人乳腺癌细胞(MCF-7)相关抗原超微结构的定位观察

Ultrastructual Localization of TAA on the MCF-7 Human Breast Cancer Cell

本文以二种免疫电镜法对人乳腺癌细胞系—MCF-7细胞肿瘤相关抗原进行定位观察: 1.直接免疫铁蛋白法。采用戊二醛二步结合法制备铁蛋白偶联人乳腺癌单克隆抗体6C_6B_1B_7A_(12)结合物,并以直接法对MCF-7细胞进行包埋前免疫染色。 2.间接免疫酶标电镜法。以6C_6B_1B_7A_(12)单克隆抗体为第一抗体,过氧化物酶标记兔抗小鼠IgG抗体的结合物为第二抗体对MCF-7细胞进行包埋前免疫染色。 电镜观察结果表明,该肿瘤相关抗原仅分布于MCF-7细胞表面。抗原的存在可能与细胞周期或细胞群体的变异性有关。

Localization of the tumor associative antigen (TAA) on the human breast cancer cell Iine(MCF-7)was performed by two different immuaoelectron microscopic methods.(1) Direct immuno-ferritin method.This is done by taking a two stage method using glutaraldehye as coupling reagent to produce a ferrintin-conjugated monoclonal antibody 6C6B1B7A12 against a membrane antigen on human breast cancer cells, so as to get the result of pre-embed-ding staining for immunocytochemistry of electron microscopy.(2) Indirect immuno-enzyme method.This is done by taking the ascitic monoclonal antibody as the first antibody and the peroxidase conjugated rabbit immunoglobulin to mouse immunoglobulin as the second antibody, so as to achieve the possible result.The result of the immuno-EM examination shows that the TAA identified by monoclonal antibody 6C6B1B7A12 are only distributed on the surface of the MCF-7 cells. It is very possible that the existence of the antigen may be involved in the cell cycle and the variability of cell population.