摘要
目的:探讨m6A甲基化修饰结合蛋白YTHDF2(YTH domain-containing family protein 2)在肝癌(Hepatocellular carcinoma,HCC)组织中的表达及临床意义。方法:1)采用Western blot和实时荧光定量PCR(Quantitative Real Time PCR, qRT-PCR)检测20对肝癌和癌旁组织中YTHDF2在蛋白和m RNA水平的表达情况。2)通过免疫组织化学(Immunochemistry, IHC)检测40对肝癌和癌旁组织芯片YTHDF2的表达情况,并以H-score评分法进行半定量分析。3)通过基因表达谱数据动态分析数据库GEPIA(http://gepia.cancer-pku.cn/)分析YTHDF2在肝癌组织中的表达及对患者生存预后的影响;通过肿瘤免疫评估资源数据库TIMER(https://cistrome.shinyapps.io/timer/)分析YTHDF2与肝癌免疫微环境的相关性。结果:相比于癌旁肝组织,YTHDF2在肝癌组织中蛋白和m RNA水平均显著高表达(P<0.05);GEPIA数据库分析验证:YTHDF2肝癌组织表达水平高于正常肝组织,且YTHDF2在肝癌不同分期中表达水平均高于正常肝组织(P=0.0206),Stage Ⅲ期最为明显;YTHDF2高表达患者的总体生存率(Overall Survival,OS)和无复发生存率(Disease Free Survival, DFS)均较差(P=0.00027和P=0.013);TIMER数据库分析表明:YTHDF2在肝癌免疫微环境中与各类免疫细胞呈正相关(P<0.05),但肝癌患者的累积存活率不受免疫细胞的影响(P>0.05),YTHDF2高表达对患者的生存预后具有不良影响(P=0.007)。结论:YTHDF2在肝癌组织中显著高表达,且YTHDF2高表达水平对肝癌患者生存预后具有不良影响,YTHDF2可作为肝癌临床预后判断的分子标志物,为今后肝癌治疗靶点提供新的策略。
Objective:To investigate the expression and clinical significance of m6A methylated modification binding protein YTHDF2(YTH domain-containing family protein 2)in hepatocellular carcinoma(HCC).Methods:1)The protein and mRNA expression of YTHDF2 in 20 pairs of HCC tissue and para-cancer tissue was detected by western blot and quantitative real-time PCR(qRT-PCR).2)The expression of YTHDF2 in microarray with 40 pairs of HCC tissue and para-tumor tissue was detected by immunochemistry.H-score was used for semi-quantitative analysis.3)The expression of YTHDF2 in HCC as well as its influence on survival prognosis of patients were analyzed by the GEPIA database(http://gepia.cancer-pku.cn/);The correlation between YTHDF2 and tumor immune microenviron-ment was analyzed by the TIMER database(https://cistrome.shinyapps.io/timer/).Results:The protein and mRNA expression of YTHDF2 in HCC was significantly higher than that in para-tumor tissue(P<0.05).GEPIA analysis verified that the expression of YTHDF2 in HCC was higher than that in normal liver tissue;the expression of YTHDF2 in different stages of HCC was also higher than that in normal liver tissue(P=0.0206),which was most obvious at Stage III.The overall survival(OS)and disease-free survival(DFS)of patients with high YTHDF2 expression were poor(P=0.00027 and P=0.013).TIMER database analysis showed that YTHDF2 was positively correlated with various types of immune cells in the immune microenvironment of HCC(P<0.05),but cumulative survival of HCC patients was not affected by immune cells(P>0.05),and the overexpression of YTHDF2 had a negative influence on the survival progno-sis of patients(P=0.007).Conclusions:YTHDF2 was obviously overexpressed in HCC tissue.The overexpression of YTHDF2 had an ad-verse effect on the survival prognosis of HCC patients,suggesting that YTHDF2 may be a predictive molecular marker for clinical prog-nosis of HCC and providing a new strategy for therapeutic targets of HCC in the future.
作者
王月帆
葛春梅
尹昊瓒
戴智慧
董峻鹏
季曼
李雯
杨富
WANG Yue-fan;GE Chun-mei;YIN Hao-zan;DAI Zhi-hui;DONG Jun-peng;JI Man;LI Wen;YANG Fu(The Department of Medical Genetics,Naval Medical University,Shanghai,200433,China;The Third Department of Hepatic Surgery,Eastern Hepatobiliary Surgery Hospital,Shanghai,200438,China)
出处
《现代生物医学进展》
CAS
2021年第9期1601-1606,共6页
Progress in Modern Biomedicine
基金
国家重点研发计划项目(2016YFC1302303)
国家自然科学基金项目(81972657,81672345)。
