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我们采用凝胶过滤和密度梯度离心技术从胰腺癌细胞株SW1990细胞匀浆和细胞培养上清中提取了肿瘤粘液,用无水氢氟酸方法去掉糖链后获得粘液核芯肽,经PAGE和SDS-PAGE及免疫印迹分析表明,从细胞匀浆和细胞培养上清提取的粘液为同一成份。分子量为670kD,亚单位为28kD,以二硫链相连,PAS反应阴性。氨基酸成份分析表明其主要的氨基酸成份有苏氨酸、丝氨酸、脯氨酸,除此之外含有一定量的半胱氨酸,与电泳的结果相符合。酚一硫酸法测定中性糖含量表明不含中性糖。 抗原特性分析表明,用ELISA间接法鉴定为MUC1、MUC2和MUC3的混合物。粘液核芯肽用胃蛋白酶和胰酶处理后,其抗原活性明显降低,而用过碘酸处理后,其抗原活性变化不大。加热及硫酸处理对粘液核;心肽的抗原活性也没有大的影响。二巯基乙醇和SDS可抑制粘液核芯肽的抗原活性。
We used gel filtration and density gradient centrifugation to extract tumor mucin from pancreatic cancer cell line SW1990 cell homogenate and cell culture supernatant. After removing the sugar chain with anhydrous hydrofluoric acid method, mucin core peptide was obtained. SDS-PAGE and immunoblot analysis showed that the mucilage extracted from the cell homogenate and cell culture supernatant was the same. The molecular weight was 670 kD, the subunit was 28 kD, connected by disulfide chains, and the PAS reaction was negative. Amino acid composition analysis showed that the main amino acid components are threonine, serine, and valine, in addition to a certain amount of cysteine, consistent with the results of electrophoresis. The determination of neutral sugar content by the phenol-sulfuric acid method showed no neutral sugar content. Analysis of the antigenic properties revealed indirect ELISA methods to identify mixtures of MUC1, MUC2, and MUC3. After the mucin core peptides were treated with pepsin and trypsin, their antigenic activity was significantly reduced, but after the treatment with periodic acid, their antigenic activity did not change much. Heating and sulfuric acid treatment also had no major effect on the mucin core; the antigenic activity of the cardiac peptide. Dimercaptoethanol and SDS inhibit the antigenic activity of mucin core peptides.