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
魏思萌 1谭雪 2李月 1刘逍 1武欣 1李琦 3陈畅1
作者信息
- 1. 150086 哈尔滨 哈尔滨医科大学药学院药理学教研室
- 2. 150086 哈尔滨医科大学附属第一医院感染科
- 3. 150086 哈尔滨医科大学附属肿瘤医院生物治疗中心
- 折叠
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±0.055 3,P<0.01).CT-26 细胞孵育Ana-gliptin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.
Abstract
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±0.055 3,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-1Key words
Colorectal cancer/Anagliptin/Cell migration/Reactive oxygen species/Cytoskeleton/Superoxide dismutase-1引用本文复制引用
基金项目
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(81970382)
出版年
2024
NETL
NSTL
万方数据