甘草水提物联合Erastin对二肽基肽酶4诱导结直肠癌细胞铁死亡的作用  

Effect of liquorice extract combined with Erastin on dipeptidyl peptidase-4-induced iron death in colorectal cancer cells

作　　者：陈吉 姚欢欢 尚韬 CHEN Ji;YAO Huan-huan;SHANG Tao(Department of Pharmacy,Huzhou Third People's Hospital,Huzhou 313000,China;Department of Anorectal,The First Affliated Hospital of Zhejiang University of Traditional Chinese Medicine,Zhejiang Provincial Hospital of Traditional Chinese Medicine,Hangzhou 310020,China)

机构地区：[1]湖州市第三人民医院药剂科,湖州313000 [2]浙江中医药大学附属第一医院/浙江省中医院肛肠科,杭州310020

出　　处：《中国新药杂志》2023年第3期270-275,共6页Chinese Journal of New Drugs

基　　金：浙江省中医药科技计划项目(2021ZB095)。

摘　　要：目的:探究甘草水提物(licorice extracts, LE)联合铁死亡诱导剂Erastin能否通过二肽基肽酶4(dipeptidyl peptidase-4,DPP4)诱导人结直肠腺癌上皮细胞(DLD-1)发生铁死亡。方法:将DLD-1细胞随机分为Control组、L-LE组(50μg·mL^(-1)LE)、H-LE组(100μg·mL^(-1)LE)、H-LE+Erastin组(100μg·mL^(-1)LE+30 nmol·L^(-1)Erastin)、H-LE+Erastin+Sita(DDP4抑制剂)组(100μg·mL^(-1)LE+30 nmol·L^(-1)Erastin+31.25μg·mL^(-1)Sita),药物干预24 h。行细胞克隆实验和EDU-594细胞增殖实验测定细胞克隆及增殖情况,ELISA检测细胞Fe2+含量及DPP4活性变化,流式细胞术检测细胞活性氧(reactive oxygen species, ROS)含量,Western blotting检测细胞中溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)、花生四烯酸脂加氢酶3(arachidonate lipoxygenase 3,ALOXE3)蛋白的表达情况。结果:相较于Control组,L-LE,H-LE及H-LE+Erastin处理组细胞的克隆数下降,ROS含量增加(P<0.05或P<0.01);H-LE和H-LE+Erastin处理组细胞的增殖能力降低,Fe^(2+)含量、DPP4活性以及ALOXE3蛋白表达升高,SLC7A11蛋白表达水平降低(P<0.05或P<0.01)。与H-LE+Erastin组相比,加入Sita处理后,细胞的克隆数量、增殖能力及SLC7A11蛋白表达显著增加,而Fe^(2+)含量、DPP4活性、ROS生成量及ALOXE3蛋白表达水平显著降低(P<0.01)。结论:LE联合Erastin可通过DPP4诱导结直肠癌(CRC)细胞发生铁死亡。Objective: To investigate whether licorice extracts(LE) combined with iron death inducer Erastin can induce iron death of colorectal cancer cell DLD1 through dipeptidyl peptidase-4(DPP4). Methods: DLD1 cells were randomly divided into control group, L-LE group(50 μg·mL^(-1)LE), H-LE group(100 μg·mL^(-1)LE), H-LE+Erastin group(100 μg·mL^(-1)LE+30 nmol·L^(-1)Erastin) and H-LE+Erastin+Sita(DPP4 inhibitor) group(100 μg·mL^(-1)LE+30 nmol·L^(-1)Erastin+31.25 μg·mL^(-1)Sita), with drug interventions for 24 h. Cell cloning assay and Edu-594 cell proliferation test were conducted to determine the cloning and proliferation abilities of the cells. ELISA assay was used to detect the Fe2+content and DPP4 activity. Flow cytometry was used to detect reactive oxygen species(ROS) content. Western blotting was used to detect the expression of solute carrier family 7 member 11(SLC7A11) and arachidonate lipoxygenase 3(ALOXE3) proteins. Results: Compared with control group, the cell cloning number decreased while the ROS content increased in L-LE group, H-LE group and H-LE+Erastin group(P<0.05 or P<0.01);and the proliferation ability and Fe^(2+) content decreased, DPP4 activity and ALOXE3 protein expression increased, SLC7A11 protein expression decreased in H-LE and H-LE+Erastin groups(P<0.05 or P<0.01). Compared with H-LE+Erastin group, the addition of Sita significantly increased cell cloning, proliferation and SLC7A11 protein expression, while significantly decreased Fe^(2+) content, DPP4 activity, ROS content and ALOXE3 protein expression(P<0.01). Conclusion: LE combined with Erastin can induce iron death of colorectal cancer(CRC) cell through DPP4.

关 键 词：甘草水提物 Erastin 二肽基肽酶4 结直肠癌细胞 铁死亡 

分 类 号：R965.2[医药卫生—药理学]