Anagliptin通过SOD-1/ROS调控细胞骨架蛋白生成抑制CT-26细胞迁移的实验研究  

Anagliptin inhibits CT-26 cell migration by regulating cytoskeletal protein production through SOD-1/ROS pathway

作　　者：魏思萌 谭雪[2] 李月 刘逍 武欣 李琦[3] 陈畅 WEI Simeng;TAN Xue;LI Yue;LIU Xiao;WU Xin;LI Qi;CHEN Chang(Department of Pharmacology,School of Pharmacy,Harbin Medical University,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第一医院感染科,150086 [3]哈尔滨医科大学附属肿瘤医院生物治疗中心,150086

出　　处：《临床肿瘤学杂志》2024年第3期209-213,共5页Chinese Clinical Oncology

基　　金：黑龙江省自然科学基金联合引导项目(LH2019H028);国家自然科学基金资助项目(81970382)。

摘　　要：目的研究二肽基肽酶-4(DPP-4)抑制剂Anagliptin对结直肠癌细胞迁移的作用。方法体外培养CT-26细胞;CCK-8实验检测细胞活力;蛋白质印迹实验(Western blotting)检测超氧化物歧化酶-1(SOD-1)和超氧化物歧化酶-2(SOD-2)表达;Transwell小室检测细胞迁移能力;免疫荧光实验检测细胞骨架蛋白(F-actin)和细胞内活性氧原(ROS)形成。结果CCK-8实验显示,2 mmol/L Anagliptin具有促CT-26细胞凋亡作用(P<0.01)。1 mmol/L Anagliptin孵育CT-26细胞后,CT-26细胞迁移能力明显下降(0.375±0.028,P<0.01)。共孵育Anagliptin和Tempol(ROS清除剂)后可明显拮抗由Anagliptin所引起的CT-26细胞迁移抑制(0.527±0.035,P<0.01)及细胞内ROS累积(1.395±0.0553,P<0.01)。CT-26细胞孵育Anagliptin后,细胞内SOD-1表达下降(0.665±0.028,P<0.01),而SOD-2表达无变化(P>0.05)。SOD抑制剂DDC孵育CT-26细胞后,细胞迁移能力明显下降(0.206±0.009,P<0.01)。共孵育Anagliptin和Tempol后拮抗由Anagliptin所引起的CT-26细胞骨架蛋白F-actin合成下降。结论Anagliptin通过SOD-1/ROS途径调控细胞骨架蛋白(F-actin)生成抑制CT-26细胞迁移。Objective To study the effect of dipeptidyl peptidase-4(DPP-4)inhibitor Anagliptin on colorectal cancer cell migration.Methods CT-26 cells were cultured in vitro.CCK-8 assay was used to detect cell viability.The expressions of superoxide dismutase-1(SOD-1)and superoxide dismutase-2(SOD-2)were detected by Western blotting.Cell migration ability was detected in Transwell chamber.Cytoskeleton protein(F-actin)and intracellular reactive oxygen species(ROS)were detected by immunofluorescence assay.Results CCK-8 assay showed that 2 mmol/L Anagliptin could promote apoptosis of CT-26 cells(P<0.01).After CT-26 cells were incubated with 1 mmol/L Anagliptin,the migration ability of CT-26 cells decreased significantly(0.375±0.028,P<0.01).Co-incubation of Anagliptin and Tempol(ROS scavger)could significantly inhibit the migration inhibition of CT-26 cells induced by Anagliptin(0.527±0.035,P<0.01)and intracellular ROS accumulation(1.395±0.0553,P<0.01).After incubating Anagliptin in CT-26 cells,the expression of SOD-1 decreased(0.665±0.028,P<0.01),but the expression of SOD-2 did not change(P>0.05).The migration ability of CT-26 cells was significantly decreased after incubation with SOD inhibitor DDC(0.206±0.009,P<0.01).Co-incubation of Anagliptin and Tempol antagonized the decline in CT-26 cytoskeleton protein F-actin synthesis induced by Anagliptin.Conclusion Anagliptin inhibits CT-26 cell migration by regulating the production of cytoskeleton protein(F-actin)through SOD-1/ROS pathway.

关 键 词：结直肠癌 Anagliptin 细胞迁移 活性氧 细胞骨架 超氧化物歧化酶-1 

分 类 号：R735.3[医药卫生—肿瘤]