二肽基肽酶-4抑制剂阿拉格列汀抑制结直肠癌肺转移瘤的形成及其机制  

Dipeptidylpeptidase-4 inhibitor anagliptin inhibiting the formation and mechanism of lung metastasis in colorectal cancer

作　　者：李月 魏思萌 武欣 刘逍 李琦[2] 陈畅 LI Yue;WEI Si-meng;WU Xin;LIU Xiao;LI Qi;CHEN Chang(Department of Pharmacology,School of Pharmacy,Harbin Medical University,Harbin 150086,China;Biotherapy Center,Harbin Medical University Cancer Hospital,Harbin 150086,China)

机构地区：[1]哈尔滨医科大学药学院药理学教研室,哈尔滨150086 [2]哈尔滨医科大学附属肿瘤医院生物治疗中心,哈尔滨150086

出　　处：《解剖学报》2024年第5期582-588,共7页Acta Anatomica Sinica

基　　金：国家自然科学基金(81970382)。

摘　　要：目的探讨二肽基肽酶-4(DPP-4)抑制剂阿拉格列汀(anagliptin)抑制结直肠癌肺转移瘤的作用机制。方法24只雄性BALB/c小鼠随机分为对照组、模型组和anagliptin组。对照组不进行任何处理,模型组于小鼠尾静脉一次性注射浓度为1×10^(9)/L的CT-26细胞溶液0.1 ml后构建肺转移瘤模型,anagliptin组构建肺转移瘤模型0 d后腹腔注射anagliptin 20 mg/(kg·d),14 d后处死实验动物。通过HE染色观察小鼠肺部形态结构。通过MTT实验测定CT-26细胞活力,流式细胞术及活死细胞染色检测CT-26细胞凋亡。活性氧试剂盒测定细胞活性氧簇(ROS)生成,Western blotting测定Bcl-2、Bax,cleaved-Caspase-3、超氧化物歧化酶(SOD)-1和SOD-2表达。结果观察到给予anagliptin后可明显抑制模型组肺组织结构的异常改变。Anagliptin浓度为2 mmol/L时抑制CT-26细胞活力,anagliptin具有促CT-26细胞凋亡的作用,CT-26细胞孵育anagliptin后发现细胞内ROS生成增加。蛋白水平检测发现,CT-26细胞孵育anagliptin后Bax和cleaved-Caspase-3表达升高,而Bcl-2表达下降,给予anagliptin后抑制CT-26细胞SOD-1表达,而对SOD-2无影响。结论Anagliptin通过SOD-1/ROS途径促结直肠癌细胞凋亡,抑制肺转移瘤的形成。Objective To investigate the formation and mechanism of dipeptidylpeptidase-4 inhibitor anagliptin inhibiting lung metastasis of colorectal cancer.MethodsTwenty-four male BALB/c mice were randomly divided into normal group,model group,anagliptin group.The control group did not undergo any treatment.The model group was injected with 0.1 ml CT-26 cells of 10°cells/L.once through the tail vein of the mouse to construct a lung metastasis model,while the anagliptin group was injected intraperitoneally 20 mg/(kg·d)O day after constructing a lung metastasis model.The mice were sacrificed after 14 days.HE staining was used to detect the morphological alteration.Determination of CT-26 cell viability through MTT assay and CT-26 cell apoptosis was detected by flow cytometry and live/dead cell staining.Reactive oxygen species(ROS)production was measured by ROS kit.Western blotting was conducted to measure the expression level of Bcl-2,Bax,cleaved-caspase-3,superoxide dismutase(SOD-1)and SOD-2.Results HE staining showed that the administration of anagliptin could significantly inhibit the abnormal changes in the model group.Anagliptin inhibited the viability of CT-26 cells above 2 mmol/L.Anagliptin promoted the apoptosis of CT-26 cells.Incubation of anagliptin in CT-26 cells significant promoted the production of ROS.Incubation of anagliptin stimulated the expressions of Bax and cleaved-Caspase-3,while down-regulated the expression of Bcl-2 in CT-26 cells.Administration of anagliptin decreased the expression of SOD-1,but not SOD-2.Conclusion Anagliptin promotes apoptosis of colorectal cancer cells and inhibits the formation of lung metastatic tumors through the SOD-1/ROS pathway.

关 键 词：阿拉格列汀 肺转移 超氧化物歧化酶1 活性氧簇 结直肠癌 免疫印迹法 小鼠 

分 类 号：R96[医药卫生—药理学]