NAIF1对胃癌细胞中IFIT家族蛋白的影响

张巧; 黄常志; 祝建疆; 李燕; 赵玫; 李可心; 李东东; 戚红; 黄声凯

doi:10.3971/j.issn.1000-8578.2022.21.0551

NAIF1对胃癌细胞中IFIT家族蛋白的影响

Effect of NAIF1 on IFIT Family Proteins in Gastric Cancer

摘要

摘要:
目的探讨核诱导凋亡因子NAIF1在胃癌细胞系MKN45中的功能。
方法构建带有Tet-on系统的质粒pLVX-Tight-Flag-NAIF1-puro，在胃癌细胞系MKN45中，加入四环素衍生物强力霉素（Dox）诱导NAIF1表达，分别在加入Dox之后0、6、12、24 h收取细胞进行基因表达谱芯片检测，对差异表达的基因进行生物学分析，并利用实时荧光定量PCR（qRT-PCR）和Western blot技术验证筛选的目的差异基因在mRNA和蛋白水平上的变化。
结果基因芯片测定结果显示干扰素诱导蛋白（IFIT）家族基因分子IFIT1、IFIT2和IFIT3在第24 h表达量明显增加，qRT-PCR同样表现出该变化，Western blot在蛋白水平进一步验证了该变化。IFIT家族另一成员IFIT5在mRNA和蛋白水平表达量的变化趋势并不明显。
结论胃癌细胞中过表达NAIF1能够促进免疫系统相关的部分干扰素诱导蛋白（IFIT）家族蛋白的表达。

Abstract:
Objective To investigate the effect of NAIF1 in gastric cancer cell lines MKN45.
Methods We constructed pLVX-Tight-Flag-NAIF1-puro plasmid with Tet-on system. DOX was added to induce NAIF1 expression in MKN45 cells. The cells were collected at 0, 6, 12 and 24 hours after DOX addition for gene expression microarray detection and biological analysis of differentially expressed genes. qRT-PCR and Western blot were used to verify the changes in mRNA and protein levels of the selected target differential genes.
Results The biological analysis of gene microarray hybridization results showed that IFIT1, IFIT2 and IFIT3 expression significantly increased at 24h, qRT-PCR also showed this change, and Western blot further verified the change in protein level. However, IFIT5 showed no significant change in mRNA and gene expression.
Conclusion Over-expression of NAIF1 in gastric cancer cells can promote the expression of some immune system-related IFIT family proteins.

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