DPH1基因新可变剪接体DPH1-IR在鼻咽癌中致癌功能及机制

项目名称： DPH1基因新可变剪接体DPH1-IR在鼻咽癌中致癌功能及机制

项目编号： No.81460430

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 李思维

作者单位： 桂林医学院

项目金额： 47万元

中文摘要： DPH1是卵巢癌抑癌基因。我们前期在人鼻咽癌细胞株C666-1中发现并克隆了DPH1基因一个新的可变剪接体（DPH1-IR），该可变剪接体在10号及11号外显子之间保留了一个内含子，但其功能及机制不清。我们发现：DPH1-IR mRNA在人鼻咽癌细胞、组织中表达上调。据此,我们提出DPH1-IR可能具有潜在致癌功能的假设。为证实该假设，我们在DPH1-IR低表达的鼻咽癌细胞HONE1中转染DPH1野生型及DPH1-IR基因，发现：DPH1-IR能表达蛋白，与野生型相反，DPH1-IR能增强细胞克隆及侵袭能力。为此，我们拟研制DPH1-IR特异性抗体，并通过PCR、免疫组化等方法进一步确认该剪接体表达的临床意义，继而探讨它对鼻咽癌细胞侵袭能力、裸鼠成瘤及转移能力等影响。在此基础上，进一步研究该剪接体的致癌机制。旨在明确DPH1新剪接体在鼻咽癌发生、发展中的作用及意义，并初步阐明其分子机制。

中文关键词： C03_鼻咽肿瘤；可变剪接；DPH1；基因表达

英文摘要： Ovarian cancer gene 1(DPH1) functions as a tumor suppressor gene in ovarian cancer. Our previous studies successfully found and cloned a new alternative splicing transcript(named DPH1-IR) of DPH1 gene in C666-1, a nasopharyngeal carcinoma (NPC) cell strain. One single intron between Exon 10 and Exon 11 of DPH1 pre-mRNA was retained in transcription and produced a novel transcript. But its functions and mechanisms in NPC are unknown.We detected that expression of DPH1-IR in mRNA level was higher in NPC cell lines and tissue speciems than in normal cantrols. On basis of these results，we hypothesized that DPH1-IR probably demonstrated functions of potential oncogene. To validate the hypothesis, we constructed plasmids of DPH1-IR and DPH1-WT and transfected them into HONE1, a nasopharyngeal cancer cell with low DPH1-IR-expressing level,which was proved by our preliminary foundings, respectively. Surprisingly, we found that DPH1-IR could express protein and increase the clone numbers in clone formation assay whereas DPH1-WT could dramatically decrease the clone numbers.Fist,we will develop a specific DPH1-IR antibody,and confirm the clinical significance of the new alternative splicing transcript by using the methods of polymerase chain reaction (PCR)and immunohistochemical staining.Next, we will overexpress and knockdown DPH1-IR in NPC cell lines, and study the abilities of invasive and the clone formation in vitro, and also study abilities of both tumorgenesis and tumor metastasis in nude mice mode. Finally,The oncogenic mechanism of DPH1-IR in NPC will also be explored. The target is to demenstrate the function and significance of the new alternative splicing transcript of DPH1 gene in the process of formation and development of NPC,and preliminarily explore its oncogenic mechanisms.

英文关键词： nasopharyngeal carcinoma;alternative splicing;DPH1;gene expression