亚硒酸钠联合阿霉素前体药对胃癌SGC-7901细胞Warburg效应的影响  

Effect of sodium selenite combined with prodrug of adriamycin on the warburg effect of gastric cancer SGC-7901 cells

作　　者：周济 向彩霞 王华桥 杨铁成 丁俊 邹意 樊斌 孙建华 Zhou Ji;Xiang Caixia;Wang Huaqiao;Yang Tiecheng;Ding Jun;Zou Yi;Fan Bin;Sun Jianhua(Department of Gastrointestinal Surgery,the Central Hospital of Enshi Tujia and Miao Autonomous Prefecture,Enshi 445000,China;Hubei Selenium and Human Health Institute,Enshi 445000,China;Department of Gastrointestinal Surgery,Zhongnan Hospital,Wuhan University,the Clinical Medical Research Center of Peritoneal Cancer of Wuhan,Clinical Cancer Study Center of Hubei Provence,Key Laboratory of Tumor Biological Behavior of Hubei Provence,Wuhan 430071,China)

机构地区：[1]恩施州中心医院胃肠外科,445000 [2]恩施州中心医院湖北硒与人体健康研究所,445000 [3]武汉大学中南医院胃肠外科,武汉市腹膜癌临床医学研究中心湖北省肿瘤医学临床研究中心,肿瘤生物学行为湖北省重点实验室,430071

出　　处：《中华实验外科杂志》2020年第3期426-429,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81770283);湖北省自然科学基金(2016CFB668);武汉市腹膜癌临床医学研究中心资助项目(2015060911020462);恩施州科技计划项目(E20170006);恩施州中心医院“砸与人体健康”科研项目。

摘　　要：目的探讨亚硒酸钠联合阿霉素前体药(PADM)对胃癌SGC-7901细胞Warburg效应的影响及其作用机制。方法将SGC-7901(购自美国ATCC公司)细胞分为4组:对照组、亚硒酸钠组(16.94μmol/L)、PADM组(2.00 mg/L)和亚硒酸钠联合PADM组(16.94μmol/L亚硒酸钠+2.00 mg/L PADM)。生化实验检测细胞内葡萄糖摄取量、三磷酸腺苷(ATP)含量及乳酸浓度。反转录-聚合酶链反应(RT-PCR)检测细胞己糖激酶1(HK1)和葡萄糖转运蛋白4(GLUT4)mRNA水平。蛋白质印迹法(Western blot)检测细胞HK1、GLUT4、磷酸化磷脂酰肌醇3-激酶(p-PI3K)和磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)表达水平。多组间比较采用单因素方差分析,均数间两两比较采用SNK检验。结果与对照组比较,亚硒酸钠组、PADM组和亚硒酸钠联合PADM组细胞葡萄糖摄取率[(0.395±0.004)%、(0.467±0.003)%、(0.159±0.006)%]、ATP[(1345.692±18.339)μmol/gprot、(1494.232±32.566)μmol/gprot、(571.075±36.408)μmol/gprot]和乳酸浓度[(7.649±0.348)mmol/gprot、(8.334±0.101)mmol/gprot、(3.188±0.059)mmol/gprot]均明显降低,差异有统计学意义(F=3057.689、252.360、700.295,P<0.05),HK1(mRNA:0.477±0.416、0.573±0.067、0.033±0.006;蛋白:0.492±0.003、0.560±0.003、0.407±0.001)、GLUT4(mRNA:0.326±0.023、0.334±0.017、0.009±0.001;蛋白:0.366±0.002、0.432±0.005、0.205±0.007)、p-PI3K(0.487±0.003、0.589±0.001、0.295±0.006)和p-mTOR(0.432±0.006、0.517±0.005、0.250±0.007)表达均明显下调,差异有统计学意义(F=230.850、382.407、9640.167、3776.600、4791.966、3947.977,P<0.05),且亚硒酸钠联合PADM组比药物单独处理组的抑制效果更明显,差异有统计学意义(t=52.053、72.821、32.912、32.733、21.896、76.347、18.269、13.995、24.001、33.359、42.555、89.685、40.410、48.446、51.192、84.800、35.482、54.547,P<0.05)。结论亚硒酸钠联合PADM可明显抑制SGC-7901细胞的Warburg效应,其作用机制可能与抑制PI3K/mTOR信号通路有关。Objective To investigate the effect of sodium selenite combined with prodrug of adriamycin(PADM)on the warburg effect of gastric cancer SGC-7901 cells(purchased from ATCC,USA)and its mechanism.Methods SGC-7901 cells were divided into 4 groups:control group,sodium selenite group(16.94μmol/L),PADM group(2 mg/L)and sodium selenite+PADM group(16.94μmol/L sodium selenite+2 mg/L PADM).The intracellular glucose uptake,adenosine triphosphate(ATP)content and lactate concentration were determined by biochemical experiments.The mRNA levels of hexokinase 1(HK1)and glucose transporter 4(GLUT4)were detected by reverse transcriptase-polymerase chain reaction(RT-PCR).Western blotting was used to detect the expression of HK1,GLUT4,phosphorylated phosphatidylinositol 3 kinase(p-PI3K)and phosphorylated mammalian target of rapamycin(p-mTOR).Results Compared with the control group,the glucose uptake rate[(0.395±0.004)%,(0.467±0.003)%,(0.159±0.006)%],ATP concentration[(1345.692±18.339)μmol/g prot,(1494.232±32.566)μmol/g prot,(571.075±36.408)μmol/g prot]and lactate concentration[(7.649±0.348)mmol/g prot,(8.334±0.101)mmol/g prot,(3.188±0.059)mmol/g prot]in sodium selenite group,PADM group and sodium selenite+PADM group decreased significantly(F=3057.689,252.360,700.295,P<0.05),while the expression of HK1(mRNA:0.477±0.416,0.573±0.067,0.033±0.006;protein:0.492±0.003,0.560±0.003,0.407±0.001),GLUT4(mRNA:0.326±0.023,0.334±0.017,0.009±0.001;protein:0.366±0.002,0.432±0.005,0.205±0.007),p-PI3K(0.487±0.003,0.589±0.001,0.295±0.006)and p-mTOR(0.432±0.006,0.517±0.005,0.250±0.007)decreased significantly(F=230.850,382.407,9640.167,3776.600,4791.966,3947.977,P<0.05),and the inhibition effect of sodium selenite+PADM group was more obvious than that of drug alone group(F=230.850,382.407,9640.167,3776.600,4791.966,3947.977,P<0.05).Conclusion Sodium selenite combined with PADM can significantly inhibit the warburg effect of SGC-7901 cells,and its mechanism may be related to the inhibition of PI3K/mTOR signaling pathway.

关 键 词：亚硒酸钠 阿霉素前体药 胃癌 Warburg效应 磷脂酰肌醇3-激酶/哺乳动物雷帕霉素靶蛋白信号通路 

分 类 号：R735.2[医药卫生—肿瘤]