含有WW结构域的氧化还原酶的基因在骨肉瘤组织中表达的临床意义及其对骨肉瘤的调控作用  

Expression of WW domain-containing oxidoreductase in osteosarcoma tissues and its effect on proliferation and metastasis of osteosarcoma cells

作　　者：许峰[1] 闻嘉[2] 谢祎 李家平[4] 李甲振[2] 张岩[2] 卢新昌[2] 张翼[2] 刘永奎[2] 王帝 李哲[2] 元耀博 李隆卿 张卫红[2] Xu Feng;Wen Jia;Xie Yi;Li Jiaping;Li Jiazhen;Zhang Yan;Lu Xinchang;Zhang Yi;Liu Yongkui;Wang Di;Li Zhe;Yuan Yaobo;Li Longqing;Zhang Weihong(Department of Neurosurgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Orthopedics,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Neurology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Interventional Oncology,the First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510030,China)

机构地区：[1]郑州大学第一附属医院神经外科,450052 [2]郑州大学第一附属医院骨科,450052 [3]郑州大学第一附属医院神经内科,450052 [4]中山大学附属第一医院肿瘤介入科,广州510030

出　　处：《中华实验外科杂志》2021年第4期745-751,共7页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划联合共建项目(2018020083)。

摘　　要：目的探讨含有WW结构域的氧化还原酶的基因(WWOX)在骨肉瘤组织中的表达与患者临床病理特征的相关性,研究WWOX在小鼠中对人骨肉瘤生长和转移的影响及其分子机制。方法收集2011年1月至2018年12月来自郑州大学第一附属医院的骨肉瘤患者112例,采用免疫组织化学和反转录-聚合酶链反应(RT-PCR)检测骨肉瘤组织中WWOX的表达,并分析WWOX表达与骨肉瘤临床病理特征之间的关系,然后建立人骨肉瘤免疫缺陷鼠荷瘤模型,分别注射空白载体、慢病毒载体的WWOX和慢病毒载体的WWOX短发卡RNA(shRNA),比较3组免疫缺陷鼠肿瘤体积变化,荷瘤标本用qRT-PCR和蛋白质印迹法(Western blot)评估WWOX在小鼠原位瘤和肺转移瘤中的表达情况,CT分析肺转移情况。Western blot检测WWOX敲低组与对照组比较相关蛋白的表达量变化。对计量资料符合正态分布的使用独立样本t检验进行差异分析,不符合正态分布的使用非参数检验进行差异分析,等级资料使用非参数检验进行差异分析,计数资料使用检验进行差异分析。不同组之间的差异采用one-way ANOVA方法分析。结果WWOX的表达与患者年龄、性别和发病部位等临床病理特征无关,而WWOX高表达与较低的微血管密度(MVD)(P<0.05)和较小的肿瘤体积明显相关(P<0.05)。此外,WWOX高表达患者的总生存期(OS)和无病生存期(DFS)均好于WWOX低表达患者(P值均<0.05),差异有统计学意义。瘤内注射WWOX的小鼠原位肿瘤WWOX表达与对照组和GDPAH的比值分别为1.43±0.17和2.31±0.26,表达明显高于对照组,MVD分别为19.34±3.71与11.71±1.72,表达明显低于对照组,肿瘤生长缓慢,肺转移瘤肿瘤个数、WWOX表达和MVD与对照组差异无统计学意义,瘤内注射WWOX shRNA的小鼠原位肿瘤WWOX表达和GDPAH的比值为0.76±0.19,表达明显低于对照组,MVD为28.50±3.44,表达明显高于对照组,肿瘤生长加快,肺转移瘤数量为13.00±4.19,表达明显�Objective To study the correlation between WW domain-containing oxidoreductase(WWOX)expression and clinicopathological features of osteosarcoma,and explore the effect of WWOX on the proliferation and metastasis of human osteosarcoma in mice.Methods The expression of WWOX in osteosarcoma tissues was detected by reverse transcriptase-polymerase chain reaction(RT-PCR)and immunohistochemistry.The correlation between the WWOX expression and clinicopathological features of osteosarcoma was evaluated.The osteosarcoma cells 143B were injected into the tibias of immunodeficient mice to establish the osteosarcoma xengrafted model.Lentivirus,lentivirus-delivered WWOX and lentivirus-delivered WWOX short hairpin RNA(shRNA)were injected into tumor respectively.The tumor growth was recorded and compared.The number of pulmonary nodules was assessed using micro CT.For the measurement data that conform to the normal distribution,the independent sample t test was used for difference analysis;for the measurement data that do not conform to the normal distribution,the nonparametric test was used for difference analysis;for the grade data,the nonparametric test was used for difference analysis;for the count data,the test was used for difference analysis.The differences between different groups were analyzed by one-way ANOVA.Results There was no significant correlation between WWOX expression and some clinicopathological features,including gender,age,primary site,etc.The high WWOX expression was correlated with low microvascular density(MVD)and small tumor volume.The overall survival(OS)and disease-free survival in patients with high WWOX expression was obviously longer than those of patients with low WWOX expression.The growth rate of primary tumor in immunodefient mice was significantly lower in WWOX group and significantly higher in WWOX shRNA group than in vector group.The number of pulmonary nodule was significantly greater in WWOX shRNA group than in vector group.However,there was no significant difference in the number of pulmon

关 键 词：骨肉瘤 预后 肺转移 细胞外信号调节激酶信号通路 

分 类 号：R738.1[医药卫生—肿瘤]