- 學位論文
Phospholipid Scramblase 4 在月經增殖期與分泌期子宮內膜腺體細胞之表現
Expression of Phospholipid Scramblase 4 in Human Endometrial Gland Cell during proliferative and Secretory Phase of Menstrual Cycle
摘要
女性在月經開始時，子宮內膜腺體會隨著週期性的變化而有不同的型態呈現，但是關於其它參與此調控機轉的基因或蛋白報告則非常的少。為了探究這方面的議題，本實驗室過去利用雷射顯微擷取儀和DDRT-PCR鑑定出子宮內膜腺體細胞在增殖期與分泌期具有表現差異的基因，而其中之一為Phospholipid scramblase 4（PLSCR4），目前推測此蛋白與血液凝塊的形成、肥大細胞活化以及吞噬細胞辨識apoptotic cell有關。本實驗希望藉由PLSCR4抗體的製備來了解此蛋白在月經週期子宮內膜變化中所扮演的角色與功能，首先利用PCR與鎳離子親和性管柱等實驗方法來製備抗原，再以皮下注射的方式免疫兔子產生抗體，接著利用細胞株以西方墨點法和IHC stain做抗體的鑑定，最後觀察月經週期之增殖早期和分泌早期的子宮內膜腺體細胞、子宮頸鱗狀上皮癌（SCC）、子宮頸重度異型病變（Cervix Severe dysplasis）與大腸癌等組織之PLSCR4蛋白表現情形。觀察結果發現PLSCR4在月經週期增殖早期之子宮內膜腺體細胞表現較分泌期明顯，推測PLSCR4蛋白可能參與了增殖早期子宮內膜的癒合、修復與增生之過程；而在腫瘤細胞之PLSCR4蛋白表現較正常細胞弱，推測可能是PLSCR4蛋白的缺乏或不足，影響了細胞的老化及凋亡而引發疾病產生。
並列摘要
The human endometrium displays characteristic features both structural and functional across the menstrual cycle. During the proliferative phase, the most notable change is observed in the glandular epithelium, also the process of endometrial healing involves vasoconstriction and clot formation. In previous studies we used LCM to captur the glandular cells of proliferative phase and secretory phase and compared the differences of gene expression by DDRT-PCR technology. Phospholipid scramblase 4 (PLSCR4) is one of the identified genes, its putative functions including initiation of fibrin clot formation, activation of mast cells and facilitate the recognition of apoptotic and injured cells by the reticuloendothelial system. We used Western blot and IHC methods to detect PLSCR4 protein, and found it is less expressed in tumor cells than normal tissues in the Colon cancer 、Cervix Squamous cell Carcinoma and Severe dysplasis. These findings suggest that PLSCR4 protein may be involved in the maintainance of cellular homeostasis. But the roles of PLSCR4 in the cyclic changes of endometrial glandular cell are still not clearly understood and need further studies.