Identification and analysis of the promoter region of the STGC3 gene
Identification and analysis of the promoter region of the STGC3 gene
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DOI：
10.5114/aoms.2015.49213
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发表时间：
2015-01-01
影响因子：
3.8
通讯作者：
Zhang, Zhiwei
中科院分区：
文献类型：
--
作者：
Li, Suyun;Wang, Lili;Zhang, Zhiwei
简介：鼻咽癌（NPC）是头颈部常见的恶性肿瘤。 STGC3基因与鼻咽癌的发生有关。本研究的目的是探索STGC3基因的启动子区域。材料与方法：应用生物信息学技术预测其启动子区域，构建基因启动子区域荧光素酶基因载体并转染人胚肾上皮293T细胞系、人鼻咽癌CNE2细胞系和永生化鼻咽癌。 上皮NP69细胞系。将重组质粒pGL3-en283、pGL3-en281、pGL3-en571、空质粒pGL3-control、阴性对照pGL3-enhance和海洋肠荧光素酶表达载体pRL-SV40内参转染NP69细胞、293T细胞和CNE2细胞。双荧光素酶活性检测显示荧光素酶发光值和海洋肠荧光素酶发光值。计算每个细胞中的相对荧光素酶活性（RLA）。结果：与阴性对照 pGL3-enhance 质粒相比，我们在 NP69、293T 和 CNE2 细胞中观察到质粒 pGL3-en283、pGL3-en281 和 pGL3-en571 具有较强的启动子活性。其中，pGL3-en281表现出最强的启动子活性，这三种重组质粒在293T细胞中表现出比CNE2细胞更强的启动子活性。结论：pGL3-en281质粒在三种细胞中表现出比pGL3-en571更强的启动子活性，表明-11048 bp至-653 bp可能是核心启动子区域。
Introduction: Nasopharyngeal carcinoma (NPC) is a common malignant tumor of the head and neck. The STGC3 gene is related to development of nasopharyngeal cancer. The aim of this study is to explore the promoter region of the STGC3 gene.Material and methods: The bioinformatic technique was applied to predict its promoter region and construct the gene promoter region luciferase for the gene vector and transfection of the human embryonic kidney epithelial 293T cell line, human nasopharyngeal carcinoma CNE2 cell line and immortalized nasopharyngeal epithelial NP69 cell line. The recombinant plasmid pGL3-en283, pGL3-en281, pGL3-en571, empty plasmid pGL3-control, negative control pGL3-enhance and internal control of marine intestine luciferase expression vector pRL-SV40 were transfected into NP69 cells, 293T cells and CNE2 cells. Dual luciferase activity detection showed luciferase luminescence values and marine intestine luciferase luminescence values. Relative luciferase activity (RLA) in each cell was calculated.Results: We observed strong promoter activity of plasmid pGL3-en283, pGL3-en281 and pGL3-en571 in NP69, 293T and CNE2 cells compared with the negative control pGL3-enhance plasmid. Among them, pGL3-en281 showed the strongest promoter activity, and these three kinds of recombinant plasmids showed stronger promoter activity in 293T cells than in CNE2 cells.Conclusions: The pGL3-en281 plasmid showed stronger promoter activity than pGL3-en571 in the three cells, indicating that -11048 bp to -653 bp might be the core promoter region.