沉默CD147对前列腺癌LNCaP细胞糖酵解的影响  

Effect of silencing CD147 on glycolysis in prostate cancer LNCaP cells

作　　者：张斯琦 孙美琪 李泽颢 刘丹丹 胡城 方芳 王国庆 ZHANG Siqi;SUN Meiqi;LI Zehao;LIU Dandan;HU Cheng;FANG Fang;WANG Guoqing(Department of Clinical Biochemistry Testing,School of Medical Technology,Beihua University,Jilin 132013,China;Department of Immunology Technology,College of Laboratory Medicine,Jilin Medical University,Jilin 132013,China)

机构地区：[1]北华大学医学技术学院临床生化检验教研室,吉林吉林132013 [2]吉林医药学院检验学院免疫学技术教研室,吉林吉林132013

出　　处：《吉林大学学报（医学版）》2023年第3期634-639,共6页Journal of Jilin University：Medicine Edition

基　　金：吉林省教育厅科学技术研究项目(JJKH20210491KJ);吉林省大学生创新创业训练项目(201913706006);吉林省大学生创新创业训练项目(S202113706039);吉林省卫健委卫生与健康创新项目(2020J076,2020J073);北华大学研究生创新计划项目(2021061)。

摘　　要：目的:探讨CD147对前列腺癌(PCa)LNCaP细胞糖酵解的影响,为PCa临床诊断和治疗提供新的分子标志物和靶点。方法:采用慢病毒感染系统建立沉默CD147的细胞作为LNCaP/shCD147组,同时设立阴性对照组(LNCaP/scramble组)。采用实时荧光定量PCR(RT-qPCR)法检测2组LNCaP细胞中CD147 mRNA表达水平,试剂盒检测2组LNCaP细胞中乳酸(LA)、丙酮酸(PA)和腺嘌呤核苷三磷酸(ATP)浓度及2组LNCaP细胞中丙酮酸激酶(PK)、6-磷酸果糖激酶1(PFK1)和己糖激酶(HK)酶活性,Western blotting法检测2组LNCaP细胞中CD147、HK2、肌肉磷酸果糖激酶(PFKM)和丙酮酸激酶M2(PKM2)蛋白表达水平。结果:与LNCaP/scramble组比较,LNCaP/shCD147组细胞中CD147 mRNA和蛋白表达水平明显降低(P<0.01),表明成功构建沉默CD147的PCa LNCaP细胞模型。与LNCaP/scramble组比较,LNCaP/shCD147组细胞中LA、PA和ATP浓度明显降低(P<0.05或P<0.01)。与LNCaP/scramble组比较,LNCaP/shCD147组细胞中PK、PFK1和HK酶活性明显降低(P<0.05或P<0.01)。Western blotting法检测,与LNCaP/scramble组比较,LNCaP/shCD147组细胞中HK2蛋白表达水平明显降低(P<0.01)。结论:沉默CD147通过调节糖酵解代谢产物、限速酶活性和相关蛋白表达水平抑制LNCaP细胞糖酵解过程。Objective:To discuss the effect of CD147 on the glycolysis of the prostate cancer(PCa)LNCaP cells,and to provide new molecular markers and targets for the clinical diagnosis and treatment of PCa.Methods:The lentivirus infection system was used to establish the cells silencing CD147 as LNCaP/shCD147 group,and the negative control group(LNCaP/scramble group)was set up at the same time.The expression levels of CD147 mRNA in the LNCaP cells in two groups were detected by real-time fluorescent quantitative PCR(RT-qPCR)method;the concentrations of lactic acid(LA),pyruvate(PA),and ATP in the LNCaP cells in two groups were detected by the kits;the activities of pyruvate kinase(PK),6-phosphofructokinase 1(PFK1)and hexokinase(HK)were detected by the kits,and the expression levels of HK2,phosphofructokinase of muscle(PFKM),and pyruvate kinase M2(PKM2)proteins in the LNCaP cells in two groups were detected by Western blotting method.Results:Compared with LNCaP/scramble group,the expression levels of CD147 mRNA and protein in the cells in LNCaP/shCD147 group were significantly decreased(P<0.01);indicating that the PCa LNCaP cell model with silencing CD147 was successfully constructed.Compared with LNCaP/scramble group,the concentrations of LA,PA,and ATP in the cells in LNCaP/shCD147 group were significantly decreased(P<0.05 or P<0.01).Compared with LNCaP/scramble group,the activities of PK,PFK1,and HK enzymes in the cells in LNCaP/shCD147 group were decreased(P<0.05 or P<0.01).The Western blotting results showed that compared with LNCaP/scramble group,the expression level of HK2 protein in the cells in LNCaP/shCD147 group was decreased(P<0.01).Conclusion:Silencing CD147 inhibits the glycolysis of LNCaP cells by regulating the glycolytic metabolites of glycolysis,rate-limiting enzyme activity,and related protein expression levels.

关 键 词：CD147 糖酵解 前列腺肿瘤 LNCAP细胞 己糖激酶2 

分 类 号：R737.25[医药卫生—肿瘤]