拟康氏木霉胞外多糖联合奥沙利铂可体外协同抑制结直肠癌细胞的增殖  

Synergistic effect of polysaccharide from Trichoderma pseudokoningii and oxaliplatin on colorectal cancer cells in vitro

作　　者：李萍[1,2] 袁平川 阙月月 刘筱琴 王国栋[1,2] LI Ping;YUAN Pingchuan;QUE Yueyue;LIU Xiaoqin;WANG Guodong(School of Pharmacy,Wannan Medical College,Wuhu 241002,China;Anhui Provincial Engineering Research Center for Polysaccharide Drugs,Wuhu 241002,China;Chongqing Chemical Industry Vocational College,Chongqing Municipal Vocational and Technical College Application Technology Promotion Center for Key Common Technology in Pharmaceuticals,Chongqing 401220,China)

机构地区：[1]皖南医学院药物研发中心//药学院,安徽芜湖241002 [2]皖南医学院安徽省多糖药物工程技术研究中心//活性生物大分子研究安徽省重点实验室,安徽芜湖241002 [3]重庆化工职业学院//制药领域关键共性工艺重庆市高等职业技术院校应用技术推广中心,重庆401220

出　　处：《南方医科大学学报》2021年第4期504-513,共10页Journal of Southern Medical University

基　　金：国家自然科学基金(81402818);重庆市教育委员会人文社会科学研究项目(17SKG294);安徽省高校自然科学研究项目(KJ2017A257);安徽省大学生创新创业训练项目(S201910368082)。

摘　　要：目的探讨拟康氏木霉胞外多糖(EPS)联合奥沙利铂(Oxa)用药对结肠癌细胞HCT116的协同抑制作用。方法实验分为Control组(0μg/mL)、Oxa组(8μg/mL Oxa)、EPS组(100μg/mL EPS)、EPS+Oxa组(8μg/mL Oxa+100μg/mL EPS)。CCK-8检测细胞活力,CompuSyn软件拟合Fa-CI曲线评价联用效果。流式检测凋亡和周期、划痕实验和transwell实验检测细胞迁移能力,Oxa和EPS相关基因与结直肠癌相关基因取交集进行PPI分析以及GO和KEGG富集分析。结果Oxa单用或与EPS联用处理HCT116细胞,均可使HCT116细胞活力受到抑制,并呈现剂量与时间依耐性,两者联用有明显的协同作用(CI<1)。两药联用组的细胞总凋亡率与Oxa组以及对照组比较均显著升高(P<0.05);联合用药组处于S期的比例高于其他各组。EPS和Oxa均具有抑制HCT116细胞迁移的作用,两者联用后抑制作用更为明显。KEGG分析显示主要涉及耐药、凋亡、血管新生等通路。结论EPS联合奥沙利铂可协同抑制HCT116细胞增殖,促进该细胞凋亡和细胞S周期阻滞、抑制细胞迁移,铂耐药、PI3K-Akt、MAPK等信号通路发挥了关键作用。Objective To explore the synergistic inhibitory effect of polysaccharide from Trichoderma pseudokoningii(EPS)and oxaliplatin(Oxa)on colorectal cancer(CRC)HCT116 cells.Methods HCT116 cells were treated with 8μg/mL Oxa and 100μg/mL EPS alone or in combination,and the changes in cell viability was assessed with CCK-8 assay.CompuSyn software was used for fitting the Fa-CI curve to evaluate the combined effect of the two agents.Flow cytometry was performed to analyze cell apoptosis and cell cycle changes,and wound healing assay and Transwell assay were used to examine the migration ability of the treated cells.Oxa-and EPS-related genes and CRC-related genes were intersected for protein-protein interaction(PPI)analysis and GO and KEGG enrichment analyses.Results Treatment with Oxa alone or in combination with EPS significantly inhibited the viability of HCT116 cells in a dose-and time-dependent manner,and the two agents exhibited a significant synergistic effect(CI<1).The combined treatment with Oxa and EPS resulted in a significantly higher total cell apoptosis rate and a higher percentage of cells in S phase than Oxa alone and the control treatment(P<0.05).EPS and Oxa alone both inhibited the migration of HCT116 cells,and their combination produced a stronger inhibitory effect.GO enrichment analysis of the key genes related with Oxa,EPS and CRC suggested that these genes were involved mainly in such biological processes as exogenous apoptosis signaling,cell response to chemical stress,and reactive oxygen metabolism;KEGG analysis showed that these genes were involved in the pathways of drug resistance,apoptosis and angiogenesis.Conclusion EPS and Oxa can synergistically inhibit the proliferation of HCT116 cells possibly through the PI3K-Akt,MAPK,VEGF,and p53 signaling pathways.

关 键 词：多糖 奥沙利铂 结直肠癌 协同 拟康氏木霉 

分 类 号：R735.34[医药卫生—肿瘤]