGA)及高通量基因表达(GEO)数据库中PCSK9在正常组织和CRC组织中的表达差异;使用GenomicScape数据库分析PCSK9与CRC预后的关系;使用一个包含93例人类结肠癌和85例相邻正常组织的组织微阵列芯片,通过免疫组化的方法评估PCSK9的蛋白表达水平;使用TCGA数据库进行基因集富集分析(GSEA)、京都基因与基因组百科全书(KEGG)及基因本体(GO)富集分析;通过人重组蛋白刺激实验探索PCSK9在CRC中的作用。结果TCGA数据库分析结果显示,与正常组织相比,多种癌症如结肠腺癌、直肠腺癌的肿瘤组织中PCSK9的mRNA表达水平上调,差异均有统计学意义(P均<0.05)。4个CRC数据集(1个取自美国,1个取自德国,1个取自中国,1个取自挪威)的分析结果显示,与正常组织相比,PCSK9的mRNA水平在肿瘤组织中显著上调,差异均有统计学意义(t=12.07、12.72、12.13、13.38,P均<0.05)。与正常组织相比,CRC组织中PCSK9蛋白表达水平上调,差异有统计学意义(P<0.05)。与PCSK9高表达组比较,低表达组具有更高的总生存期,差异有统计学意义(HR=2.6,P<0.05)。GO及KEGG富集分析显示PCSK9相关基因富集在DNA复制及错配修复、细胞周期等多种生物过程中。Transwell迁移实验表明,20和100 ng/mL PCSK9预处理24 h后的CRC细胞HCT116迁移能力高于对照组(t=7.73、12.10),20和100 ng/mL PCSK9预处理24 h后的CRC细胞DLD1迁移能力高于对照组(t=3.79、6.51),差异均有统计学意义(P均<0.05)。细胞划痕实验表明,100 ng/mL PCSK9预处理24 h后的HCT116细胞愈合能力高于对照组(t=3.29),20和100 ng/mL PCSK9预处理24 h后的DLD1细胞愈合能力高于对照组(t=4.21、10.83),差异均有统计学意义(P均<0.05)。结论PCSK9在CRC组织中高表达,并与不良预后相关。重组PCSK9可能促进CRC细胞的迁移。Objective To explore the expression levels of proprotein convertase subtilisin/kexin type 9(PCSK9)in colorectal cancer(CRC)and its effects on the proliferation and migration of CRC cells.Methods This study analyzed the differential expression of PCSK9 in normal and CRC tissues by analyzing The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)databases;the relationship between PCSK9 expression and CRC prognosis was assessed using the Genomic Scape database;PCSK9 protein expression levels were evaluated using an immunohistochemical method in a tissue microarray chip containing 93 human colon cancer cases and 85 adjacent normal tissues.Gene set enrichment analysis(GSEA),Kyoto encyclopedia of genes and genomes(KEGG)and gene ontology(GO)enrichment analyses were conducted using the TCGA database;the role of PCSK9 in CRC was explored through human recombinant protein stimulation experiments.Results TCGA database analysis results showed that compared with the normal tissues,the m RNA expression levels of PCSK9 in various tumor types such as colon adenocarcinoma,rectum adenocarcinoma were up-regulated,the differences were statistically significant(all P<0.05).Analysis results of 4 datasets showed that compared with normal tissues,the m RNA expression levels of PCSK9 in tumor tissues of CRC patients from America(t=12.07),Germany(t=12.72),China(t=12.13)and Norway(t=13.38)were up-regulated,the differences were statistically significant(all P<0.05).The protein level of PCSK9 was significantly upregulated in tumor tissues of CRC than that in normal tissues(P<0.05).The low expression group had higher overall survival than the PCSK9 high expression group,the difference was statistically significant(HR=2.6,P<0.05).GO and KEGG enrichment analysis showed that PCSK9-related genes were enriched in various biological processes such as DNA replication,mismatch repair and cell cycle.Transwell migration assay indicated that the migration ability of CRC cells HCT116 treated with 20 and 100 ng/m L recombinant human PCSK9 were stron

关 键 词：枯草溶菌素转化酶9 结直肠癌 细胞迁移 

分 类 号：R735.3[医药卫生—肿瘤]