Liu Fei, Tang Fengzhu, Lan Jiao, Jiao Wei, Si Yongfeng, Lu Wensheng, Mo Mingzheng, Li Bing, Lu Jinlong, Wei Jiazhang, Qin Ying, Xiao Ruiping, Zhang Benjian, Wang Yongli, Xiong Weiming
1 Research Center of Medical Sciences, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi - PR China.
2 Department of Otorhinolaryngology, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi - PR China.
Tumori. 2018 Jan-Feb;104(1):37-42. doi: 10.5301/tj.5000706.
Although high expression of ZBTB7A is positively relative to metastasis in nasopharyngeal carcinoma (NPC) patients, the association between its low expression and metastasis of NPC remains unclear. The present study aimed to definitely identify the association.
The level of ZBTB7A was effectively knocked down by stable transfection of short hair RNA plasmid in NPC cell lines CNE2 and 5-8F (shRNA-CNE2 and shRNA-5-8F), compared with the cells that stably transfected empty plasmid (NC-CNE2 and NC-5-8F). The levels of ZBTB7A were assessed by real-time polymerase chain reaction and Western blot in the cell lines. MTT assay, colorimetric focus-formation assay, flow cytometry, wound healing assay, transwell assays, and xenograft model were performed to analyze cell vitality, proliferation, cell cycle, migration, invasion, and tumorigenicity.
The levels of ZBTB7A were effectively reduced in shRNA-CNE2 and shRNA-5-8F. Their carcinogenicity was stronger separately than the abilities of NC-CNE2 and NC-5-8F. NC-CNE2 and shRNA-CNE2 were selected to establish the xenograft model because of their stronger tumorigenicity than NC-6-10B and shRNA-5-8F. The assay showed that shRNA-CNE2 had stronger tumorigenicity than NC-CNE2.
The results demonstrated the reverse association between the expression of ZBTB7A and the tumorigenicity of NPC. We postulate that some oncogenic pathways, which are suppressed by ZBTB7A, will vicariously promote the proliferation and progression of NPC when ZBTB7A is decreased.
尽管ZBTB7A的高表达与鼻咽癌(NPC)患者的转移呈正相关,但其低表达与NPC转移之间的关联仍不清楚。本研究旨在明确确定这种关联。
通过在NPC细胞系CNE2和5-8F中稳定转染短发夹RNA质粒(shRNA-CNE2和shRNA-5-8F)有效敲低ZBTB7A水平,与稳定转染空质粒的细胞(NC-CNE2和NC-5-8F)进行比较。通过实时聚合酶链反应和蛋白质免疫印迹法评估细胞系中ZBTB7A的水平。进行MTT法、比色集落形成法、流式细胞术、伤口愈合试验、Transwell试验和异种移植模型分析细胞活力、增殖、细胞周期、迁移、侵袭和致瘤性。
shRNA-CNE2和shRNA-5-8F中ZBTB7A水平有效降低。它们的致癌性分别比NC-CNE2和NC-5-8F更强。由于NC-CNE2和shRNA-CNE2比NC-6-10B和shRNA-5-8F具有更强的致瘤性,因此选择它们建立异种移植模型。试验表明,shRNA-CNE2比NC-CNE2具有更强的致瘤性。
结果表明ZBTB7A表达与NPC致瘤性之间呈负相关。我们推测,一些被ZBTB7A抑制的致癌途径,当ZBTB7A降低时,将间接促进NPC的增殖和进展。