摘要
探讨山慈菇酯提物(ethyl acetate extract of Cremastra appendiculata,Cr Ap)作用于4T1乳腺癌癌组织中差异蛋白的表达变化,筛选Cr Ap抗4T1乳腺癌的靶点。采用定量蛋白质组学串联质量标签(TMT)标记技术对Cr Ap作用的4T1乳腺癌癌组织进行检测,并筛选4T1组织中发生显著变化的差异蛋白。利用多重数据库对Cr Ap/CON差异蛋白进一步进行生物信息学分析,QPCR检测癌组织中Ly6G、S100a8、S100a9、Tmsb4x和GADPH的mRNA表达量。蛋白质组学共获得15个上调蛋白,1个下调蛋白,其中有4个上调差异蛋白与免疫调节功能密切相关。胸腺素β-4下调,而Toll 4受体结合蛋白、Toll样受体结合蛋白、肥大细胞表达膜蛋白1上调,基因本体论注释分析和功能聚类分析显示这些差异蛋白可能与Cr Ap产生抗乳腺癌作用相关;S100-a8、S100-a9蛋白上调可能与Cr Ap产生促凋亡作用和免疫调节过程相关。此外,KEGG富集通路多与免疫调控过程相关,其中癌症的转录失调通路、IL-17信号通路可能与Cr Ap抗4T1作用和免疫调节机制密切相关。QPCR结果显示Cr Ap能显著提高乳腺癌组织中Ly6G、S100a8、S100a9的表达,而降低Tmsb4x。TMT定量蛋白质组学能有效筛选Cr Ap抗4T1的差异蛋白,其中Cr Ap抗4T1乳腺癌机制和相关的免疫调控作用有望成为新的研究方向。
To explore the different expression of ethyl acetate extract of Cremastra appendiculata(Cr Ap)against 4T_(1)breast cancer for screening its new revelation,tandem mass tag(TMT)-based quantitative proteomics was employed to screen the differential expression proteins treated by Cr Ap in breast cancer tissue.Multiple Bioinformatics databases were used to analyze the differentially expressed proteins.QPCR was used to detect Ly_(6)G,S_(100)a_(8),S_(100)a_(9),Tmsb4x and GADPH mRNA expression in cancer tissues.Totally 16 differentially expressed proteins were identified.Among them,15 proteins were up-regulated and 1 protein was down-regulated.Expression of thymosinβ-4 mRNA was down-regulated significantly,but Toll 4 receptor binding protein,Toll-like receptor binding protein,S100-aS_(100)a_(8)/a_(9) and mast cell expression membrane protein 1 were up-regulated,which may be related to pro-apoptotic effect and immune regulation of Cr Ap against breast cancer.KEGG enrichment pathway analysis of the differentially expressed proteins were enriched to transcriptional misregulation and IL-17 signaling pathway in cancer signaling pathway.Cr Ap significantly increased the expression of Ly_(6)G,S_(100)a_(8),and S_(100)a_(9)genes in breast cancer tissues,and reduced expression of Tmsb4x genes.The immune-related differentially expressed proteins were effectively identified by TMT-based quantitative proteomics between Cr Ap and breast cancer control,and the immunoregulatory effect of Cr Ap against 4T_(1)breast cancer would be expected to become a new research direction.
作者
曹晓东
张楠
杨冬冬
刘颖
王赫
张子英
CAO Xiao-dong;ZHANG Nan;YANG Dong-dong;LIU Ying;WANG He;ZHANG Zi-ying(The Center for New Drug Safety Evaluation and Research of Inner Mongolia Medical University;Public Health of Inner Mongolia Medical University;Basic Medical College of Inner Mongolia Medical University,Hohhot 010110,China;Zhalantun Vocational College,Hulunbeir 162650,China)
出处
《天然产物研究与开发》
CAS
CSCD
2021年第2期246-255,共10页
Natural Product Research and Development
基金
内蒙古自治区自然科学基金(2020MS08136,2020BS08015)
内蒙古自治区高等学校科学研究项目(NJZZ18100)
内蒙古医科大学博士启动金(YKD2018BSJJ005)。
关键词
4T1乳腺癌
蛋白质组学
串联质量标签
山慈菇
4T1 breast cancer
proteomics
tandem repeat sequences
Cremastra appendiculata
