新抑癌基因GPD1在结肠癌细胞HCT-8中的作用及其预后潜力研究  

GPD1 represses colon cancer cell HCT-8 growth and indicates a favorable prognosis in colorectal cancer

作　　者：闵力 李恒存 郭庆东[1] 王星宇 张澍田[1] Min Li;Li Hengcun;Guo Qingdong;Wang Xingyu;Zhang Shutian(Department of Gastroenterology,Beijing Friendship Hospital,Capital Medical University,National Clinical Research Center for Digestive Disease,Beijing Digestive Disease Center,Beijing Key Laboratory for Precancerous Lesion of Digestive Disease,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院消化分中心国家消化系统疾病临床医学研究中心消化疾病癌前病变北京市重点实验室北京消化中心

出　　处：《首都医科大学学报》2019年第4期646-651,共6页Journal of Capital Medical University

基　　金：北京市医院管理中心消化内科学科协同发展中心重点项目(XXZ01)~~

摘　　要：目的探究新抑癌基因甘油-3-磷酸脱氢酶1(glycerol-3-phosphate dehydrogenase 1,GPD1)在结肠癌发生、发展中的作用及其与临床预后的关系。方法应用HCT-8结肠癌细胞系,以siRNA干扰GPD1表达后进行细胞功能实验,分别以MTS实验与集落形成实验检测细胞增生活力与集落形成能力,以划痕试验、Transwell实验探究GPD1对HCT-8细胞系迁移能力的影响。对TCGA数据库进行生物信息学分析,探究GPD1表达水平与结直肠癌患者临床预后的关系,并明确GPD1低表达与其编码基因甲基化之间的关系。结果干扰GPD1基因表达后,HCT-8细胞增生活力明显增强,重复实验差异具有统计学意义(P<0.01);集落形成实验结果显示,GPD1干扰后的HCT-8细胞集落数目增多且集落直径增大,差异具有统计学意义(P<0.01)。划痕实验与Transwell实验结果显示,敲低GPD1表达的HCT-8细胞迁移能力无显著变化,表明GPD1对结肠癌细胞迁移能力无影响。生物信息学分析结果显示,GPD1低表达的结直肠癌患者总存活率(χ^2=4.1,P=0.040)及无病存活率(χ^2=13.2,P<0.000 1)均明显低于GPD1高表达患者,且GPD1低表达水平是结直肠癌预后不良的独立风险因素(P<0.05)。甲基化分析结果显示,结直肠癌组织中GPD1低表达与其编码基因高甲基化呈负相关(P<0.05)。结论 GPD1在结肠癌中发挥抑制细胞增生与集落形成的作用,且GPD1低表达是结直肠癌患者不良预后的独立预测因素。Objective To explore the role of glycerol-3-phosphate dehydrogenase 1(GPD1) in the development of colorectal cancer and its relationship with clinical prognosis.Methods HCT-8 colon cancer cell line was used in this study.Cell function experiments were performed after GPD1 expression knockdown by using siRNA.Cell proliferation activity and colony formation ability were detected by MTS assay and colony formation assay respectively.The migration ability was determined applying wound scratch assay and transwell assay.The bioinformatics analysis of the TCGA database explored the relationship between the expression level of GPD1 and the clinical prognosis of colorectal cancer patients.The relationship between the low expression of GPD1 in colorectal cancer and the methylation level of its coding gene was also explored.Results After interference of GPD1 gene expression by siRNA,the proliferation ability of HCT-8 cells was significantly enhanced(P<0.01).The results of colony formation experiments showed that the colony number and size of HCT-8 cell colonies significantly increased after GPD1 siRNA treatment(P<0.01).The results of the wound scratch test and transwell assay showed that there was no significant change in the migration ability of GPD1 knockdown HCT-8 cells,indicating that GPD1 had no effect on the migration ability of colon cancer cells.Bioinformatics analysis showed that the overall survival(χ^2= 4.1,P=0.040) and diseasefree survival(χ^2= 13.2,P<0.000 1) of patients with low expression of GPD1 were significantly lower than those with high expression of GPD1.Also,low expression level of GPD1 was identified as an independent risk factor for poor prognosis of colorectal cancer(P <0.05).Methylation analysis showed that the low expression of GPD1 in colorectal cancer tissues was negatively correlated with hypermethylation of its coding gene(P<0.05).Conclusion GPD1 plays a role in inhibiting cell proliferation and colony formation in colon cancer,and low expression of GPD1 is an independent predictor of poor pr

关 键 词：结肠癌 GPD1 细胞增生 预后 

分 类 号：R735.35[医药卫生—肿瘤]