靶向FoxM1多肽P201对人肝癌HepG2细胞的杀伤作用及关键氨基酸分析  

Killing Effects of FoxM1 Targeted P201 Peptide against Liver Cancer HepG2 Cells and Determination of Key Amino Acids in the Sequence

作　　者：邬怡然 毕振飞 黄家明[1] 崔健[1] 茆灿泉[1] WU Yi-ran BI Zhen-fei HUANG Yia-ming CUI Jian MAO Can-quan(School of Life Science and Engineering, Southwest Jiaotong University, Chengdu, Sichuan, 610031, China)

机构地区：[1]西南交通大学生命科学与工程学院,四川成都610031

出　　处：《现代生物医学进展》2017年第6期1024-1028,1059,共6页Progress in Modern Biomedicine

基　　金：中央高校基本科研业务费专项(SWJTU09ZT28)

摘　　要：目的:初步探究靶向FoxM1多肽P201对人肝癌细胞的杀伤作用、死亡途径以及寻找多肽序列中关键的氨基酸残基。方法:本研究选取人肝癌HepG2细胞为主要研究对象,采用MTT法、AO-EB双染和流式细胞术检测P201多肽对HepG2细胞的杀伤作用和死亡途径,结合模体序列捜索、反义肽氨基酸、虚拟丙氨酸突变和分子对接等方法:确定P201多肽的关键氨基酸。结果:MTT法检测60.0μg/mL P201多肽对HepG2细胞作用48 h抑制率高达96%,形态观察和定量测定显示细胞早期凋亡的发生与P201多肽作用时间和剂量呈一定依赖关系;共确定5个氨基酸(第1、2、4、7、9位残基)为P201多肽的关键氨基酸残基。结论:初步揭示P201多肽对HepG2细胞的强杀伤作用与细胞凋亡相关并确定关键氨基酸为多肽分子的优化和多肽抗癌靶向药物的进一步研发提供了重要的依据与参考。Objective：In this study,we investigated the killing effects and death pathways of FoxM1 targeted dodecapeptide P201 to human liver cancer HepG2 cells as well as key residues in the sequence.Methods：The human liver cancer HepG2 cells was used as a model to elucidate the killing effects and death pathways of P201 peptide by MTT assay,AO-EB staining and flow cytometry.Key residues in P201 sequence were defined by a combination of motif search,antisense peptide theory,virtual alanine mutagenesis and molecular docking.Results：MTT assay revealed that the inhibition rate of HepG2 cells could increase to 96%at the concentration of60.0 μg/mL of P201 peptide after 48 h.AO-EB staining and flow cytometry showed that early apoptosis was in a certain time-and dose-dependent manner to P201 peptide.In addition,five key amino acids（1st,2nd,4th,7th and 9th residues） were defined.Conclusions：The killing effects of P201 peptide to HepG2 cells were strong and related to apoptosis,and key residues were found in P201 sequence.All these results provided important references and insights for the rationality of P201 and further targeted anticancer peptide drug development.

关 键 词：FOXM1 P201多肽 分子对接 癌症 优化 

分 类 号：R965[医药卫生—药理学] R-33[医药卫生—药学]