牡蛎活性肽体外诱导HeLa细胞凋亡及其作用机制

食品科学 ›› 2012, Vol. 33 ›› Issue (21): 290-294.

牡蛎活性肽体外诱导HeLa细胞凋亡及其作用机制

余杰，杨振国，陈美珍，钟炼，李伟

汕头大学生物学系

收稿日期:2012-07-04 修回日期:2012-10-22 出版日期:2012-11-15 发布日期:2012-11-09
通讯作者: 陈美珍 E-mail:chenmz@stu.edu.cn
基金资助:
广东省科技计划重点项目;汕头市科技计划项目（2011-156）

Induction and Mechanism of HeLa Cell Apoptosis by Bioactive Peptide from Oyster

Received:2012-07-04 Revised:2012-10-22 Online:2012-11-15 Published:2012-11-09

摘要/Abstract

摘要： 采用复合酶法水解制备牡蛎活性肽，并运用Sephadex G-25柱层析法对酶解液中的肽类进行分离纯化与鉴定；以磺基罗丹明B(SRB)比色法，台盼蓝计数法，Hochest/PI双荧光染色法和DNA Ladder等检测牡蛎活性肽诱导HeLa细胞凋亡的方式，并采用流式细胞术和实时定量PCR分析牡蛎抗活性肽诱导HeLa细胞凋亡的作用机制。经分离纯化得到牡蛎活性肽(bioactive peptide of oyster，BPO)，并测得其分子质量为751D；BPO能有效抑制HeLa细胞增殖，并阻止HeLa细胞G1期向S期的转换，促使凋亡基因表达上调。BPO的作用机制与诱导凋亡基因表达上调和细胞周期阻滞有关。

关键词: HeLa细胞, 牡蛎活性肽, 细胞凋亡, 细胞周期阻滞

Abstract: Flavourzyme and trypsin were used in combination for one-step hydrolysis of oyster homogenate. The resulting hydrolysate was separated by Sephadex G-25 column chromatography. As a result, a bioactive peptide was obtained. The apoptosis inducing effect of the bioactive peptide on HeLa cells was examined by sulforhodamine B colorimetric assay, trypan blue staining assay, Annexin V-FITC/PI double staining assay and DNA ladder assay. Meanwhile, the mechanism was analyzed by flow cytometry and real-time PCR. This peptide had a molecular weight of 751 D. It could effectively inhibit the proliferation of HeLa cells, arrest the cell cycle transition from G1 to S phase and induce the up-regulation of apoptotic gene expression. Its mechanism of action was probably associated with up-regulating apoptotic gene expression and arresting the cell cycle.

Key words: HeLa cell, bioactive peptide from oyster, apoptosis, cell cycle arrest

中图分类号:

Q2544

余?杰，杨振国，陈美珍，钟?炼，李?伟. 牡蛎活性肽体外诱导HeLa细胞凋亡及其作用机制[J]. 食品科学, 2012, 33(21): 290-294.

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