YTH结构域家族蛋白2在肝癌组织的表达及其临床意义  

Expression andclinical significance of YTH domain containing family 2 in hepatocellular carcinoma

作　　者：马志立[1] 刘权焰[1] 陈白杨 刘朋朋[1] 古阳 马栋 刘志苏[1] Ma Zhili;Liu Quanyan;Chen Baiyang;Liu Pengpeng;Gu Yang;Ma Dong;Liu Zhisu(Department of General Surgery, Zhongnan Hospital, Wuhan University, Wuhan 430071, China)

机构地区：[1]武汉大学中南医院普外科,430071

出　　处：《中华实验外科杂志》2019年第2期319-322,共4页Chinese Journal of Experimental Surgery

摘　　要：目的 观察YTH结构域家族蛋白2(YTHDF2)在肝癌组织的表达和其患者预后的关系及其对肝癌细胞增殖和侵袭迁移的影响。方法 采用生物信息学方法和实时荧光定量聚合酶链反应(RT-qPCR)以及免疫组织化学检测配对肝癌组织及癌旁组织中YTHDF2的表达,分析其与患者临床特征的关系。敲除肝癌细胞中的YTHDF2,使用细胞计数试剂盒(CCK-8)实验检测细胞增殖能力,Transwell和细胞划痕实验检测细胞的侵袭转移能力,使用Western blot分析上皮-间充质转化(EMT)相关蛋白,E-钙黏蛋白(E-cadherin)和N-钙黏蛋白(N-cadherin)的表达量变化。结果 生物信息学分析表明YTHDF2在肝癌组织中高表达且与患者预后相关。YTHDF2在肝癌组织中的表达水平较癌旁组织高(18.23±3.51比13.21±2.73,P<0.01),其表达水平与肿瘤的巴塞罗那分期,TNM分期和有无门静脉癌栓(PVTT)相关(P均<0.01)。CCK-8实验显示敲除YTHDF2后24 h测得波长450 nm处吸光度值较对照组降低,HepG2中测得为(1.129±0.178比1.571±0.226,P<0.01),Huh7为(1.227±0.251比1.815±0.302,P<0.05)。Transwell实验显示敲除YTHDF2后穿过小室的细胞数减少,细胞划痕实验显示敲除YTHDF2后细胞划痕愈合度降低。敲除YTHDF2后,E-cadherin相对表达量较对照组显著升高(1.780±0.319比0.916±0.173,P<0.05),而N-cadherin的相对表达量明显降低(0.821±0.217比4.697±0.398,P<0.01)。结论 YTHDF2在肝癌中高表达,可促进肝癌的发生发展以及增强肝癌细胞的增殖和侵袭迁移能力。Objective To analyze the expression of YTH domain containing family 2 (YTHDF2) in the hepatocellular carcinom (HCC) and the relationship between the expression of YTHDF2 and prognosis. To detect the influence of YTHDF2 in cell proliferation, migration and invasion.Methods We detected the expression of YTHDF2 among the paired HCC tissue and adjacent tumor tissue with bioinformatics, real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) and immunohistochemistry. To analyze the relationship between YTHDF2 expression and clinical characteristics. We established the HCC cells with YTHDF2 knockdown and analyzed the cell proliferation with cell counting kit-8 (CCK-8) assay and detect the cell migration with Transwell and wound scratch assay. To analyze the expression of E-cadherin and N-cadherin that associated with epithelial-mesenchymal transition (EMT) with western blotting.Results Bioinformatics analysis showed the high expression of YTHDF2 in HCC tumor and its relationship with poor prognosis. We found that the HCC tumor tissue has a higher expression of YTHDF2 in compared with non-tumor adjacent tissue (17.76±4.78 vs. 12.10±3.43, P<0.01). The expression of YTHDF2 is positively related with BCLC stage, TNM stage and portal vein tumor thrombosis (PVTT) (P<0.01). CCK-8 assay demonstrated that the absorbance at 450 nm of the cell with YTHDF2 knockdown is lower than the negative control, while HepG2 is (1.129±0.178 vs. 1.571±0.226, P<0.01) and Huh7 (1.227±0.251 vs. 1.815±0.302, P<0.05). Transwell assay showed that the number of cell that diffuses through the membrane of cell with YTHDF2 knockdown is lower than the negative control. The wound scratch assay showed that the percent of wound area in cell with YTHDF2 knockdown is lower. We detected the HCC cells with YTHDF2 konckdown and found it had a higher relative expression of E-cadherin (1.780±0.319 vs. 0.916±0.173, P<0.05) and a lower expression of N-cadherin (0.821±0.217 vs. 4.697±0.398, P<0.01).Conclusion The high expression of

关 键 词：YTH结构域家族蛋白2 肝癌 上皮-间充质转化 

分 类 号：R735.7[医药卫生—肿瘤]