汉麻多肽对Hep3B细胞增殖抑制作用及氨基酸序列分析  

Inhibitory Effect of Hemp Peptides on Hep3B Cell Proliferation and Amino Acid Sequence Analysis

作　　者：魏连会[1] 石杰[1] 李国巍[1] 董艳[1] WEI Lianhui;SHI Jie;LI Guowei;DONG Yan(Daqing Branch of the Heilongjiang Academy of Sciences,Daqing,Heilongjiang 163319,China)

机构地区：[1]黑龙江省科学院大庆分院,黑龙江大庆163319

出　　处：《农产品加工》2025年第1期57-61,67,共6页Farm Products Processing

基　　金：黑龙江省科学院院基金项目(KY2023DQ02);黑龙江省省属科研院所科研业务费项目(CZKYF-2024-1-B39)。

摘　　要：采用木瓜蛋白酶和中性蛋白酶复合酶法水解汉麻粕获得汉麻多肽,经膜分离技术得到分子质量小于2 ku的多肽组分。采用MTT法检测汉麻多肽对体外培养的人肝癌Hep3B细胞增殖的抑制作用,利用倒置显微镜及流式细胞仪观察分析其对肝癌Hep3B细胞的形态、细胞凋亡及细胞周期的影响。利用液相色谱质谱技术对抗肝癌Hep3B细胞增殖活性肽组分进行氨基酸序列分析。结果表明,不同质量浓度的汉麻多肽能够抑制人肝癌Hep3B细胞的生长,且抑制率随汉麻多肽质量浓度的增加而升高。随着汉麻多肽质量浓度的增加,贴壁细胞数量逐渐减少,细胞变圆,且部分细胞核出现浓缩。流式细胞术分析结果表明,汉麻多肽主要将Hep3B细胞阻滞在G2/M期,诱导肝癌Hep3B细胞凋亡,进而抑制肝癌细胞增殖。汉麻多肽活性组分中丰度较高的5个峰对应的肽段氨基酸序列为LPSF,NQANQLDQFSR,FEW,GQNDDRNSIIR,DNGRNVFDGELR,汉麻多肽具有抗肝癌细胞增殖作用。In this study,peptides of hemp meal were obtained by enzymatic hydrolysis of papain and neutral protease.Peptides with molecular mass less than 2 ku were obtained by membrane separation technique.The antitumor efficacy and mechanism of the hemp peptide studied by MTT assays using inverted microscope and fluorescence-activated cell sorter(FACS).To analyze the amino acid sequence of proliferative active peptides of Hep3B cells by liquid chromatography mass spectrometry.The results showed that different concentrations of the hemp peptide could suppress the growth of human hepatoma Hep3B cells,and the growth inhibitory rate increased with increasing concentration of hemp peptide.The number of adherent cells decreased and the number of round cells increased with increasing peptide concentration.In addition some cell nuclei were condensed.Flow cytometry showed that the cells treated with the peptides were arrested G2/M phases.The peptides could induce cell apoptosis and consequently inhibit cell proliferation in Hep3B cell.The amino acid sequences corresponding to the five peaks with high abundance in the active component of hemp peptides were LPSF,NQANQLDQFSR,FEW,GQNDDRNSIIR,DNGRNVFDGELR;In conclusion,the hemp peptides have anti-proliferation effect on hepatocellular carcinoma cells.

关 键 词：汉麻多肽 HEP3B细胞 细胞凋亡 细胞周期 

分 类 号：S664.4[农业科学—果树学]