- 作者: 王長君; 鄧錦松
- 作者服務機構: 國防醫學院生物及解剖學科; 中央研究院動物研究所; 美國北卡州立大學獸醫學院解剖﹑生理及放射學系
- 中文摘要:
鳥類慕氏抑制素(A-MIS)多株及單株抗體可以確認人類慕氏管(Mullerian duct)來源之女性生殖系統癌細胞株:
Hela S-3, RL 95.2, A-431及NIH: OVCAR-3等,及大白鼠慕氏管上內源(endogenous)哺乳類慕氏抑制素(M-MIS)之
結合業經電子顯微鏡免疫細胞化學技術證實之。標本未經初次抗體處理者;併行於慕氏管側之吳氏管(Wolffian duct);
以及二種非慕氏管來源之癌細胞株,皆呈現MIS負反應。懷孕14天之大白鼠胚慕氏管及培養之癌細胞株,經ABC-MIS-
膠基金(Colloidal gold)及ABC-MIS-過氧化?(Peroxidase)免疫標示後作電子顯微鏡之觀察。本實驗提供初次之電鏡免
疫細胞化學證據,證明:1.A-MIS抗體可以確認M-MIS分子;2.內源之M-MIS結合於慕氏管以及慕氏管來源之癌細胞表
面,亦有結合體-受體(Ligand-receptor)內轉(Internalization)現象;3.因MIS而導致退化之慕氏管,以及生長受抑制
之癌細胞,經本實驗證實為MIS正反應。對 - 英文摘要: Mullerian inhibiting substance (MIS) on rat Mullerian duct (Md), and four Mullerian-derived tumorcells: HeLa S-3, RL-95.2, A-431 and NIH:OVCAR-3 are recognized by the poly- and mono-clonal avian-MIS-antibodies (A-MIS-Abs) using avidin-biotin complex (ABC) immunolabeling techniques. Internali-zation of MIS-ligand complexes was successfully detected in HeLa S-3, OVCAR-3 and RL 95.2 cells. Con-trol groups include: (i) the samples omitted primary antibody treatment, (ii) Wolffian duct by side of Mdin the-genital ridge, and (iii) another two MIS-negative tumor cell lines of non-Mullerian origin: Changhepatoma ascites cell and mouse myeloma cell (X63-Ag 8.653). Genital ridges from rat embryos of 14dof gestation were removed under disection microscope, fixed in 2.5% glutaraldehyde in D-PBS of pH 7.2for 30 min, and sliced into 0.1-0.2 mm thick pieces. A-431, HeLa S-3 and NIH:OVCAR-3 were maintainedin OPTI-MEM culture medium, RL-95.2 was cultured in F12 culture medium. The cells were transferredto 24-well flat bottom culture plates with Thermanox tissue culture coverslips. The immunolabeling offixed and non-fixed samples were processed within the wells. These studies provide first immunocy to-chemical evidences for the similarity between A-MIS and M-MIS molecules by polyclonal and monoclonalA-MIS-Ab. It has also proved that the tumor cell lines, which were subjects of MIS inhibition of cellgrowth, showed MIS binding on cell surfaces.
- 中文關鍵字: Mullerian inhibiting substance; avian; Mullerian duct
- 英文關鍵字: --