TBL1XR1介导上皮-间质转化和干细胞特性调控头颈部鳞状细胞癌细胞迁移和侵袭  

TBL1XR1 Regulates Migration and Invasion of Head and Neck Squamous Cell Carcinoma Cells via Inducing Epithelial-mesenchymal Transition and Stemness

作　　者：余长云[1] 薛彬彬 李金映 张晨[1] 张倩倩[1] 曹华[1] YU Chang-yun;XUE Bin-bin;LI Jin-ying;ZHANG Chen;ZHANG Qian-qian;CAO Hua(The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院,河南郑州450052

出　　处：《肿瘤学杂志》2022年第11期927-933,共7页Journal of Chinese Oncology

基　　金：国家自然科学基金青年基金项目(81402232);河南省重点研发与推广专项(科技攻关)(202102310116);河南省高等学校重点科研项目(20A320022);河南省医学科技攻关省部共建青年项目(SBGJ202103067)。

摘　　要：[目的]阐明转导素β1X连锁受体蛋白1 (transducin β-like 1 X-linked receptor 1,TBL1XR1)可介导上皮-间质转化和干细胞特性,从而调控头颈鳞癌细胞的迁移、侵袭。[方法]利用UALCAN数据库网站搜索TBL1XR1在头颈鳞癌组织及癌旁组织中的表达情况,应用Cbioprotal在线分析平台分析癌症基因组图集(The Cancer Genome Atlas,TCGA)数据库中523例头颈鳞癌RNA-SEQ数据。将慢病毒介导的TBL1XR1过表达载体、沉默载体和对应空白载体分别转染头颈鳞癌细胞Tu686。划痕愈合实验、Transwell迁移和侵袭实验评价Tu686细胞的迁移、侵袭能力;Western blot、qRT-PCR检测上皮-间质转化标志物表达;肿瘤球形成实验、qRT-PCR检测干细胞特性;抑制Wnt/β-catenin信号通路后检测TBL1XR1过表达引起的细胞迁移、侵袭、上皮-间质转化和干细胞特性。[结果] TBL1XR1 mRNA在头颈鳞癌组织中的表达明显高于相应癌旁组织(P<0.001)。过表达TBL1XR1的Tu686细胞划痕愈合能力(90%±4%vs 53%±6%)、Transwell迁移(0.68±0.04 vs 0.32±0.03)、侵袭能力(0.59±0.04 vs 0.26±0.04)较对照组显著性增强(P均<0.05)。TBL1XR1表达被抑制时,细胞划痕愈合能力(70%±4%vs 96%±5%)、Transwell迁移(0.32±0.06 vs 0.63±0.08)、侵袭能力(0.24±0.04 vs 0.52±0.05)较对照组显著性降低(P均<0.05)。TBL1XR1过表达后,Tu686细胞的间质细胞标志物Vimentin、N-cadherin表达升高,上皮细胞标志物E-cadherin表达降低;肿瘤球形成数较对照组明显增多(41±9 vs 13±4,P<0.05),肿瘤干细胞标志物ALDH1、CD44、CD133表达上调。抑制Wnt/β-catenin信号通路可部分逆转TBL1XR1过表达引起的迁移、侵袭、上皮-间质转化及干细胞特性改变。[结论] TBL1XR1可在体外介导上皮-间质转化和干细胞特性,进而促进头颈鳞癌细胞迁移、侵袭,其调控机制可能与Wnt/β-catenin信号通路相关。[Objective] To investigate the regulatory effect of transducin β-like 1 X-linked receptor 1(TBL1XR1) on the migration and invasion of head and neck squamous cell carcinoma(HNSCC) cell and its mechanism. [Methods] The TBL1XR1 gene expression level in HNSCC and paracancerous tissues was obtained from UALCAN database. Data of 523 patients with HNSCC in the TCGA database were analyzed by cbioprotal online analysis platform. The relationship between the TBL1XR1 genetic alteration and lymph node stage was analyzed. HNSCC Tu686 cells were transfected with lentivirus mediated TBL1XR1 overexpression vector, downregulation vector and control vector. The wound healing assay, transwell migration and invasion assays were employed to measure the effects of TBL1XR1 on the migratory and invasive abilities of Tu686cells. Western blot, qRT-PCR were used to investigate the effects of TBL1XR1 on epithelial-mesenchymal transition makers. The tumor sphere formation assay and qRT-PCR assay were performed to investigate the stemness of Tu686 cells. The inhibitor was used to block the activation of Wnt/β-catenin signaling pathway in Tu686 cells, then the changes of migration and invasion abilities, epithelial-mesenchymal transition, stemness were measured. [Results] The expression of TBL1XR1 in HNSCC was higher than those in paracancerous tissues(P<0.001). The abilities of wound healing(90%±4% vs 53%±6%), transwell migration(0.68±0.04 vs0.32±0.03) and invasion(0.59±0.04 vs 0.26±0.04) of TBL1XR1-overexpression Tu686 cells were significantly enhanced compared with control cells(all P<0.05). On the contrary, significantly decreased wound healing(70%±4% vs 96%±5%), migration(0.32±0.06 vs 0.63±0.08) and invasive abilities(0.24±0.04 vs 0.52±0.05)were observed when TBL1XR1 expression was inhibited( all P<0.05). After TBL1XR1 was up-regulated, the expression of mesenchymal markers Vimentin, N-cadherin were elevated, the expression of epithelial maker E-cadherin was reduced, the number of tumor sphere formation(41±9 vs 13±4, P<0.0

关 键 词：头颈部肿瘤 癌 鳞状细胞 转导素β1X连锁受体蛋白1 上皮-间质转化 肿瘤干细胞 WNT/Β-CATENIN 

分 类 号：R739.6[医药卫生—肿瘤]