摘要
目的通过构建N-乙酰转移酶基因(NAA10)腺病毒表达载体,探讨其对食管癌细胞TE-1增殖、细胞周期和凋亡的影响。方法 PCR技术扩增NAA10全长编码基因,通过重组腺病毒表达系统将其克隆入pShuttle-IRES-hrGFP-1穿梭载体,重组腺病毒表达载体并包装腺病毒颗粒Ad-NAA10。将携带人NAA10基因的腺病毒感染食管癌细胞TE-1,Western blot检测NAA10蛋白的表达,CCK-8检测其对TE-1细胞增殖的影响,碘化丙啶(PI)染色检测其对TE-1细胞周期的影响,Hoechst/PI双染检测其对TE-1细胞凋亡的影响。结果酶切鉴定和测序证实NAA10腺病毒表达载体构建成功,腺病毒颗粒Ad-NAA10能在TE-1细胞中正常表达;NAA10可以抑制TE-1细胞的增值,影响细胞周期分布,促进细胞凋亡。结论 NAA10通过G1期阻滞和诱导凋亡从而抑制食管癌细胞的增殖,为进一步确定食管癌特异性生物标志物及探讨食管癌发病机制奠定基础。
Objective To investigate the effect of NAA10 on cell proliferation,cell cycle and apoptosis in TE-1 cells by constructing a NAA10 adenovirus expression vector.Methods NAA10 cDNA was amplified by RT-PCR,and was then cloned into pShuttle-IRES-hrGFP-1 shuttle carrier by recombinant adenovirus expressing system.Recombinant adenovirus expression vector and packing gland virus particles Ad-NAA10.NAA10 gene adenovirus was then transfected into TE-1 cells,and Western blot was used to detect NAA10 expression level.the TE-1 cells proliferation was analyzed by CCK8,and cell cycle was analyzed by propyl iodide（PI）organism.Hoechst/PI double dye was used to test its effect on cell apoptosis.Results Enzyme identification and sequencing confirmed that the NAA10 adenovirus expression vector was successfully constructed,and Ad-NAA10 adenovirus particles can normally present in TE-1 cells.NAA10 can inhibit cell proliferation,affect the distribution of cell cycle,and promote cell apoptosis.Conclusion NAA10 can inhibit cell proliferation by G1 arrest and inducing apoptosis.Furthermore,this study lays down the foundation for the specific biological markers and for exploring the pathogenesis of esophageal cancer.
出处
《福建医药杂志》
CAS
2017年第4期77-80,共4页
Fujian Medical Journal
基金
福建省省属公益类科研院所基本科研专项(2014R1031-6
2015R1031-7)
福建省卫生计生青年骨干人才培养项目(2016-ZQN-22)
福建省卫生计生青年科研课题(2017-1-28)
