NDRG1通过ERK通路增强肝细胞癌对索拉菲尼的耐药  

NDRG1 enhances Sorafenib resistance in hepatocellular carcinoma through ERK pathway

作　　者：宋博娇 孙倍成 Song Bojiao;Sun Beicheng(Dept of Hepatobiliary Surgery,Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine,Nanjing 210008;Dept of Hepatobiliary,Pancreatic and Transplantion Surgery,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)

机构地区：[1]南京中医药大学鼓楼临床医学院肝胆外科,南京210008 [2]安徽医科大学第一附属医院肝胆胰及移植外科,合肥230022

出　　处：《安徽医科大学学报》2024年第8期1346-1353,共8页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81930086、82120108012);安徽省高校科研项目(编号:2022AH010070);安徽省临床医学研究转化专项(编号:202204295107020008)。

摘　　要：目的 探究N-myc下游调控基因1(NDRG1)对肝细胞癌(HCC)的作用,以及NDRG1是否影响HCC对索拉菲尼(Sorafenib)的敏感性。方法 通过TCGA数据库预测NDRG1在HCC中的表达水平,并通过蛋白质印迹(WB)实验和免疫组织化学(IHC)染色进行验证。构建NDRG1敲除细胞系进行体外实验,通过肿瘤功能学实验细胞计数试剂盒8(CCK-8)、EdU染色、细胞划痕、Transwell实验研究NDRG1及联合Sorafenib对HCC细胞增殖、迁移与侵袭以及凋亡的影响。利用裸鼠皮下荷瘤进行体内实验,研究NDRG1和Sorafenib对HCC成瘤的影响;通过WB实验和IHC染色确定NDRG1调节HCC对Sorafenib敏感性的通路。结果 WB实验和IHC染色显示,NDRG1在HCC中高表达,与TCGA数据结果一致。肿瘤功能学实验结果表明NDRG1敲除或Sorafenib刺激使HCC细胞增殖、迁移及侵袭能力减弱,肿瘤细胞凋亡增加,而NDRG1敲除联合Sorafenib使HCC细胞增殖、迁移与侵袭能力进一步减弱,肿瘤细胞凋亡进一步增加(P<0.000 1)。小鼠皮下荷瘤模型结果表明NDRG1敲除或Sorafenib刺激使荷瘤体积及质量减小,而NDRG1敲除联合Sorafenib使荷瘤体积及质量进一步减小(P<0.000 1)。WB和IHC结果表明NDRG1敲除联合Sorafenib可降低Erk1/2的磷酸化水平。结论 NDRG1在HCC中高表达,高表达NDRG1促进HCC细胞增殖、迁移和侵袭能力,并抑制肿瘤细胞凋亡。NDRG1通过ERK信号通路增强HCC对Sorafenib的耐药。Objective To investigate the effect of N-myc downstream regulatory gene 1(NDRG1)on Hepatocellular carcinoma(HCC)and whether NDRG1 affects the sensitivity of HCC to Sorafenib.Methods The expression level of NDRG1 in HCC was predicted by TCGA database,and verified by Western blot(WB)and Immunohistochemistry(IHC).NDRG1 knockout cell lines were constructed,followed by cell counting kit-8(CCK-8),EdU,cell scratches and Transwell experiment to investigate the effects of NDRG1 and its combination with Sorafenib on the proliferation,migration and invasion of HCC cells.In addition,flow cytometry was used to detect the apoptosis of HCC cells.The effect of NDRG1 and Sorafenib on HCC tumor formation was studied in vivo by subcutaneous tumor bearing in nude mice.WB and IHC were used to determine the pathway regulating the sensitivity of HCC to Sorafenib.Results WB and IHC confirmed that NDRG1 is highly expressed in HCC,consistent with the results of TCGA data.Tumor functional experiments showed that NDRG1 knockout or Sorafenib stimulation weakened the proliferation,migration,and invasion ability of HCC cells,and increased tumor cell apoptosis.However,NDRG1 knockout combined with Sorafenib further weakened the proliferation,migration and invasion ability of HCC cells,and further increased tumor cell apoptosis(P<0.0001).Mouse subcutaneous tumor model showed that NDRG1 knockout or Sorafenib stimulation reduced the tumor volume and quality,while NDRG1 knockout combined with Sorafenib further reduced the tumor volume and quality(P<0.0001).The WB and IHC results indicated that NDRG1 knockout combined with Sorafenib could reduce the phosphorylation level of Erk1/2.Conclusion NDRG1 is highly expressed in HCC,which promotes the proliferation,migration and invasion of HCC cells,and restricts apoptosis.NDRG1 knockout enhances HCC sensitivity to Sorafenib by reducing ERK signaling pathway phosphorylation.

关 键 词：N-myc下游调控基因1 肝细胞癌 索拉菲尼 ERK通路 

分 类 号：R735.7[医药卫生—肿瘤]