旋毛虫肌幼虫ESPs作用于小细胞肺癌H446细胞蛋白组分变化分析  

Changes of protein components in small cell lung cancer H446 cells treated with ESPs of Trichinella spiralis

作　　者：李美辰 潘靖丹 范泽华 陈万楼 陈彩铃 吴合亮 李天歌 孙亚杰 杜娈英[1] LI Mei-chen;PAN Jing-dan;FAN Ze-hua;CHEN Wan-lou;CHEN Cai-ling;WU He-liang;LI Tian-ge;SUN Ya-jie;DU Luan-ying(Department of Pathogen Biology,Chengde Medical College,Chengde 067000,China)

机构地区：[1]承德医学院病原生物学教研室,承德067000

出　　处：《中国人兽共患病学报》2020年第10期821-826,共6页Chinese Journal of Zoonoses

基　　金：河北省教育厅重点项目(No.ZD2014049);河北省高校重点学科冀教高﹝2013﹞4号;校级重点资助项目(No.201709)。

摘　　要：目的分析旋毛虫(Trichinella spiralis)肌幼虫排泄分泌蛋白(excretory-secretory proteins,ESPs)作用于小细胞肺癌H446细胞后蛋白组分的变化。方法制备旋毛虫肌幼虫ESPs,把H446细胞分成实验组和对照组。实验组加入0.85 mg/mL ESPs,对照组加入不含血清的RPMI1640培养基,24 h后收集细胞并提取总蛋白,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离蛋白后酶解,采用液质联用质谱法(LC-MS/MS)对酶解产物进行鉴定,通过Gene Ontology(GO)分析,对鉴定出的实验组与对照组有差异的蛋白质的细胞组分、分子功能和生物过程进行富集分析。结果实验组共鉴定出240种蛋白质,其中有204种为已明确鉴定的蛋白质,22种为未明确鉴定的蛋白质,14种为假定蛋白质。从已知的204种蛋白质中检索出1种与存在于旋毛形肌幼虫排泄分泌蛋白中且与抗肿瘤相关的蛋白质H2A(histone 2A)。对照组鉴定出233种蛋白质。实验组和对照组重合蛋白质191种,非重合蛋白分别有49种和42种。结论旋毛虫肌幼虫ESPs作用于H446细胞后,细胞内蛋白组分发生了复杂变化,提示其抗肿瘤作用机制和过程复杂。This study aimed to analyse changes in the intracellular protein composition of H446 small-cell lung cancer cells after their coculture with excretory-secretory proteins(ESPs)from Trichinella spiralis muscle larvae.The muscle larvae of T.spiralis were collected and used for ESPs preparation.H446 cells in the experimental group were treated with 0.85 mg/mL ESPs,while those in the control group was treated with serum-free RPMI 1640 medium.After 24 h of treatment,the total proteins were collected and extracted.Total proteins were then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and digested by enzymatic hydrolysis.The total proteins were identified by liquid mass spectrometry(LC-MS/MS).Gene Ontology(GO)analysis was used to analyze the enrichment of cellular components,molecular functions and biological processes in proteins which expression in the experimental group differed from that in the control group.The results of LC-MS/MS showed 240 proteins with clear protein bands in the experimental group,of which 204 proteins were identified,22 proteins were unidentified,and 14 proteins were hypothetical.Among the 204 known proteins,one anti-tumor related protein,histone 2A(histone 2A)was found.In the control group,A total of 233 proteins were found and 191 proteins were overlapped between the experimental and control groups.49 and 42 non-overlapping proteins were found in experimental group and control group,respectively.In conclusion,the intracellular protein composition of H446 cells in the experimental group underwent complex changes after culture with ESPs,showing that the anti-tumor mechanism of ESPs and their processes are complex.

关 键 词：旋毛虫 排泄分泌蛋白 LC-MS/MS 质谱分析 

分 类 号：R383[医药卫生—医学寄生虫学]