TDO2基因对鼻咽癌细胞增殖、迁移和凋亡能力的影响及其机制  

Effects and mechanism of TDO2 gene on proliferation,migration and apoptosis of nasopharyngeal carcinoma cells

作　　者：刘翔 罗燕贞 周兰柱[1] 孙哲[1] 吴俊 王文忠[1] LIU Xiang;LUO Yanzhen;ZHOU Lanzhu;SUN Zhe;WU Jun;WANG Wenzhong(Department of Otolaryngology Head and Neck Surgery,First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院第一附属医院耳鼻咽喉头颈外科,蚌埠233000

出　　处：《山西医科大学学报》2023年第1期53-59,共7页Journal of Shanxi Medical University

基　　金：安徽省高等学校自然科学研究项目(KJ2020A0568);蚌埠医学院研究生科研创新项目(Byycx21104)。

摘　　要：目的探讨色氨酸-2,3-双加氧酶2(tryptophan-2,3-dioxygenase2,TDO2)对鼻咽癌细胞系CNE-2Z和HNE-1细胞生物学行为的影响及其作用机制。方法通过TIMER2数据库分析TDO2基因在不同癌症或特定癌症亚型中的表达状态。培养人正常鼻咽黏膜上皮细胞系NP69、鼻咽癌细胞系5-8F、CNE-2Z和HNE-1。实验中对CNE2Z和HNE-1细胞分别转染TDO2干扰序列和阴性对照序列,分别命名为si-TDO2组和NC组。采用实时荧光定量PCR(qRT-PCR)方法检测TDO2的mRNA相对表达水平;采用细胞计数试剂盒(CCK-8)检测细胞存活率,细胞集落克隆实验检测集落数量;Transwell法和细胞划痕实验检测细胞迁移能力;蛋白质印迹实验(Western blot)检测鼻咽癌细胞中上皮间充质化(EMT)相关蛋白E-cadherin、N-cadherin及Vimentin及凋亡相关蛋白Bax、Bcl-2相对表达量。结果与正常鼻咽黏膜上皮细胞NP69相比,在CNE-2Z和HNE-1细胞中TDO2的mRNA相对表达量明显上调(P<0.001)。与NC组比较,si-TDO2组细胞存活率降低,克隆细胞集落明显减少(P<0.05);与NC组相比,si-TDO2组细胞迁移数量较少,迁移指数较小(P<0.05);与NC组相比,si-TDO2组EMT相关蛋白中N-cadherin和Vimentin蛋白相对表达水平降低,E-cadherin蛋白相对表达水平升高,凋亡相关蛋白Bax蛋白相对表达量升高,Bcl-2蛋白的相对表达量降低(P<0.05)。结论TDO2可促进鼻咽癌细胞株CNE-2Z和HNE-1细胞增殖、迁移能力,并抑制细胞凋亡,其机制可能通过促进EMT进行调控。Objective To investigate the effect of tryptophan-2,3-dioxygenase 2(TDO2)on the biological behaviors of CNE-2Z and HNE-1 cells in nasopharyngeal carcinoma cell lines and its mechanism.Methods The expression of TDO2 gene in different cancers or specific cancer subtypes was analyzed based on TIMER2 database.In the experiment,the normal nasopharyngeal mucosal epithelial cell line NP69,the nasopharyngeal carcinoma cell lines 5-8F,CNE-2Z and HNE-1 were cultured.CNE-2Z and HNE-1 were transfected with TDO2 interference sequence and negative control sequence in si-TDO2 group and NC group,respectively.Real-time PCR(qRT-PCR)method was used to detect the relative expression level of TDO2 mRNA.The cell counting kit(CCK-8)was used to detect the cell viability,and colony cloning assays were used to measure the colony number.Transwell method and cell scratch experiment were performed to detect the cell migration.Western blot was used to measure the relative expression of EMT-associated proteins E-cadherin,N-cadherin and Vimentin and apoptosis-related proteins Bax and Bcl-2 in nasopharyngeal carcinoma cells.Results TDO2 mRNA expression in CNE-2Z and HNE-1 cells was significantly higher than that in normal nasopharyngeal mucosa epithelial cell NP69(P<0.001).Compared with NC group,the cell survival rate and the colony of cloned cells were significantly decreased in si-TDO2 group(P<0.05).Compared with NC group,the cell number of migration and the migration index were decreased in si-TDO2 group(P<0.05).Compared with NC group,the relative expression levels of N-cadherin and Vimentin were reduced in si-TDO2 group,while the relative expression level of E-cadherin protein was increased(P<0.05),and the relative expression of Bax protein was increased,while the relative expression of Bcl-2 protein was reduced(P<0.05).Conclusion TDO2 can promote the proliferation and migration abilities of nasopharyngeal carcinoma cell lines CNE-2Z and HNE-1 cells,and inhibit the apoptosis,which may be regulated by promoting EMT.

关 键 词：鼻咽癌 TDO2 增殖 迁移 凋亡 

分 类 号：R739.6[医药卫生—肿瘤]