Phospholipidscramblase4在子宮肌瘤表現的探討

目前醫學界對子宮肌瘤的成因與子宮內膜腺體的變化仍未完全知曉，不少文獻提及卵巢分泌的兩種賀爾蒙－雌激素和黃體酮－是維持子宮內膜健康與子宮肌瘤生長的要素，但對參於調控的基因或蛋白有關的文獻則不多。本實驗室先前利用DDRT-PCR分析出子宮內膜腺體細胞在增殖期及分泌期具有表現差異的基因，本篇研究針對其中PLSCR4，它是Phospholipid scramblase family的成員之一，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。而本實驗希望藉由PLSCR4抗體的製備來了解此蛋白在子宮內膜變化與子宮肌瘤中所扮演的角色與功能，於是利用PCR、鎳離子親和性管柱等實驗來製備蛋白，並以此蛋白去免疫兔子得到抗體，再以IHC stain對月經週期之不同時期的子宮內膜組織做分析，觀察結果發現PLSCR4在增殖期表現量比分泌期表現多，於是再利用西方墨點法與IHC stain來觀察子宮肌瘤，發現PLSCR4在正常肌肉層表現量比子宮肌瘤還多，推測PLSCR4與apoptotic有關，而PLSCR4蛋白的不足影響了細胞的凋亡而引發癌症，至於確實作用機轉仍需進一步研究。

Uterine myoma is a common gynecological tumor and affect nearly one fourth of middle –age wome. It is believed that estrogen and progesterone are important for the proliferation /differentiation of both endometrium and myometrium –the tissue that uterine myoma arising from. However, little is known about the molecular mechanism regulate the tumorigenesis of uterine myoma. We have identified genes expressed differentially between proliferative and secretory endometrium by differential display RTPCR (DDRTPCR), and we tried to explore the role of one of these genes , phospholipid scramblase 4 (PLSCR 4) in the endometrium and uterine myoma.We engineered the DNA construct of PLSC 4 fragment and expressed in E. coli. The fusion protein was purified and used to immunize the rabbit. The specificity and sensitivity of the resultant rabbit anti-PLSCR4 serum were confirmed . Immunhistochemistry assay of endometrium using this anti-PLSCR 4 serum were performed but no significant difference between prliferative and secretory endometrium was noted. The uterine myoma tissues were also analyzed by immunochemistry and western blot, and it showed the expression of PLSCR 4 are lower in uterine myomas than the normal myometrium. This finding suggest the PLSC 4 may play roles in cell apoptosis as other members in the phospholipid scramblase family, and the lower than normal expression may be involved in the tumorigenesis.