Kopczyński Z, Kuźniak J, Thielemann A, Kaczmarek J, Rybczyńska M
Oncology, Karol Marcinkowski University of Medical Sciences in Poznań, Poland.
Br J Cancer. 1998 Aug;78(4):466-71. doi: 10.1038/bjc.1998.516.
The aim of our work was quantitative evaluation of the protein and phospholipid fractions of mature erythrocyte membranes separated from women with ovarian cancer. Blood was sampled from 30 women with ovarian cancer, aged 24-79 years, in the third stage of clinical progression of the disease. Phospholipids were separated from membranes by Müller's acidic extraction method and analysed in thin-layer two-dimensional chromatography. On the silica gel plates nine fractions of phospholipids were separated: sphingomyelin (SPH), phosphatidylethanolamine (PE), phosphatidlyserine (PS), phosphatidylcholine (PC), lysophosphatidylcholine (LPC), phosphatidic acid (PA), phosphatidylinositol (Ptd Ins), phosphatidylinositol-4-phosphate (Ptd Ins-4-P), phosphatidylinositol-4,5-diphosphate (Ptd Ins-4,5-P2). The activity of phospholipase C in erythrocyte membranes was determined by Akhrem's spectrophotometric method. Membrane proteins were separated by polyacrylamide gel electrophoresis, SDS-PAGE. It was shown that PS, SPH, LPC and PA fractions were significantly diminished. The concentration of Ptd Ins-4-P and Ptd Ins-4,5-P2 was significantly increased with simultaneous reduction in Ptd Ins level. The inhibition of phospholipase C reached 80%. The quantitative protein evaluation showed a statistically significant decrease in spectrin and a significant increase in 4.1 protein. The quantitative changes, observed in phospholipid and protein fractions, led to the restructuring of the erythrocyte membrane cytoskeleton, which may be connected to increased susceptibility to haemolysis of red blood cells.
我们研究的目的是对从卵巢癌女性患者分离出的成熟红细胞膜中的蛋白质和磷脂组分进行定量评估。从30名年龄在24至79岁、处于疾病临床进展第三阶段的卵巢癌女性患者采集血液。采用米勒酸性提取法从细胞膜中分离出磷脂,并通过薄层二维色谱法进行分析。在硅胶板上分离出九种磷脂组分:鞘磷脂(SPH)、磷脂酰乙醇胺(PE)、磷脂酰丝氨酸(PS)、磷脂酰胆碱(PC)、溶血磷脂酰胆碱(LPC)、磷脂酸(PA)、磷脂酰肌醇(Ptd Ins)、磷脂酰肌醇-4-磷酸(Ptd Ins-4-P)、磷脂酰肌醇-4,5-二磷酸(Ptd Ins-4,5-P2)。采用阿赫雷姆分光光度法测定红细胞膜中磷脂酶C的活性。通过聚丙烯酰胺凝胶电泳(SDS-PAGE)分离膜蛋白。结果显示,PS、SPH、LPC和PA组分显著减少。Ptd Ins-4-P和Ptd Ins-4,5-P2的浓度显著增加,同时Ptd Ins水平降低。磷脂酶C的抑制率达到80%。蛋白质定量评估显示,血影蛋白有统计学意义的减少,而4.1蛋白显著增加。在磷脂和蛋白质组分中观察到的定量变化导致红细胞膜细胞骨架的重构,这可能与红细胞溶血易感性增加有关。