Akbari Abolfazl, Agah Shahram, Heidari Mansour, Mobini Gholam Reza, Faghihloo Ebrahim, Sarveazad Arash, Mirzaei Alireza
Colorectal Research Center, Iran University of Medical Sciences, Tehran, Iran. Email:
Asian Pac J Cancer Prev. 2017 Aug 27;18(18):2101-2108. doi: 10.22034/APJCP.2017.18.8.2101.
Background: TGIF2LX (transforming growth factor beta-induced factor 2 like, X-linked) is a homeodomain (HD) protein that has been implicated in the negative regulation of cell signaling pathways. The aim of this study was to investigate the possible functions of TGIF2LX in colon adenocarcinoma cells. Methods: The human SW48 cell line was transfected with cDNA for the wild-type TGIF2LX gene and gene/protein over-expression was confirmed by microscopic analysis, real time RT-PCR and Western blotting techniques. In vitro cell proliferation was evaluated by MTT and BrdU assays. After developing a colon tumor model in nude mice, immunohistochemical (IHC) staining of tumor tissue was carried out for Ki-67 (proliferation) and CD34 (angiogenesis) markers. To predict potential protein partners of TGIF2LX, in-silico analysis was also conducted. Results: Obtained results showed over-expression of TGIF2LX as a potential transcription factor could inhibit either proliferation or angiogenesis (P<0.05) in colon tumors. In-silico results predicted interaction of TGIF2LX with other proteins considered important for cellular development. Conclusions: Our findings provided evidence of molecular mechanisms by which TGIF2LX could act as a tumor suppressor in colon adenocarcinoma cells. Thus, this gene may potentially be a promising option for colon cancer gene-based therapeutic strategies.
TGIF2LX(X连锁的转化生长因子β诱导因子2样蛋白)是一种同源结构域(HD)蛋白,与细胞信号通路的负调控有关。本研究的目的是探讨TGIF2LX在结肠腺癌细胞中的可能功能。方法:用野生型TGIF2LX基因的cDNA转染人SW48细胞系,并通过显微镜分析、实时RT-PCR和蛋白质印迹技术确认基因/蛋白的过表达。通过MTT和BrdU试验评估体外细胞增殖。在裸鼠中建立结肠肿瘤模型后,对肿瘤组织进行免疫组织化学(IHC)染色,检测Ki-67(增殖)和CD34(血管生成)标记物。为了预测TGIF2LX的潜在蛋白伴侣,还进行了计算机模拟分析。结果:获得的结果表明,TGIF2LX作为一种潜在的转录因子过表达可抑制结肠肿瘤的增殖或血管生成(P<0.05)。计算机模拟结果预测TGIF2LX与其他对细胞发育重要的蛋白质相互作用。结论:我们的研究结果提供了TGIF2LX可作为结肠腺癌细胞中肿瘤抑制因子的分子机制的证据。因此,该基因可能是基于基因的结肠癌治疗策略的一个有前景的选择。