EBV高表达miR-BART8激活AKT/FOXO3a及Ras/ERK信号通路介导鼻咽癌细胞EMT转化及放化疗抵抗的机制研究

Using microarray analysis we previously found that EBV-miR-BART8 was highly expressed in nasopharyngeal carcinoma (NPC)and 20.12 times of normal nasopharyngeal tissues,indicating that EBV-miR-BART8 may be a potential regulatory factor for nasopharyngeal carcinoma.In order to verify the hypothesis, firstly we will collect clinical samples and confirm that EBV-miR-BART8 is related to the stage and prognosis of nasopharyngeal carcinoma.Next, in vivo and in vitro experiments and morphological observation will be used to identify EBV-miR-BART8 can inhibit the apoptosis of nasopharyngeal carcinoma cells, promote metastasis and epithelial mesenchymal transition.Further, according to the clinical specimen analysis, in situ hybridization, microarray analysis, western blot, confocal and other experiments will be used to explore the host genes, signaling pathways and downstream molecules of EBV-miR-BART8.Finally, we will use animal models to confirm EBV-miR-BART8 could induce resistance to radiotherapy and chemotherapy of nasopharyngeal carcinoma cells, and silencing EBV-miR-BART8 expression could inhibit the malignant progression of nasopharyngeal carcinoma cells.This study will reveal the mechanism of EBV miRNA in promoting the metastasis and recurrence of nasopharyngeal carcinoma, and provide scientific evidence of EBV-miR-BART8 as a new target for the treatment of nasopharyngeal carcinoma.

前期芯片分析我们发现EBV编码的miR-BART8在鼻咽癌组织中明显高表达，为正常鼻咽组织的20.12倍，提示EBV-miR-BART8可能是鼻咽癌的潜在调控因子。为验证假说，本项目首先扩大临床样本量，证实EBV-miR-BART8与鼻咽癌分期及预后相关。接下来，利用体内外功能实验及形态观察明确EBV-miR-BART8促进转移及EMT转化。进一步，我们利用芯片分析、western blot、confocal等实验结合临床标本分析，从机制角度探究EBV-miR-BART8调控的宿主靶基因及相关下游信号分子。最后，构建动物治疗模型，从治疗角度揭示EBV-miR-BART8能够诱导鼻咽癌细胞放化疗抵抗，沉默EBV-miR-BART8表达能抑制鼻咽癌细胞恶性进展。本研究将有可能揭示EBV miRNA促进鼻咽癌转移及复发的新机制，为确立EBV-miR-BART8作为鼻咽癌治疗的新靶点提供科学依据。

前期芯片分析我们发现EBV编码的miR-BART8-3p在鼻咽癌组织中明显高表达，为正常鼻咽组织 的20.12倍，提示miR-BART8-3p可能是鼻咽癌的潜在调控因子。为验证假说，本项目首先扩大 临床样本量，证实EBV-miR-BART8与鼻咽癌分期及预后相关。接下来，我们首先构建稳定过表达miR-BART8-3p的鼻咽癌细胞株，利用体内外实验明确EBV-miR-BART8-3p通过调节ATM/ATR信号通路促进鼻咽癌的放疗抵抗；利用RNA测序分析筛选出miR-BART8-3p潜在的宿主靶基因PAG1，并表明miR-BART8-3p对PAG1的结构和功能均有调控作用，同时进一步体内外实验明确miR-BART8-3p靶向抑制PAG1的表达促进鼻咽癌细胞转移和EMT转化；最后结合临床样本分析，明确miR-BART8-3p可能是重要的预后因子。在本课题的资助下，课题申请者分别以第一作者发表SCI论文1篇，以末位通讯作者发表 SCI 论文4 篇，其中II区论文2篇，包括：Oncogene 2021.( IF:9.867)、J Cell Physiol 2021(IF:6.383)；以第一发明人发表国家发明专利1项。本研究将有可能揭示EBV miRNA促进鼻咽癌转移及复发的新机制，为确立miR-BART8-3p作为鼻咽癌治疗的新靶点提供科学依据。