Ziaei Saeid, Rezaei-Tavirani Mostafa, Ardeshirylajimi Abdolreza, Arefian Ehsan, Soleimani Masoud
Department of Basic Sciences, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Student Research Committee, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Galen Med J. 2019 Aug 10;8:e1308. doi: 10.31661/gmj.v8i0.1308. eCollection 2019.
THAP11 is a recently discovered pluripotency factor and described as an important gene that involved in embryonic stem cells self-renewal and embryo development, which works independently with other known pluripotency factors. We aimed to overexpressed the THAP11 gene in primary fibroblast cells to determine the effects of the THAP11 on these cells.
The THAP11 gene was amplified using PCR followed by ligation into pCDH vector and lentiviral particle production in HEK293T cells by using psPAX2 and pMD2.G helper vectors. The human fibroblast cells were transduced using viral particles and after confirmation overexpression, the key pluripotency factors were estimated using real-time PCR and changes in proliferation rate was measured by the 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium bromide (MTT) test.
The overexpression of THAP11 in fibroblast cells leads to increase the expression level of , , and as key pluripotency genes and a decrease in proliferation rate according to MTT results.
Our results confirm that we are faced with a molecule with double features, which could be involved in pluripotency and proliferation suppressor simultaneously. It seems that the roles of THAP11 in pluripotency are so complex and attributed to other regulatory molecules.
THAP11是最近发现的一种多能性因子,被描述为参与胚胎干细胞自我更新和胚胎发育的重要基因,它独立于其他已知的多能性因子发挥作用。我们旨在使原代成纤维细胞中的THAP11基因过表达,以确定THAP11对这些细胞的影响。
使用PCR扩增THAP11基因,然后将其连接到pCDH载体中,并通过使用psPAX2和pMD2.G辅助载体在HEK293T细胞中产生慢病毒颗粒。使用病毒颗粒转导人成纤维细胞,在确认过表达后,使用实时PCR估计关键多能性因子,并通过3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐(MTT)试验测量增殖率的变化。
根据MTT结果,成纤维细胞中THAP11的过表达导致关键多能性基因、、和的表达水平增加,增殖率降低。
我们的结果证实,我们面对的是一种具有双重特性的分子,它可能同时参与多能性和增殖抑制。THAP11在多能性中的作用似乎非常复杂,并且归因于其他调节分子。