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与肿瘤细胞侵袭相关蛋白质的电泳分析。

Electrophoretic analysis of proteins associated with tumor cell invasion.

作者信息

Seftor R E

机构信息

Pediatric Research Institute, Cardinal Glennon Children's Hospital, St. Louis University School of Medicine, MO 63110.

出版信息

Electrophoresis. 1994 Mar-Apr;15(3-4):454-62. doi: 10.1002/elps.1150150162.

Abstract

Polyacrylamide gel electrophoresis is an extremely powerful tool for separating and analyzing protein associated with different diseases and has been invaluable in the identification and analysis of proteins associated with characteristics unique to tumor cells. This study presents data demonstrating the application of conventional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and substrate-incorporated SDS-polyacrylamide gel electrophoresis (zymography) to obtain information about the proteins and catalytically active (or activatable) proteases associated with the process of tumor cell invasion using established human melanoma and breast carcinoma cell lines. Conventional SDS-polyacrylamide gel electrophoresis was used to show that cells sequentially selected from a low invasive human melanoma cell line on the basis of their ability to invade in vitro have an increase and/or addition of six unique proteins on their cell surface. In a different application of SDS-polyacrylamide gel electrophoresis, zymography was used to demonstrate that there is an increase in the levels of gelatinase A in the conditioned medium from three differently invasive human melanoma cell lines coincident with their ability to invade in vitro. Furthermore, the conditioned medium from the most invasive melanoma cell line demonstrated the greatest amount of gelatinase B activity. While the conditioned medium from three human breast carcinoma cell lines contained low levels of both gelatinase A and B, one breast cell line also contained activity associated with stromelysin(s) not seen in the melanoma cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

聚丙烯酰胺凝胶电泳是一种用于分离和分析与不同疾病相关蛋白质的极其强大的工具,在鉴定和分析与肿瘤细胞独特特征相关的蛋白质方面具有重要价值。本研究展示了数据,证明了使用传统的十二烷基硫酸钠(SDS)-聚丙烯酰胺凝胶电泳和掺入底物的SDS-聚丙烯酰胺凝胶电泳(酶谱法),利用已建立的人黑色素瘤和乳腺癌细胞系来获取与肿瘤细胞侵袭过程相关的蛋白质和具有催化活性(或可激活)的蛋白酶的信息。传统的SDS-聚丙烯酰胺凝胶电泳用于显示,基于体外侵袭能力从低侵袭性人黑色素瘤细胞系中依次选择的细胞,其细胞表面有六种独特蛋白质的增加和/或添加。在SDS-聚丙烯酰胺凝胶电泳的另一种应用中,酶谱法用于证明,三种不同侵袭性的人黑色素瘤细胞系的条件培养基中明胶酶A的水平随着它们体外侵袭能力的增强而增加。此外,侵袭性最强的黑色素瘤细胞系的条件培养基显示出最大量的明胶酶B活性。虽然三种人乳腺癌细胞系的条件培养基中明胶酶A和B的水平都很低,但一种乳腺癌细胞系还含有与基质溶解素相关的活性,这在黑色素瘤细胞系中未见。(摘要截取自250字)

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