摘要
目的观察苦瓜蛋白是否具有诱导K562细胞凋亡的生物学活性,探讨苦瓜蛋白对K562细胞Bcl-2及PCNA表达水平的影响及其作用的分子机制。方法以一定浓度的苦瓜蛋白处理K562细胞12~72小时,CCK-8检测其对细胞生长的影响;流式细胞术(AnnexinⅤ)、细胞形态学(光镜及电镜)等方法检测细胞凋亡;同时应用流式细胞术检测Bcl-2及PCNA蛋白表达的变化。结果苦瓜蛋白对K562细胞生长具有明显的抑制作用;流式细胞术及形态学观察(光镜及电镜)证实一定作用浓度的苦瓜蛋白可诱导K562细胞发生明显的细胞凋亡,且随着作用时间的延长细胞凋亡率逐渐升高。苦瓜蛋白处理组K562细胞中Bcl-2及PCNA蛋白的表达分别为0.33%和98.36%,对照组分别为74.03%和97.63%。结论苦瓜蛋白对K562细胞生长具有明显抑制作用,其作用主要通过诱导K562细胞产生细胞凋亡,而不是抑制细胞增殖。苦瓜蛋白诱导K562细胞凋亡过程中,Bcl-2蛋白表达的下调对调控细胞凋亡有重要作用。
Objective:To explore the apoptosis inducing activity of Momordin from Momordica charantia seeds and its effect on the expression of Bcl-2 gene and proliferating cell nuclear antigen( PCNA). Methods: After the treatment of K562 ceils with appropriated concentration Momordin for 12 to 72 hours. CCK-8 test was employed to determine K562 cells growth; FACScan( FITC- Annexin V staining) , Light and electron microscopy were used to detect apoptosis; The expressions of Bcl-2 and PCNA were examined by FACS- can( FITC- Bcl-2 or FITC- PCNA staining). Results: CCK-8 test showed K562 cells growth was significant inhibited by Momordin; markedly apoptosis was detected by cell morphology and FACScan( FITC-Annexin V) in K562 ceils after treatment by appropriated concentration Momordin and increased in a time-dependent manner; The expressions of Bcl-2 and PCNA in Momordin treated K562 cells were 0. 33% and 97. 63% respectively, and in negative Momordin treated K562 cells were 74. 03% and 98.36% respectively. Conclusion: Momordin can significantly inhibit the growth of K562 cell, its function is performed by inducing apoptosis but not by inhibiting proliferation. Bcl-2 down-expression may probably fimction as an important regulator in the process of apoptosis induced by Momordin.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2006年第11期1002-1005,共4页
Chinese Journal of Immunology
基金
浙江省中医药管理局资助课题(2002C080)
广东省教育厅自然科学研究项目(Z02041)