| 引用本文:梁桓熙1，王子依1，李晶哲2，马琰岩2，刘长振2，孙震晓1.毛蚶抗癌蛋白主要组分的分离纯化及作用研究[J].世界中医药,2021,(20):. |
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| 毛蚶抗癌蛋白主要组分的分离纯化及作用研究 |
| Study on the Separation,Purification and Function of Anticancer Proteins from Clam Clam |
| 投稿时间：2020-12-23 |
| DOI：10.3969/j.issn.1673-7202.2021.20.009 |
| 中文关键词: 毛蚶抗癌蛋白主要组分C6；层析分离；Colo205；HT-29；A549；K562；细胞凋亡 Caspase |
| English Keywords:Main component of ASAP C6 Chromatographic separation Colo205 HT-29 A549 K562 Apoptosis Caspase |
| 基金项目:国家自然科学基金项目（81473418）；山东省科技发展计划项目（2011YD19007）；财政部资助中国中医科学院基本科研业务费项目（ZZ2018006） |
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| 中文摘要: |
| 目的：对毛蚶抗癌蛋白（ASAP）进行了分离纯化，并研究分离后毛蚶抗癌蛋白主要组分的抗癌作用。方法：依次利用阴离子交换层析、疏水作用层析、凝胶过滤层析对ASAP进行分离；联合p53基因表达激活及CCK-8细胞活力测定检测分离组分的抗癌活性；CCK-8细胞活力测定法检测毛蚶抗癌蛋白主要组分对人结直肠癌Colo205、HT-29、人肺癌A549、人慢性髓性白血病K562细胞活力的影响；Annexin-V/PI双染结合流式细胞术检测细胞凋亡；Western Blotting法检测细胞凋亡相关蛋白：Mcl-1、Bcl-2、Bax、procaspase-3的表达情况。结果：从ASAP中分离得到的毛蚶抗癌活性蛋白主要组分C6在24~72 h内对Colo205、HT-29、A549、K562的细胞活力均有不同程度的抑制作用，其中对Colo205细胞的抑制作用最明显，作用72 h时IC50为5.7 μg/mL；C6可以诱导Colo205细胞凋亡且呈一定剂量依赖性；C6可以抑制抗凋亡蛋白Mcl-1和Bcl-2的表达，上调凋亡蛋白Bax的表达，引起凋亡蛋白caspase-3前体procaspase-3下降。结论：毛蚶抗癌蛋白主要组分C6对人多种癌细胞活力具有抑制作用，可诱导人结直肠癌Colo205细胞发生caspase通路依赖的细胞凋亡。 |
| English Summary: |
| In this study,the anticancer protein(ASAP) of the larvae was isolated and purified,and the anticancer effects of the main components of the anticancer protein of the larvae were studied.Methods:We separated ASAP by anion exchange chromatography,hydrophobic interaction chromatography,and gel filtration chromatography; combined p53 gene expression activation and CCK-8 cell viability assay to track the anticancer activity of the separated components; the CCK-8 cell viability assay detected the effects of the main components of the anti-cancer protein of ASAP on the viability of human colorectal cancer Colo205,HT-29,human lung cancer A549,and human chronic myelogenous leukemia K562; Annexin-V/PI double staining combined with flow cytometry was used to detect cell apoptosis.Results:C6,the main component of the anticancer active protein isolated from ASAP,can inhibit the cell viability of Colo205,HT-29,A549,and K562 to varying degrees within 24 to 72 hours.Among them,it has the most inhibitory effect on Colo205 cells.Obviously,the IC50 was 5.7 μg/mL at 72 h; C6 can induce Colo205 cell apoptosis in a dose-dependent manner; C6 can inhibit the expression of anti-apoptotic proteins Mcl-1 and Bcl-2,and up-regulate the apoptotic protein Bax expression and cause the decrease of the apoptotic protein caspase-3 precursor procaspase-3.Conclusion:C6,the main component of the anti-cancer protein of ASAP,has an inhibitory effect on the viability of a variety of human cancer cells,and can induce caspase pathway-dependent apoptosis in human colorectal cancer Colo205 cells. |
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