精准医学杂志2025，Vol.40Issue(4)：296-302,7.DOI:10.13362/j.jpmed.202540078
NCOA4对KRAS G12C突变型胰腺癌靶向抑制剂获得性耐药的影响及其机制
Role and mechanism of action of nuclear receptor coactivator 4 in the development of acquired resis-tance to targeted inhibitors in pancreatic cancer with KRAS G12C mutation
摘要
Abstract
Objective To investigate the role and mechanism of action of nuclear receptor coactivator 4(NCOA4)in ac-quired resistance to targeted inhibitors in pancreatic cancer with KRAS G12C mutation.Methods Human pancreatic cancer cell lines CFPAC-1,PANC-1,and MIA-PaCa-2 were cultured in vitro.MIA-PaCa-2 cells were used to establish an adagrasib-resistant cell line(Res-MIA),and a lentivirus was used to establish a drug-resistant MIA-PaCa-2 cell line with NCOA4 knockdown(Res-MIA-KD).Res-MIA cells were divided into groups a and d,while Res-MIA-KD cells were divided into groups b and c;the cells in groups c and d were treated with adagrasib(2 μmol/L),while those in groups a and b received no drug treatment.Subsequently,MIA-PaCa-2 cells were divided into groups A,B,and C,and Res-MIA cells were divided into groups D,E,and F;groups A and D were es-tablished as control groups,the cells in groups B and E were treated with sotorasib(2 μmol/L),and those in groups C and F were treated with adagrasib(2 μmol/L).The transcriptome sequencing technique was used to detect differentially expressed genes be-tween MIA-PaCa-2 and Res-MIA cells;CCK-8 assay was used to assess cell viability in each group;Western blotting was used to measure the relative protein expression levels of NCOA4 and FTH1 in each group;Transwell assay was used to observe cell inva-sion and migration abilities in each group.Male BALB/c nude mice were divided into groups Ⅰ,Ⅱ,Ⅲ,and Ⅳ,and the mice in groups Ⅰ and Ⅳ received orthotopic pancreatic injection of Res-MIA cells,while those in groups Ⅱ and Ⅲ were given orthotopic injection of Res-MIA-KD cells.Since day 14,the mice in groups Ⅲ and Ⅳ were given intraperitoneal injection of adagrasib(10 mg/kg)in normal saline twice a week,while those in groups Ⅰ and Ⅱ were given intraperitoneal injection of an equal volume of normal saline,and tumors were harvested on day 28 to measure tumor volume and weight.Results CCK-8 assay showed that after treatment with adagrasib(0,10,50,100,1 000,or 2 000 nmol/L)for 72 h,there was no significant change in the viability of CFPAC-1 and PANC-1 cells(P＞0.05),while the viability of MIA-PaCa-2 cells decreased significantly with the increase in the con-centration of adagrasib(F=239.24,tLSD=3.68-30.10,P＜0.05).After treatment with sotorasib or adagrasib(0,10,50,100,1 000,or 2 000 nmol/L)for 72 h,there was no significant change in the viability of Res-MIA cells(P＞0.05),and there was no significant difference in cell viability between groups a and d(P＞0.05);groups b and c had a significantly lower cell viability than groups a and d(P＜0.05),and group c had a significantly lower cell viability than group b(P＜0.05).Transcriptome sequencing showed that the expression level of NCOA4 in Res-MIA cells was significantly higher than that in MIA-PaCa-2 cells(P＜0.05).Western blotting showed that groups B and C had a significantly lower relative protein expression level of NCOA4 than group A,and group D had a significantly higher expression level than group A,while group E had a significantly lower expression level than group D(F=366.20,t=4.69-22.71,P＜0.05);groups B and C had a significantly lower relative protein expression level of FTH1 than group A,group D had a significantly higher expression level than group A,and groups E and F had a significantly higher ex-pression level than group D(F=702.16,t=7.36-49.15,P＜0.05);the relative protein expression levels of NCOA4 and FTH1 in Res-MIA-KD cells were significantly lower than those in Res-MIA cells(t=16.27,92.26,P＜0.05).Transwell assay showed that compared with Res-MIA-KD cells,Res-MIA cells had significantly lower numbers of cell migration and invasion(t=9.83,7.19,P＜0.05).Animal experiments showed no significant differences in tumor volume and weight between nude mice of groups Ⅰ andⅣ(P＞0.05),and nude mice of groups Ⅱ and Ⅲ had significantly lower tumor volume and weight than groups Ⅰ and Ⅳ(P＜0.05),while nude mice of group Ⅲ had significantly lower tumor volume and weight than group Ⅱ(P＜0.05).Conclusion Pancreatic cancer with KRAS G12C mutation can enhance its resistance to the targeted agent adagrasib through NCOA4-mediated ferritinophagy,and inhibition of NCOA4 expression can restore the sensitivity of pancreatic cancer cells to adagrasib.关键词
胰腺肿瘤/铁自噬/脱辅铁蛋白质类/核受体共激活因子4/阿达格拉西布/索托拉西布/分子靶向治疗/抗药性,肿瘤/原癌基因蛋白质类p21(ras)/突变Key words
Pancreatic neoplasms/Ferritinophagy/Apoferritins/Nuclear receptor coactivator 4/Adagrasib/Sotorasib/Molecular targeted therapy/Drug resistance,neoplasm/Proto-oncogene proteins p21(ras)/Mutation分类
医药卫生引用本文复制引用
常昊,王亚南,刘世海,王静..NCOA4对KRAS G12C突变型胰腺癌靶向抑制剂获得性耐药的影响及其机制[J].精准医学杂志,2025,40(4):296-302,7.基金项目
青岛市自然科学基金项目(23-2-1-1-189-zyy-d-jch) （23-2-1-1-189-zyy-d-jch）
