白花丹醌通过CXCL8/PI3K/AKT糖酵解通路抑制结肠癌细胞增殖、促进凋亡  

Plumbagin inhibits proliferation and promotes apoptosis of colon cancer cells through CXCL8/PI3K/AKT glycolysis pathway

作　　者：杨芳 侯倩倩 李娜 王大庆 陈晓华 YANG Fang;HOU Qianqian;LI Na;WANG Daqing;CHEN Xiaohua(Hengshui People's Hospital,Hebei Hengshui 053000,China;The Fourth Hospital of Hebei Medical University,Hebei Shijiazhuang 050000,China)

机构地区：[1]衡水市人民医院,河北衡水053000 [2]河北医科大学第四医院,河北石家庄050000

出　　处：《现代肿瘤医学》2023年第3期411-416,共6页Journal of Modern Oncology

基　　金：河北省衡水市科技计划(编号:2019014032Z)。

摘　　要：目的:观察白花丹醌对结肠癌细胞Caco-2增殖、凋亡的影响,探究其潜在的作用机制。方法:运用CCK8法、流式细胞术检测不同浓度白花丹醌(4、8、12μmol/L)处理的Caco-2细胞的增殖抑制率、凋亡率。不做任何处理的Caco-2细胞设为Control;脂质体法将si-NC组(转染si-NC)、si-CXCL8组(转染si-CXCL8)转染至Caco-2细胞;8μmol/L的白花丹醌与0.5%DMSO处理的Caco-2细胞设为8μmol/L+DMSO组;8μmol/L的白花丹醌分别与z-VAD-FMK、740Y-P处理的Caco-2细胞设为8μmol/L+z-VAD-FMK组、8μmol/L+740Y-P组。RT-qPCR、Western blot实验检测细胞中CXCL8的mRNA、蛋白表达,CXCL8、M2-型丙酮酸激酶(M2 pyruvate kinase,PKM2)、L-乳酸脱氢酶A(lactate dehydrogenase A,LDHA)、人α-烯醇化酶(apha-enolase,ENO1)、葡萄糖磷酸异构酶(glucose phosphate isomerase,GPI)、磷酸化磷脂酰肌醇3激酶(phosphorylated phosphatidylinositol 3 kinase,p-PI3K)、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-AKT)的蛋白表达。结果:与Control组相比,白花丹醌(4、8、12μmol/L)呈浓度依赖性促进Caco-2细胞增殖抑制率、凋亡率升高,抑制CXCL8的mRNA和蛋白表达。与8μmol/L+DMSO组相比,8μmol/L+z-VAD-FMK组细胞的增殖抑制率、凋亡率明显降低,CXCL8的mRNA和蛋白表达明显升高(P<0.05)。白花丹醌(4、8、12μmol/L)呈浓度依赖性抑制PKM2、LDHA、p-PI3K、p-AKT的蛋白表达。si-CXCL8组PKM2、LDHA、p-PI3K、p-AKT的蛋白表达明显低于si-NC组。740Y-P明显减弱白花丹醌对Caco-2细胞增殖抑制率和凋亡率的促进作用。结论:白花丹醌抑制结肠癌细胞增殖,促进凋亡,其潜在的作用机制与CXCL8/PI3K/AKT糖酵解通路有关。Objective:To observe the effect of plumbagin on cell proliferation and apoptosis of Caco-2,and explore its potential mechanism.Methods:Cell count(CCK8)and flow cytometry were used to measure the proliferation inhibition rate and apoptosis rate of Caco-2 cells treated with plumbagin(4,8,12μmol/L).Caco-2 cells without any treatment were set as the Control group.si-NC group(transfected si-NC)and si-C-X-C motif chemokine ligand 8(CXCL8)group(transfected si-CXCL8)were transfected into Caco-2 cells by liposome method.Caco-2 cells treated with 8μmol/L plumbagin and 0.5%DMSO were set as 8μmol/L+DMSO group.Caco-2 cells treated with 8μmol/L plumbagin and z-VAD-FMK or 740Y-P were set as 8μmol/L+z-VAD-FMK group,8μmol/L+740Y-P group,respectivly.The mRNA and protein expression of CXCL8 were measred by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blot(WB).CXCL8,M2 pyruvate kinase(PKM2),glyceraldehyde-3-phosphate dehydrogenase(LDHA),humanα-enolase(ENO1),glucose phosphate isomerase(GPI),phosphorylated phosphatidylinositol 3 kinase(p-PI3K)and phosphorylated protein kinase B(p-Akt)were measred by Western blot.Results:Compared with the Control group,plumbagin(4,8,12μmol/L)promoted the proliferation inhibition rate and apoptosis rate of Caco-2 cells and inhibited the mRNA and protein expression of CXCL8 in a concentration dependent manner.Compared with 8μmol/L+DMSO group,the proliferation inhibition rate and apoptosis rate significantly decreased,and the mRNA and protein expression of CXCL8 significantly increased in 8μmol/L+z-VAD-FMK group(P<0.05).Plumbagin(4,8,12μmol/L)inhibited the protein expression of PKM2,LDHA,p-PI3K and p-AKT in a concentration dependent manner.The protein expressions of PKM2,LDHA,p-PI3K and p-AKT in si-CXCL8 group were significantly lower than those in si-NC group.740Y-P significantly weakened the promoting effect of plumbagin on the proliferation inhibition rate and apoptosis rate of Caco-2 cells.Conclusion:Plumbagin could inhibit the proliferation and promote

关 键 词：白花丹醌 结肠癌 糖酵解 CXCL8 PI3K/AKT信号通路 

分 类 号：R735.3[医药卫生—肿瘤]