林中天2006-07-262018-07-092006-07-262018-07-092003http://ntur.lib.ntu.edu.tw//handle/246246/28723乳癌對人類是重要且常見的腫瘤，其產生 的病因是複雜為多因子牽涉的結果。Secreted frizzled related proteins (sFRPs)是最近被 發現與Wnt-Frizzled 訊息傳遞傳導路徑的調節 和細胞凋亡(apoptosis)的調節中扮演著雙重 角色的蛋白質。我們實驗室最近發現sFRP2 基 因在人類與犬乳癌中有大量的表現及活化，但 是在正常乳腺組織中則沒有表現。我們為了有 系統地進一步研究隻乳癌中的sFRP2 在功能上 的角色與腫瘤分子生物學上的機制，擬定了下 列的幾項研究策略。 這個計畫原預定包括了六個主要的部分， 需要3 年的研究時間，但最終僅獲核給一年計 劃經費，在這一年主要的工作在於建立並分析 臺灣地區早發性乳癌組織之sFRP2 基因表現， 並建立早發性乳癌的初代培養(primary culture)細胞株並且純化、分析乳腺上皮細 胞。這一年我們已成功地分析早發性乳癌組織 的sFRP2 基因之表現。並建立及分析數個本地 病例之早發性乳癌的初代培養細胞株。這些細 胞利用下列技術分析其特性，包括反轉錄鏈聚 合脢反應(RT-PCR) 、原位雜交法( in situ hybridization) 與免疫化學染色 (immunohistochemistry) 法偵測sFRP2 的表 現。結果發現sFRP2 基因之mRNA 及蛋白質在乳 癌組織大量且顯著高比例之表現(86% mRNA 表現 陽性)，然而在正常乳腺組織則顯著較低比例之 表現(43% mRNA 表現陽性)。未來在下一個階段， sFRP2 將被轉殖入含有GFP 基因與CMV 啟動子的 哺乳類細胞表現載體，藉由lipofection 方式 將GFP-sFRP2 穩定地轉染入(transfect) 早發 性乳癌的初代培養細胞株，以進行更進一步的 sFRP2 功能分析。 本階段之研究結果，預期將提供重要之學 術資訊，以了解sFRP 基因族在人早發性乳癌 之表現情形。此外，此計劃也為未來下一階段 進一步研究sFRP 基因族不同成員之各種功 能，及了解早發性乳癌複雜之病因，提供進一 步研究分析之基礎。Breast cancers are one of the most important and common tumors in humans. The etiology of breast cancers is complex. The secreted frizzled related proteins (sFRPs) are newly identified proteins and implicated to have dual roles of modulation of Wnt-Frizzled signal transduction pathway and regulation of apoptosis. We have recently found that sFRP2 was expressed abundantly in human breast cancer (mostly late onset) and canine mammary gland tumors (MGT) tissues but was undetectable in normal mammary glands. To systematically investigate the functional roles and molecular mechanisms of sFRP2 in early onset breast cancers, several strategies are to be carried out as described below. The project was intended to be comprised of six major parts for a period of 3 years. However, only one year of funding was granted. During this year, breast cancer tissues and primary cell cultures from native early onset breast cancer tissues have been established and purified for epithelial cells. We have successfully analyzed the sFRP2 expression in early onset breast cancer tissues. In addition, we have established and analyzed native primary early onset breast cancer cell lines from surgically excised breast cancer specimens. The cells are characterized for their cell origins, expression of sFRP2 by RT-PCR, in situ hybridization, and immunohistochemistry. Expression experiments revealed the sFRP2 was abundantly expressed with significantly a higher expression rate in early onset breast cancer tissues (86% mRNA positive), while much lower expression rate in normal breast tissues (43% mRNA positive). In the following step, human sFRP2 is cloned into a mammalian expression vector with GFP reporter gene and CMV promoter. The GFP-sFRP2 is stably transfected into primary early onset breast cancer cell lines by lipofection for further analysis from the next stage of the project although no funding is granted for the next year’s study. The results of this stage of the project should offer important scientific basis and information to understand the roles of sFRP gene family in early onset breast cancers. It also provides a basis for further analysis of functions of different members of the sFRP gene family and elucidation of the complex etiology of early onset breast cancers.application/pdf8829720 bytesapplication/pdfzh-TW國立臺灣大學獸醫學系暨研究所分泌性細胞凋亡基因分泌性frizzled蛋白基因基因表現基因轉染早發性乳癌secreted apoptosis related proteinsecreted frizzled related proteinapoptosisgene expressiongene transfectionearly onset breast cancer[SDGs]SDG3reporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/28723/1/912320B002161.pdf