咪唑安定通过调控miR-4458/NF-κB通路影响结肠癌细胞的增殖、迁移和侵袭  

Midazolam affecting proliferation,migration,and invasion of colon cancer cells by regulating miR-4458/NF-κB pathway

作　　者：刘爱芬[1] 王辉[2] 王晓鹏 Liu Aifen;Wang Hui;Wang Xiaopeng(Department of Anesthesia,Second Hospital,Tianjin Medical University,Tianjin 300211,China;First Department of Surgery,Gong'an Hospital of Tianjin City,Tianjin 300042,China;Department of Anesthesia,Shanxi Bai Qiu'en Hospital,Taiyuan 030032,China)

机构地区：[1]天津医科大学第二医院麻醉科,300211 [2]天津市公安医院外一科,300042 [3]山西白求恩医院麻醉科,太原030032

出　　处：《国际医药卫生导报》2021年第11期1665-1671,共7页International Medicine and Health Guidance News

基　　金：天津市重点研究项目(2017KYSGAY021)。

摘　　要：目的研究咪唑安定对结肠癌细胞的增殖、迁移和侵袭的影响,并探讨其机制是否与调控微小RNA-4458(miR-4458)/核因子κB(nuclear factorκB,NF-κB)通路有关。方法本研究于2019年12月至2020年10月开展实验。用不同浓度咪唑安定处理结肠癌细胞SW620,噻唑蓝法、Transwell实验检测SW620细胞增殖、迁移和侵袭,蛋白质印迹法检测细胞周期素D1(CyclinD1)、基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)、MMP9、磷酸化的NF-κBp65(phosphorylated NF-κB p65,p-p65)和磷酸化的NF-κB抑制蛋白-α(phosphorylated NF-κB inhibitory protein-α,p-IкBα)表达,实时定量聚合酶链反应分析miR-4458表达。转染miR-4458抑制物至SW620细胞,采用上述方法评估抑制miR-4458表达对SW620细胞增殖、迁移和侵袭的影响。结果咪唑安定处理显著降低SW620细胞增殖活力[(0.843±0.05)、(0.611±0.04)、(0.527±0.03)比(1.089±0.07)]、迁移能力[(147±10.49)个、(117±8.06)个、(83±6.13)个比(179±13.07)个]和侵袭能力[(113±8.67)个、(89±4.71)个、(61±3.79)个比(136±9.14)个](均P<0.05),并下调CyclinD1[(0.70±0.05)、(0.51±0.03)、(0.44±0.03)比(0.83±0.07)]、MMP2[(0.73±0.05)、(0.56±0.04)、(0.45±0.03)比(0.91±0.07)]、MMP9[(0.58±0.04)、(0.41±0.03)、(0.34±0.02)比(0.76±0.06)]、p-p65[(0.70±0.05)、(0.57±0.04)、(0.43±0.03)比(0.83±0.07)]和p-IкBα[(0.61±0.04)、(0.48±0.04)、(0.37±0.02)比(0.71±0.05)]蛋白的表达(均P<0.05),上调miR-4458[(1.43±0.10)、(1.89±0.15)、(2.01±0.14)比(1.00±0.08)]表达(均P<0.05)。抑制miR-4458表达显著增加SW620细胞增殖活力[(1.428±0.11)比(1.093±0.08)]、迁移能力[(237±19.76)个比(183±14.55)个]和侵袭能力[(179±14.12)个比(131±8.14)个](均P<0.05),上调CyclinD1[(1.37±0.11)比(0.85±0.06)]、MMP2[(1.58±0.12)比(0.93±0.08)]和MMP9[(1.11±0.09)比(0.77±0.06)]蛋白表达(均P<0.05)。抑制miR-4458表达可减弱咪唑安定对SW620细胞增殖活力[(0.978±0.07)比(0.534±0.04)]、迁移能力[(17Objective To study the effect of midazolam on the proliferation,migration,and invasion of colon cancer cells,and to explore whether its mechanism is related to the regulation of microRNA-4458(miR-4458)/nuclear factorκB(NFκB)pathway.Methods This study was conducted from December,2019 to October,2020.Colon cancer cells SW620 were treated with different concentrations of midazolam.The cell proliferation,migration,and invasion were detected by Methyl Thiazolyl Tetrazolium and Transwell assays.The expressions of cyclin D1,matrix metalloproteinase 2(MMP2),MMP9,phosphorylated NF-κB p65(p-p65),and phosphorylated NF-κB inhibitory proteinα(p-IкBα)were detected by Western blotting.The expression of miR-4458 was analyzed by real-time quantitative polymerase chain reaction.The miR-4458 inhibitor was transfected into the SW620 cells,and the effect of miR-4458 inhibition on the proliferation,migration,and invasion of the SW620 cells was evaluated by the above methods.Results Midazolam treatment significantly reduced the proliferation[(0.843±0.05),(0.611±0.04),and(0.527±0.03)vs.(1.089±0.07)],migration[(147±10.49),(117±8.06),and(83±6.13)vs.(179±13.07)],and invasion[(113±8.67),(89±4.71),and(61±3.79)vs.(136±9.14)]of the SW620 cells(all P<0.05),down-regulated the expressions of CyclinD1[(0.70±0.05),(0.51±0.03),and(0.44±0.03)vs.(0.83±0.07)],MMP2[(0.73±0.05),(0.56±0.04),and(0.45±0.03)vs.(0.91±0.07)],MMP9[(0.58±0.04),(0.41±0.03),and(0.34±0.02)vs.0.76±0.06)],p-p65[(0.70±0.05),(0.57±0.04),and(0.43±0.03)vs.(0.83±0.07)],and p-IкBα[(0.61±0.04),(0.48±0.04),and(0.37±0.02)vs.(0.71±0.05)](all P<0.05),and up-regulated the expression of miR-4458[(1.43±0.10),(1.89±0.15),and(2.01±0.14)vs.(1.00±0.08)](P<0.05).Inhibiting miR-4458 expression increased the SW620 cells'proliferation[(1.428±0.11)vs.(1.093±0.08)],migration[(237±19.76)vs.(183±14.55)],and invasion[(237±19.76)vs.(183±14.55)](all P<0.05),and up-regulated the expressions of Cyclin D1[(1.37±0.11)vs.(0.85±0.06)],MMP2[(1.58±0.12)vs.(0.93±0.08

关 键 词：咪唑安定 结肠癌 miR-4458 NF-ΚB 细胞增殖 迁移 侵袭 

分 类 号：R735.35[医药卫生—肿瘤]