Qu Yang-Hua, Fu Jun-Cai, Liu Kun, Zuo Zhao-Yun, Jia Hui-Na, Ma Yong, Luo Hai-Ling
State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.
Int J Mol Sci. 2016 Jun 27;17(7):1016. doi: 10.3390/ijms17071016.
α-Tocopherol transfer protein (α-TTP) is a ~32 kDa protein expressed mainly in hepatocytes. The major function of the protein is to bind specifically to α-tocopherol and, together, the complex transfers from late lysosomes to the cell membrane. A previous study indicated that some factors might be required in the transferring process. However, there is little information available about the potential transferring factors. In addition, there remains much to learn about other physiological processes which α-TTP might participate in. Thus, in this study a human α-TTP eukaryotic expression vector was successfully constructed and expressed in human hepatoma cells (HepG2). The sensitive genes related to α-TTP were then screened by microarray technology. Results showed that expression of the vector in HepG2 cells led to the identification of 323 genes showing differential expression. The differentially expressed transcripts were divided into four main categories, including (1) cell inflammation; (2) cell cycle and cell apoptosis; (3) cell signaling and gene regulation; and (4) cellular movement. A few cellular movement related transcripts were selected and verified by quantitative real-time PCR. Expressions of some were significantly increased in α-TTP-expressed group, which indicated that these factors were likely to play a role in the transferring process.
α-生育酚转运蛋白(α-TTP)是一种主要在肝细胞中表达的约32 kDa的蛋白质。该蛋白质的主要功能是特异性结合α-生育酚,并且该复合物一起从晚期溶酶体转移至细胞膜。先前的一项研究表明,在转移过程中可能需要一些因子。然而,关于潜在的转移因子的信息很少。此外,关于α-TTP可能参与的其他生理过程仍有很多需要了解的。因此,在本研究中成功构建了人α-TTP真核表达载体并在人肝癌细胞(HepG2)中表达。然后通过微阵列技术筛选与α-TTP相关的敏感基因。结果表明,该载体在HepG2细胞中的表达导致鉴定出323个显示差异表达的基因。差异表达的转录本分为四大类,包括(1)细胞炎症;(2)细胞周期和细胞凋亡;(3)细胞信号传导和基因调控;以及(4)细胞运动。选择了一些与细胞运动相关的转录本并通过定量实时PCR进行验证。在α-TTP表达组中,一些转录本的表达显著增加,这表明这些因子可能在转移过程中起作用。