國立臺灣大學醫學院外科陳炯年2006-07-262018-07-112006-07-262018-07-112005-07-31http://ntur.lib.ntu.edu.tw//handle/246246/24562p40/ EBP2/ NoBP 基因最早是發現自癌細胞中的細胞核蛋白，將它轉送到老鼠細胞中發現 可以促進細胞的增生。在酵母菌中它參與了rRNA 和核醣體的合成，在動物細胞中它可以 與鼻咽癌病毒蛋白EBNA1 結合，幫助病毒DNA 在細胞內的維持。 先前我們在研究在不同血管密度的胃癌組織裡基因表現的差異時，發現的p40/ EBP2/ NoBP 基因表現在胃癌組織中。為了探討p40/ EBP2/ NoBP 是否與胃癌的發生和血管新生之間的 關係，我們在北方墨點實驗結果發現這基因在有些胃癌組織比正常組織中表現要高；且此 基因的表現在已分化成微血管的內皮細胞是受到抑制，在免疫細胞分化過程中，在不具複 製能力的已分化細胞也有相同的現象。我們以具有抑制血管新生的藥物15-d-PGJ2 刺激的內 皮細胞，也發現p40/ EBP2/ NoBP 基因表現受到抑制。 根據先前文獻的發現以及我們實驗室的研究結果，在本計劃中有四個主要研究目的：第一 個目的是要探討15-d-PGJ2 抑制p40/ EBP2/ NoBP 基因表現之機制。p40/ EBP2/ NoBP 基因 啟動子上有許多Sp1 和c-Ets-1 轉錄因子結合位，15-d-PGJ2 對血管新生的抑制作用是否是 藉由抑制內皮細胞Sp1 和c-Ets-1 的結合能力，而間接抑制p40/ EBP2/ NoBP 的表現，使得 內皮細胞增生受到抑制。 第二個目的是要研究p40/ EBP2/ NoBP 調控細胞生長之機制。在動物細胞中，p40/ EBP2/ NoBP 基因的高度表現可以促進細胞的生長，但機制仍不清楚。且在不同物種之間此基因 有很高的相似性。所以我們推論在不同物種細胞中，此基因可能以相同的機制調控細胞的 生長。我們計劃以免疫沉澱技術來找到與它結合在一起的蛋白。 第三個目的目是探討p40/ EBP2/ NoBP 基因表現與胃癌發生之關係。EBV 病毒與鼻咽癌和 淋巴癌有一定的相關性。在胃癌中的Gastric lymphoepithelioma- like carcinoma 幾乎也都有 受到EBV 病毒的感染。鼻咽癌病毒又可藉著EBNA1 和p40/ EBP2/ NoBP 的結合，將病毒 DNA 穩定的保存在上皮細胞裡。所以我們想要了解此基因是否在Gastric lymphoepithelioma- like carcinoma 形成過程中扮演重要的角色。 第四個目的是探討p40/ EBP2/ NoBP 基因是否為一前趨致癌基因(proto-oncogen)。 在我們的軟洋菜膠實驗中(Soft agar assay)發現它雖不能夠在形成細胞群落，但仍有能力在 軟洋菜膠培養基上進行二到四次的細胞分裂。所以我們計劃將它與ras 一起送到細胞內，以 觀察加上ras 基因的表現是否就可以幫助3T3 跨越細胞癌化的門檻。 以上的研究計劃成果，可以幫助我們進一步了解p40/ EBP2/ NoBP 基因在細胞生長調控、 胃癌發生和血管新生過程中所扮演的角色。The p40/ EBP2/ NoBP was firstly discovered from the nucleolus of cancer cell. Transfect this gene into animal cell could promote cell proliferation. In budding yeast, the gene function is essential for rRNA and ribosome synthesis. In EBV infected cell, it could interact with viral nuclear protein EBNA1 to maintain viral genome during cell division. In our previous study of gene expression profile of gastric cancers with different vessel indexes, we discovered that p40/ EBP2/ NoBP expressed in gastric cancer tissue. In order to identify the relationship of p40/ EBP2/ NoBP with gastric cancer process and angiogenesis, In Northern blotting assay, we had found that p40/ EBP2/ NoBP overexpressed in some gastric cancer tissues. And its expression is downregulated in capillary-like HUVEC cultured on Matrixgel. In monocyte –macrophage differentiation process, p40/ EBP2/ NoBP is also downregulted in differentiated macrophage. In gene expression study of HUVEC treated with 15-d-PGJ2, a antiangiogenesis drug, we had found that p40/ EBP2/ NoBP was also inhibited。 According to the previous publication and our findings, there are four major objects in this project. The first object is to identify the downregulation mechanism of p40/ EBP2/ NoBP in HUVEC treated with 15-d-PGJ2. There are 8 Sp1 binding sites and 4 c-Ets-1 binding sites in promoter region of p40/ EBP2/ NoBP (-500 to +1). We hypothesize that 15-d-PGJ2maybe inhibit the binding activity of Sp1and c-Ets-1, moreover, doweregulate p40/ EBP2/ NoBP expression. Finally, the endothelial cell proliferation is stopped. The secondary object is to study the mechanism of p40/ EBP2/ NoBP in cell growth control. There are many p40/ EBP2/ NoBP. homologues found in different species. And, this gene overexpression can promote animal cell proliferation, but, the mechanism still unknown. We suggest that the cell growth mechanism controlled by p40/ EBP2/ NoBP. maybe be similar in different species. We will utilize immunoprecipitation assay of p40/ EBP2/ NoBP to identify its interaction protein for its function study. The third object is to evaluate the relationship between p40/ EBP2/ NoBP expression and EBV infected gastric cancer. EBV infection is related to nasopharyngeal cacinoma, Burkitt’s lymphoma. There are publication reported that gastric lymphoepithelioma- like carcinoma almost had been infected by EBV. In EBV infected cells, viral protein EBNA1 can interaction with p40/ EBP2/ NoBP to help viral genome separation during cell division. So, we hypothesize that p40/ EBP2/ NoBP should play an important role in EBV infected gastric cancer. The forth object is to identify whether p40/ EBP2/ NoBP is a novel proto-oncogene. Our previous data shown that p40/ EBP2/ NoBP transfected 3T3 cell although can’t form obvious colony in soft agar assay, but they can divided 2 to 4 times, then stop growth. We plan to cotransfect 3T3 with p40/ EBP2/ NoBP and ras, to monitor whether they can form colony. We predict these results can identify the roles of p40/ EBP2/ NoBP in gastric cancer process and angiogenesis.application/pdf1455210 bytesapplication/pdfzh-TW國立臺灣大學醫學院外科p40/ EBP2/ NoBPPPAR-γ胃癌血管新生細胞增生腫瘤發生基因調控gastric cacerangiogenesiscell proliferationtumorigenesisgene control[SDGs]SDG3reporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/24562/1/932314B002270.pdf