人工合成多肽cSN50.1对肝癌细胞HepG2恶性行为的影响及其机制  

Effect of synthetic peptide cSN50.1 on the malignant behavior of hepatocellular carcinoma HepG2 cells and its mechanism

作　　者：辛华[1] 单洪超 栾海艳[2] 阮洋[3] 杨鑫妍 XIN Hua;SHAN Hongchao;LUAN Haiyan;RUAN Yang;YANG Xinyan(The First Affiliated Hospital of Jiamusi University,Jiamusi,Heilongjiang 154007,China;School of Basic Medical Sciences,Jiamusi University,Jiamusi,Heilongjiang 154007,China;The Central Hospital of Jiamusi City,Jiamusi,Heilongjiang 154002,China)

机构地区：[1]佳木斯大学附属第一医院,黑龙江佳木斯154007 [2]佳木斯大学基础医学院,黑龙江佳木斯154007 [3]佳木斯中心医院,黑龙江佳木斯154002

出　　处：《临床肝胆病杂志》2023年第10期2366-2374,共9页Journal of Clinical Hepatology

基　　金：黑龙江省卫生厅项目(20210404010187);佳木斯大学青年创新人才培养支持计划项目(JMSUQP2021016);黑龙江省省属高等学校基本科研业务费团队项目(2022-KYYWF-0656)。

摘　　要：目的 探讨c SN50.1对HepG2细胞增殖、迁移、侵袭和集落形成能力的影响及机制。方法 将HepG2细胞分为6组:c SN50.1 0μmol/L、10μmol/L、30μmol/L、50μmol/L、70μmol/L、90μmol/L组,采用CCK-8实验研究不同浓度c SN50.1对HepG2细胞增殖的影响,并计算半数抑制浓度(IC50);将HepG2细胞分为4组:c SN50.1 0μmol/L、10μmol/L、30μmol/L、50μmol/L,采用细胞划痕、Transwell和细胞克隆实验研究不同浓度cSN50.1对HepG2细胞迁移、侵袭和集落形成能力的影响;将HepG2细胞分为3组:Control组、SP600125组(AP-1信号通路抑制剂)和cSN50.1组,研究AP-1信号通路在cSN50.1对肝癌细胞作用中的影响,采用RT-PCR和Western Blot检测CXCL5和TNF-α的表达以及细胞质和细胞核中c-Jun蛋白的表达;将HepG2细胞分为3组:Control组、PDTC组(NF-κB信号通路抑制剂)和cSN50.1组,研究NF-κB信号通路在cSN50.1对肝癌细胞作用中的影响,采用RT-PCR和Western Blot检测CXCL5和TNF-α的表达以及细胞质和细胞核中NF-κB蛋白的表达。多组间比较采用单因素方差分析,进一步两两比较采用SNK-q检验。结果 与0μmol/L相比,10μmol/L组的增殖、迁移、侵袭和集落形成能力无明显变化(P值均>0.05);30μmol/L组的增殖能力无明显变化(P>0.05),迁移、侵袭和集落形成能力均明显降低(P值均<0.05);50μmol/L组的增殖、迁移、侵袭和集落形成能力均明显降低(P值均<0.01);70μmol/L和90μmol/L组的细胞增殖能力均明显降低(P值均<0.01),但细胞存活率低于50%。与Control组相比,SP600125组、PDTC组和c SN50.1组中CXCL5和TNF-α的基因和蛋白表达均明显降低(P值均<0.05)。与Control组相比,SP600125组、PDTC组和c SN50.1组中细胞核蛋白c-Jun和NF-κB表达均明显降低(P值均<0.05),SP600125组和PDTC组中细胞质蛋白c-Jun和NF-κB表达均明显降低(P值均<0.05),c SN50.1组中细胞质蛋白c-Jun和NF-κB表达明显增高(P<0.05)。结论 c SN50.1可以抑制肝癌细胞的恶性行为,可�Objective To investigate the effect of cSN50.1 on the proliferation,migration,invasion,and colony formation of HepG2 cells and its mechanism.Methods HepG2 cells were divided into cSN50.10μmol/L,cSN50.110μmol/L,cSN50.130μmol/L,cSN50.150μmol/L,cSN50.170μmol/L,and cSN50.190μmol/L groups,and CCK-8 assay was used to investigate the effect of different concentrations of cSN50.1 on the proliferation of HepG2 cells and calculate half-maximal inhibitory concentration(IC50).HepG2 cells were divided into cSN50.10μmol/L,cSN50.110μmol/L,cSN50.130μmol/L,and cSN50.150μmol/L groups,and wound healing assay,Transwell assay,and colony-forming assay were used to investigate the effect of different concentrations of cSN50.1 on the migration,invasion,and colony formation of HepG2 cells.HepG2 cells were divided into Control group,SP600125 group(an inhibitor of the AP-1 signaling pathway),and cSN50.1 group to investigate the influence of the AP-1 signaling pathway on the effect of cSN50.1 on hepatocellular carcinoma cells,and RT-PCR and Western Blot were used to measure the expression of CXCL5,TNF-α,and c-Jun protein in cytoplasm and nucleus.HepG2 cells were divided into Control group,PDTC group(an inhibitor of the NF-κB signaling pathway),and cSN50.1 group to investigate the influence of the NF-κB signaling pathway on the effect of cSN50.1 on hepatocellular carcinoma cells,and RT-PCR and Western Blot were used to measure the expression of CXCL5,TNF-α,and NF-κB protein in cytoplasm and nucleus.A one-way analysis of variance was used for comparison between multiple groups,and the SNK-q test was used for further comparison between two groups.Results Compared with the 0μmol/L group,the 10μmol/L group had no significant changes in proliferation,migration,invasion,and colony formation abilities(P>0.05);the 30μmol/L group had no significant change in proliferation ability(P>0.05),but with significant reductions in migration,invasion,and colony formation abilities(P<0.05);the 50μmol/L group had significant reductions in prol

关 键 词：肝肿瘤 cSN50.1 肿瘤坏死因子Α 趋化因子CXCL5 主动转运 细胞核 

分 类 号：R735.7[医药卫生—肿瘤]