摘要
目的旨在探讨乳腺癌中FAS相关因子家族成员2(FAF2)基因表达与患者预后关联性以及has-miR-9-5p对FAF2基因表达的直接调控作用。方法基于肿瘤基因组图谱(TCGA)和基因表达综合数据库(GEO)数据分析并验证FAF2基因高表达与乳腺癌患者预后关联性;高表达has-miR-9-5p后,RT-qPCR和蛋白质印迹法检测FAF2表达变化;通过双荧光素酶报告系统验证has-miR-9-5p对FAF2的靶向抑制作用。利用GraphPad 7.0对数据进行统计学分析。结果 DNA芯片数据分析显示FAF2基因高表达与乳腺癌患者预后不良有关联(HR=1.44,95%CI为1.31~1.59,P<0.001),RNA测序数据分析也显示FAF2基因高表达与乳腺癌患者预后不良有关联(HR=1.40,95%CI为1.13~1.75,P=0.002);细胞转染实验中,has-miR-9-5p高表达组FAF2mRNA水平(0.25±0.08)较对照组(1.07±0.04)下降,差异有统计学意义,t=16.02,P<0.001,高表达组蛋白水平(0.19±0.02)较对照组(0.32±0.06)亦下降,差异有统计学意义,t=3.83,P=0.019。双荧光素酶报告实验中,转染FAF2野生型WT-pmiRGLO质粒后,hsa-miR-9-5p高表达组荧光相对强度(0.32±0.09)较对照组(1.22±0.09)降低,差异有统计学意义,t=12.03,P<0.001;转染FAF2突变型MT-pmiRGLO质粒后,hsa-miR-9-5p高表达组荧光相对强度(1.25±0.05)与对照组(1.14±0.08)比较,差异无统计学意义,t=0.76,P=0.489。结论乳腺癌中has-miR-9-5p可靶向抑制潜在预后不良标志物FAF2基因的表达。
Objective To reveal the correlation between gene expression of FAS related factor family member 2(FAF2)and prognosis of breast cancer patients,and effect of has-miR-9-5 p on the expression of FAF2.Methods The TCGA and GEO data were used to analyze and verify the correlation between high expression of FAF2 and prognosis of breast cancer patients.FAF2 expression was detected by RT-qPCR and Western blot after high expression of has-miR-9-5 p.Finally,the dual-luciferase reporter system was used to verify the direct inhibition of has-miR-9-5 p on FAF2.GraphPad 7.0 was used for statistical analysis of the data.Results DNA chip data analysis showed that the high expression of FAF2 gene was associated with poor prognosis of breast cancer patients(HR=1.44,95%CI:1.31-1.59,P<0.001),and RNA sequencing data analysis also showed that high FAF2 gene expression was associated with the poor prognosis of breast cancer patients(HR=1.40,95%CI:1.13-1.75,P=0.002);In the cell transfection experiment,the FAF2 mRNA level(0.25±0.08)of the has-miR-9-5 p high expression group was significantly lower than that of the control group(1.07±0.04,t=16.02,P<0.001),the protein level(0.19±0.02)also decreased significantly(0.32±0.06,t=3.83,P=0.019).In the dual luciferase reporter experiments,after transfection of FAF2 wild-type WT-pmiRGLO plasmid,the relative fluorescence intensity of hsa-miR-9-5 p high expression group(0.32±0.09)was significantly lower than that of the control group(1.22±0.09),t=12.03,P<0.001;after transfection of FAF2 mutant MT-pmiRGLO plasmid,the relative fluorescence intensity of hsa-miR-9-5 p high expression group(1.25±0.05)was not significantly different from the control group(1.14±0.08),t=0.76,P=0.489.Conclusion In breast cancer,has-miR-9-5 p directly inhibits FAF2 expression which is a potential marker of poor prognosis.
作者
张虎
夏伟
陈根林
凌存保
陈文艳
朱金玲
张淑红
ZHANG Hu;XIA Wei;CHEN Gen-lin;LING Cun-bao;CHEN Wen-yan;ZHU Jin-ling;ZHANG Shu-hong(Department of Basic Medical,Jiangsu Vocational College of Medicine,Yancheng 224005,China;Department of Biology,Basic Medical College,Jiamusi University,J iamusi 154007,China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2021年第15期1138-1143,共6页
Chinese Journal of Cancer Prevention and Treatment
基金
江苏省高等学校自然科学研究面上项目(18KJB310002)
黑龙江省卫生计生委科研课题(2018509,2017406)
盐城市科学科技发展计划(YK2019047,YK2020063)
江苏医药职业学院校级自然科学基金(20204101)
江苏省高等职业教育高水平专业群建设项目[苏教职函(2021)1号]。
