人消化系肿瘤染色质非组蛋白电泳图谱分析及免疫原性研究

Study on the PAGE map analysis of various tumor nonhistone proteins and their immunogeneity in human digestive system

从人食管癌、胃癌、直肠癌、肝癌及胎儿肝和从距癌>5cm的癌旁作为对照组织的细胞核中,分别用0.4MNaCl盐提法、苯酚提取法和羟基磷灰石柱层析法提取染色质非组蛋白蛋白质,简称非组蛋白(NHP)进行电泳图谱分析并将不同方法提取的癌NHP作为抗原制备免疫血清做对比免疫组化反应。结果表明①用0.4MNaCl盐提之NHP的质和量更适于作为抗原制备免疫血清和进行免疫组化染色。②以上各种肿瘤NHP与其癌旁NHP之间均有质和量的差异。③不同肿瘤NHP具有组织特异性,然而各种瘤之间也具有相似的共性,即癌NHP之多加带主位于6万d～9万d;多变带区主位于4万d上下处(肝癌NHP之多变带区主位于5万d及8万d);而且肝癌NHP与胎肝NHP有相似处。结果提示癌NHP之多加或加深带可能与癌基因表达的激活有关;而缺如或减弱带可能与载有抗癌基因的染色体区带丢失或基因表达改变有关。

With 3 different techniques, phenol extract, NaCl extract and hydroxyapa-tide chromatography, the nonhistone chromosomal proteins(NHP)were extracted fromhuman esophageal, gastric, rectal and hepatic cancers and the fetal liver,and fromthe tissues(without cancer cells)adjacent to the cancer (5cm in distance) as control.The SDS--polyacrylamide gel electrophoresis plates of the extracted NHP were scan-ned with the CS 930 Scanner.The extracted cancer NHP were used as antigens toprepare the antisera against various tumors,and the immunostaining of NHP wasperformed. The results showed that the NHP extracted with 0.4M NaCl method was superior in quality and quantity to that extracted with the other methods for prepa-ration of the antisera,and that there was difference in quality and quantity of theNHP PAGE bands between various tumor tissues and the adjacent tissues to the can-cer.The results also indicated that various cancer NHP had not only the tissue spe-cificity but also the general character in different kinds of cancers,ie, the additionalbands were located at about 60 kd--90kd, while the positive and negative variablebands were at about 40kd (for the hepatic cancer at both 50 kd and 80 kd). Be-sides, there was similarity in the NHP bands between the hepahc carcinoma andthe fetal liver. The results suggest that the additional and darker bands be related to theactivated oncogene expression, while the disappearance and lighter bands be related tothe chromosomal loss or expression change of the antioncogene.