标题：OAZ1基因稳定表达SCC15细胞株的构建及其对舌鳞状细胞癌上皮间质化的影响

      作者：王星 1，姜立 2，张沄 1，陈肖燕 2    (1.西南医科大学附属医院炎症与变态反应实验室，四川 泸州 646000；2.广东省人民医院医学研究中心，广东 广州 510080)

      卷次： 2017年28卷13期

      【摘要】 目的 探索鸟氨酸脱羧酶抗酶(OAZ1)对舌鳞癌细胞株 SCC15上皮间质化的影响及相关机制。方法 构建框移位点突变的OAZ1过表达慢病毒表达载体 pLVX-Neo-OAZ1-IRES-ZsGreen，包装病毒并感染SCC15细胞，采用有限稀释法挑取荧光强度较高的单克隆细胞株；利用RT-PCR和Western blot检测OAZ1以及上皮间质化相关基因(E-cadherin、Vimentin、N-cadherin、Zeb1、Snail、Twist1、TGF-β1、Smad1)的mRNA水平和蛋白水平。结果 成功构建了OAZ1基因稳定表达的SCC15细胞株，且OAZ1的过表达可以抑制Vimentin、N-cadherin、TGF-β1的mRNA和蛋白水平表达；此外，OAZ1可以促进Smad1的蛋白水平降低。结论 OAZ1可以抑制舌鳞癌细胞的上皮间质化，其机制可能涉及TGF-β1信号通路和Smad1信号通路的抑制。
      【关键词】 鸟氨酸脱羧酶抗酶；舌磷状细胞癌；慢病毒；上皮间质化
      【中图分类号】 R739.86 【文献标识码】 A 【文章编号】 1003—6350（2017）13—2065—07

Establishment of SCC15 cell line stably expressing human OAZ1 gene and its influence on the epithelialmesenchymal transition of tongue squamous cell carcinoma.
WANG Xing 1, JIANG Li 2, ZHANG Yun 1, CHENXiao-yan 2. 1. Laboratory of Inflammation and Allergy, the Affiliated Hospital of Southwest Medical University, Luzhou646000, Sichuan, CHINA; 2. Medical Research Center, Guangdong General Hospital, Guangzhou 510080, Guangdong,CHINA
【Abstract】 Objective To investigate the role of OAZ1 in epithelial mesenchymal transition (EMT) in tonguesquamous cell carcinoma cell lines SCC15 and explore its related mechanism. Methods The lentivirus vectorpLVX-Neo-OAZ1-IRES-ZsGreen in which the target gene OAZ1 was modified at the frameshift site was constructed,packaged lentivirus and infected SCC15 cells, and the monoclonal cell lines with high fluorescence intensity were ob-tained by limited diluted method. Real-time PCR and Western blot was used to detect the expression of OAZ1 and thegenes (E-cadherin, Vimentin, N-cadherin, Zeb1, Snail, Twist1, TGF-β1, Smad1) related with EMT. Results The SCC15cell line with stable expression of OAZ1 gene was successfully constructed. The overexpression of OAZ1 inhibited theprotein level and mRNA level of the Vimentin, N-cadherin, TGF-β1. Furthermore, the OAZ1 would promote the degrada-tion of Smad1. Conclusion OAZ1 can inhibit the EMT of tongue squamous cell carcinoma cell lines through the possi-ble mechanism related with the inhibition of TGF-β1 and Smad1 pathway.
      【Key words】 Ornithine decarboxylase antizyme 1 (OAZ1); Tongue squamous cell carcinoma (TSCC); Lentivi-rus; Epithelial mesenchymal transition (EMT)·论 著·doi:10.3969/j.issn.1003-6350.2017.13.001基金项目：国家自然科学基金(编号：30901757)