缢蛏蛋白组分Y_3的提取及抑癌活性研究

Preliminary studies on extraction and anticancer activity of a protein from Sinonovacula constrzcta

以新鲜缢蛏Sinonovacula constrzcta为材料,经25mmol/LTris-HCl(pH7.5,含0.14mol/LNaCl)缓冲液提取缢蛏粗蛋白、透析、DEAE离子交换层析、SepharoseCL-6B凝胶层析,得到1种具有抑癌生物活性的缢蛏蛋白组分Y3。SDS-PAGE电泳测得该蛋白组分的分子量约为64kD。采用MTT比色法检测Y3对4种肿瘤细胞增殖的影响。结果显示,Y3对Hela和BGC803细胞有明显的抑制作用,而对SKOV3和HepG2抑制作用不明显。此外,探讨了Y3对羟基自由基和超氧阴离子自由基的清除能力。结果显示,该蛋白组分清除自由基的能力很强,在浓度为100μg/ml时,Y3对羟基自由基清除率达到92.71%。在3μg/ml下,对超氧阴离子自由基的清除率达到59.73%。

A protein named as Y3 was purified from Sinonovacula constrzcta （Lamarck） by the method of buffer extraction （pH 7.5,25 mmol/L Tris-HC1 with 0.14 mol/L NaC1）, DEAE ion-exchange chromatography and Sepharose CL-6B chromatography. The molecular weight was determined as 64kD by SDS-PAGE electrophoresis. This protein showed significant inhibitory effect on the growth of Hela and BGC-823. This protein also showed strong scavenging activity on superoxide anion radical and hydroxyl free radical. The scavenging ratio of hydroxyl free rad- ical was up to 92. 71% at 100μg/ml, and that of superoxide anion radical was 59.73% at 3μg/ ml.