奥利司他联合EZH2抑制剂GSK126对胰腺癌细胞脂质代谢重编程的影响  

Effect of Orlistat combined with EZH2 inhibitor GSK126 on lipid metabolism reprogramming in pancreatic cancer cells

作　　者：武晓慧[1] 杨帆 郭新茹 袁轲 马综艺 廉婷 JHA Rajiv Kumar WU Xiaohui;YANG Fan;GUO Xinru;YUAN Ke;MA Zongyi;LIAN Ting;JHA Rajiv Kumar(School of Clinical Medicine,Xi’an Medical University,Xi’an 710021,China;China-Nepal Friendship Medical Research Center of Prof.Rajiv Kumar JHA,Xi’an Medical University,Xi’an 710021,China)

机构地区：[1]西安医学院临床医学院,陕西西安710021 [2]西安医学院中尼友好拉吉姆医学实验室,陕西西安710021

出　　处：《吉林大学学报（医学版）》2023年第1期55-66,共12页Journal of Jilin University：Medicine Edition

基　　金：陕西省卫健委健康科研基金项目(2022D018,2021E021);陕西省西安市未央区科技局科技计划项目(202129);西安医学院博士科研启动基金项目(2020DOC04);西安医学院中尼友好拉吉姆实验室开放基金项目(18LJM02)。

摘　　要：目的:阐明果蝇zeste基因增强子的人类同源物2(EZH2)抑制剂抗胰腺癌疗效不理想的可能机制,为探索新的胰腺癌治疗方案提供依据。方法:采用不同浓度(0~50μmol·L-1)EZH2抑制剂GSK126分别处理胰腺癌PANC-1细胞、CFPAC-1细胞和胰腺转移癌SW1990细胞,CCK-8法检测各组细胞存活率。将3种细胞各分为对照组和GSK126组,GSK126孵育48 h后采用流式细胞术检测各组细胞凋亡率,油红O染色观察各组细胞中脂滴形成情况,检测各组细胞中甘油三酯(TG)水平,实时荧光定量PCR(RT-qPCR)法检测各组细胞中凋亡基因含半胱氨酸的天冬氨酸蛋白水解酶1、3、7、9(Caspase-1、Caspase-3、Caspase-7、Caspase-9)和血管内皮生长因子A(VEGFA)、E-钙黏蛋白(E-cadherin)及干性标志基因CD133、CD24和CD44 mRNA表达水平,以及脂代谢相关基因脂肪酸合成酶(FASN)、乙酰辅酶A羧化酶a(ACACA)、柠檬酸合成酶(CS)、ATP柠檬酸裂解酶(ACLY)、硬脂酰辅酶A去饱和酶(SCD)和酰基辅酶A合成酶短链家族成员2(ACSS2)mRNA表达水平。将3种细胞各分为对照组、GSK126组、FASN抑制剂奥利司他(Orlistat)组和GSK126+Orlistat组,细胞孵育48 h后,CCK-8法检测各组细胞存活率,计算两药联合指数(CI),油红O染色观察各组细胞中脂滴形成情况,检测各组细胞中TG水平,流式细胞术检测各组细胞凋亡率,RT-qPCR法检测各组细胞中凋亡基因Caspase-1、Caspase-3、Caspase-7和Caspase-9及干性基因CD133、CD24和CD44 mRNA表达水平。结果:GSK126呈浓度依赖性抑制胰腺癌细胞增殖。与对照组比较,GSK126组3种细胞凋亡率和凋亡基因mRNA表达水平差异无统计学意义(P>0.05),GSK126组SW1990细胞中VEGFA和PANC-1细胞中E-cadherin mRNA表达水平明显降低(P<0.05),3种细胞中CD133及CFPAC-1和SW1990细胞中CD24 mRNA表达水平均明显升高(P<0.01)。与对照组比较,GSK126组3种细胞中脂滴数量明显增加,TG水平和FASN mRNA表达水平均明显升高(P<0.05或P<0.01),其他Objective:To clarify the possible mechanism of suboptimal efficacy of enhancer of zeste homolog 2(EZH2)inhibitors in pancreatic cancer,and to provide basis for exploring a new treatment plan for pancreatic cancer.Methods:The pancreatic cancer PANC-1 cells,CFPAC-1 cells and pancreatic metastasis cancer SW1990 cells were treated with different concentrations(0-50μmol·L-1)of EZH2 inhibitor GSK126.CCK8 method was used to detect the survival rates of the cells in various groups.Then the cells were divided into control group and GSK126 group.After 48 h of GSK126 incubation,the apoptotic rates of the cells in various groups were detected by flow cytometry,the lipid droplet formation in the cells in various groups were observed with oil red O staining,and the levels of triglyceride(TG)in the cells in various groups were detected.The expression levels of apoptosis genes cysteine-aspartic acid protease(1,3,7 and 9)(Caspase-1,Caspase-3,Caspase-7,Caspase-9),vascular endothelial growth factor A(VEGFA),E-cadherin and stemness markers CD133,CD24 and CD44 mRNA and the expression levels of fat acid synthase(FASN),acetyl CoA carboxylase alpha(ACACA),citrate synthase(CS),ATP citrate lyase(ACLY),stearoyl-CoA desaturase(SCD)and acyl-CoA synthetase short chain family member 2(ACSS2)mRNA were detected by real-time fluorescence quantitative PCR(RT-qPCR)method.The three kinds of cells were divided into control group,GSK126 group,FASN inhibitor Orlistat(Orlistat)group and GSK126+Orlistat group,respectively.After incubated for 48 h,the survival rates of cells in various groups were detected by CCK-8 method,and the combination index(CI)value of the two drugs was calculated.The lipid droplet formation in the cells in various groups were observed with oil red O staining,and the levels of TG in the cells in various groups were detected;the apoptotic rates of cells in various groups were detected by flow cytometry,and the expression levels of Caspase-1,Caspase-3,Caspase-7 and Caspase-9 and CD133,CD24 and CD44 mRNA in the cells in various grou

关 键 词：胰腺肿瘤 化学疗法 EZH2抑制剂 GSK126 脂代谢重编程 脂肪酸合成酶 

分 类 号：R735.9[医药卫生—肿瘤]