利用cDNA-AFLP方法在体内鉴定结直肠癌中新型TGIF2LX靶基因
In vivo identification of novel TGIF2LX target genes in colorectal adenocarcinoma using the cDNA-AFLP method.
作者信息
Mobini Gholam Reza, Ghafari Arefeh, Amanpour Saeid, Fateh Roohollah, Ghahremani Mohammad Hossein, Muhammadnejad Samad, Dehpour Ahmad Reza, Akbari Abolfazl, Hoseiniharouni Seyed Mojtaba, Kazemirad Elham, Bolhassani Manzar, Heidari Mansour
机构信息
Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran; Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Department of Medical Genetics, Tehran University of Medical Sciences, Tehran, Iran.
出版信息
Arab J Gastroenterol. 2018 Jun;19(2):65-70. doi: 10.1016/j.ajg.2018.05.001. Epub 2018 Jun 28.
BACKGROUND AND STUDY AIMS
Homeobox-containing genes are composed of a group of regulatory genes encoding transcription factors involved in the control of developmental processes. The homeodomain proteins could activate or repress the expression of downstream target genes. This study was conducted to in vivo identify the potential target gene(s) of TGIF2LX in colorectal adenocarcinoma.
METHODS
A human colorectal adenocarcinoma cell line, SW48, was transfected with the recombinant pEGFPN1-TGIF2LX. The cells were injected subcutaneously into the flank of the three groups of 6-week-old female athymic C56BL/6 nude mice (n = 6 per group). The transcript profiles in the developed tumours were investigated using the cDNA amplified fragment length polymorphism (cDNA-AFLP) technique.
RESULTS
The real-time RT-PCR and DNA sequencing data for the identified genes indicated that the N-terminal domain-interacting receptor 1 (Nir1) gene was suppressed whereas Nir2 and fragile histidine triad (FHIT) genes were upregulated followed by the overexpression of TGIF2LX gene.
CONCLUSION
Downregulation of Nir1 and upregulation of Nir2 and FHIT genes due to the overexpression of TGIF2LX suggests that the gene plays an important role as a suppressor in colorectal adenocarcinoma.
背景与研究目的
含同源框基因由一组调控基因组成，这些基因编码参与发育过程控制的转录因子。同源结构域蛋白可激活或抑制下游靶基因的表达。本研究旨在体内鉴定TGIF2LX在结直肠癌中的潜在靶基因。
方法
用人重组pEGFPN1-TGIF2LX转染人结直肠腺癌细胞系SW48。将细胞皮下注射到三组6周龄雌性C56BL/6无胸腺裸鼠（每组n = 6）的胁腹。使用cDNA扩增片段长度多态性（cDNA-AFLP）技术研究所形成肿瘤中的转录谱。
结果
对鉴定出的基因进行实时RT-PCR和DNA测序数据表明，N端结构域相互作用受体1（Nir1）基因被抑制，而Nir2和脆性组氨酸三联体（FHIT）基因在TGIF2LX基因过表达后上调。
结论
由于TGIF2LX过表达导致Nir1下调以及Nir2和FHIT基因上调，提示该基因在结直肠癌中作为抑癌基因发挥重要作用。
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