Kobayashi H, Ida W, Fujii T, Terao T, Kawashima Y
Department of Obstetrics and Gynecology, Hamamatsu University School of Medicine.
Nihon Sanka Fujinka Gakkai Zasshi. 1992 May;44(5):571-6.
We purified CA125 antigen from the conditioned media (CM) of eutopic and heterotopic endometrial epithelial cells (EC) as well as ovarian cancer cell lines, SHIN-3 and HOC-I, to determine what molecular weight forms of CA125 antigen were identifiable. Treatment of the high-molecular-weight CA125 antigen with 6M urea yielded a much lower molecular mass peak. After purification by OC125 affinity column chromatography, samples were applied to 3 to 7% polyacrylamide gradient gel and analyzed by Western blot. A single band with a molecular weight (MW) of 200KDa was identified in eutopic EC materials. The CA125 polypeptide of the 110KDa molecule could be detected in all of the CM obtained from heterotopic EC, irrespective of the length of time in the cell culture. A MW of approximately 200 KDa was also observed in some heterotopic EC samples. On the other hand, although the multiple bands with a MW equal to orless than 200KDa were observed in the CM of two ovarian cancer cells, the CA125 polypeptide of 110KDa molecules could not be detected. This preliminary finding offers promise that the 110KDa molecule detection method may be a useful adjunct in the differential diagnosis of heterotopic EC and ovarian cancer.
我们从在位和异位子宫内膜上皮细胞(EC)以及卵巢癌细胞系SHIN-3和HOC-I的条件培养基(CM)中纯化了CA125抗原,以确定可识别的CA125抗原的分子量形式。用6M尿素处理高分子量CA125抗原产生了一个分子量低得多的峰。通过OC125亲和柱色谱纯化后,将样品应用于3%至7%的聚丙烯酰胺梯度凝胶,并通过蛋白质印迹法进行分析。在位EC材料中鉴定出一条分子量(MW)为200KDa的单带。在从异位EC获得的所有CM中均可检测到110KDa分子的CA125多肽,与细胞培养时间长短无关。在一些异位EC样品中也观察到约200KDa的分子量。另一方面,尽管在两种卵巢癌细胞的CM中观察到多条分子量等于或小于200KDa的条带,但未检测到110KDa分子的CA125多肽。这一初步发现表明,110KDa分子检测方法可能是异位EC和卵巢癌鉴别诊断的有用辅助手段。