卵白蛋白-油酸复合物对人结肠癌细胞的HAMLET样生长抑制作用研究
HAMLET- like cell- growth- inhibitory effect of ovalbumin- oleic acid complex on colon cancer cell
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摘要: 乳白蛋白与油酸形成的复合物具有癌细胞生长抑制作用(Human alpha-lactalbumin made lethal to tumor cells,HAMLET)。本文制备了卵白蛋白(Ovalbumin,OVA)和油酸(Oleic acid,OA)的复合物(OVA-OA),通过检测细胞生长抑制、细胞膜通透性、细胞生长周期和凋亡的变化,分析了OVA-OA对人结肠癌细胞(Caco-2)的生长抑制作用以及其中OA对生长抑制作用的贡献。结果表明,当OVA-OA中OA的浓度达到182μmol/L时,OVA-OA对Caco-2细胞有显著(p<0.05)生长抑制作用,273μmol/L时生长抑制作用增强,且随着作用时间的延长(13 h)而增强。在182μmol/L时,OA对Caco-2细胞也有生长抑制作用,其剂量效应关系与OVA-OA一致。但当OA浓度为273μmol/L时,其生长抑制作用弱于OVA-OA的。OVA-OA和OA均会促使乳酸脱氢酶(Lactate dehydrogenase,LDH)释放,增大晚期凋亡和坏死的细胞百分比,阻滞细胞周期在G0/G1期和S期。由此可知,OVA-OA对结肠癌细胞具有HAMLET样生长抑制作用,其中OA起主要作用。OVA-OA对人结肠癌细胞的生长抑制作用是通过损伤细胞膜、阻滞细胞生长周期和促进细胞凋亡来实现的。Abstract: A complex of α- lactalbumin and oleic acid has previously been shown to have growth- inhibitory effect on cancer cells( Human alpha- lactalbumin made lethal to tumor cells,HAMLET). Ovalbumin and oleic acid complex was prepared and its cell growth inhibition effect,along with contribution of oleic acid,were all evaluated on Caco-2 cell by using cell- growth inhibition,cellular membrane integrity,cell cycle and apoptosis assays.The result showed that OVA- OA significantly( p < 0.05) inhibited cell growth at 182 μmol / L( OA),and the inhibitory effect was enhanced as the concentration was increased to 273 μmol / L.The inhibitory effect was also increased as incubation time was extended in a time- dependent manner. OA at 182 μmol / L also exhibited remarkable cell- growth-inhibitory effect in a similar pattern to OVA- OA,while its inhibitory effect was lower than that of OVA- OA at the same OA concentration of 273 μmol / L. Both OVA- OA and OA could increase LDH release rate,lead to cycle arrested at G0/ G1 and S phases,and induce apoptosis.In conclusion,the complex of OVA and OA has HAMLETlike cell- growth- inhibitory effect,which could be mainly contributed to OA.The mechanisms were associated with damage of cellular membrane integrity,arrest of cell cycle and induction of cell apoptosis.