摘要
目的探讨咪达唑仑(MDZ)对非小细胞肺癌(NSCLC)细胞增殖、凋亡和迁移的影响及其潜在机制。方法体外培养人NSCLC细胞系SK-MES-1和A549,CCK-8法检测不同浓度的MDZ对NSCLC细胞活力的影响,筛选合适的浓度;将SK-MES-1和A549细胞随即分为对照(Con)组、低剂量咪达唑仑(L-MDZ,100μmol/L)组、中剂量咪达唑仑(M-MDZ,200μmol/L组、高剂量咪达唑仑(H-MDZ,400μmol/L)组、M-MDZ+人重组表皮生长因子(EGF,20 ng/mL)组;给予相应的干预后,采用5-乙基-2’-脱氧尿苷(EdU)染色检测细胞增殖,流式细胞术检测细胞凋亡,Transwell实验检测细胞迁移,Western blot检测EGFR/MEK/ERK信号通路相关蛋白表达。结果MDZ可随浓度升高逐渐抑制SK-MES-1和A549细胞的活性,其中100、200和400μmol/L MDZ的细胞活力抑制率接近50%。与Con组比较,L-MDZ组、M-MDZ组和H-MDZ组EdU阳性细胞率均降低,细胞凋亡率逐渐升高,迁移细胞数均减少,pEGFR/EGFR、pMEK/MEK和pERK/ERK均降低(P<0.05)。M-MDZ组和H-MDZ组之间EdU阳性细胞率、迁移细胞数以及pEGFR/EGFR、pMEK/MEK和pERK/ERK差异无统计学意义(P>0.05)。与M-MDZ组比较,MDZ+EGF组SK-MES-1和A549细胞的EdU阳性细胞率和迁移细胞数均显著升高,细胞凋亡率均明显降低(P<0.05)。结论MDZ可有效抑制NSCLC细胞增殖和迁移,并诱导细胞凋亡,其作用机制可能与抑制EGFR/MEK/ERK信号通路有关。
Objective To investigate the effects of midazolam(MDZ)on the proliferation,apoptosis and migration of non-small cell lung cancer(NSCLC)cells and its potential mechanisms.Methods Human NSCLC cell lines SK-MES-1 and A549 were cultured in vitro,and the effects of MDZ at varying concentrations on the viability of NSCLC cells were detected by the cell counting kit-8(CCK-8)assay to screen for the appropriate concentration of MDZ.Then,NSCLC cells were induced with blank control,low-dose(100μmol/L),medium-dose(200μmol/L)and high-dose MDZ(400μmol/L),and medium-dose MDZ+EGF(20 ng/mL).Cell proliferation,apoptosis,and migration were examined by EdU assay,flow cytometry and Transwell assay,respectively.Protein levels of key molecules in the epidermal growth factor receptor(EGFR)/mitogen-activated protein kinase kinase(MEK)/extracellular signal-regulated kinase(ERK)signaling pathways were detected by Western blot.Results MDZ dose-dependently inhibited the viability of SK-MES-1 and A549 cells,yielding a nearly 50%of inhibitory rate at 100,200 and 400μmol/L.Low-dose,medium-dose and high-dose MDZ induction significantly decreased the rate of EdU-positive cells,migratory cell number,pEGFR/EGFR,pMEK/MEK and pERK/ERK,but increased apoptotic rate(P<0.05).There were no significant differences in the rate of EdU-positive cells,migratory cell number,pEGFR/EGFR,pMEK/MEK and pERK/ERK between NSCLC cells induced with medium-dose and high-dose MDZ(P>0.05).In comparison to medium-dose MDZ group,NSCLC cells induced with medium-dose MDZ plus EGF had significantly higher rate of EdU-positive cells and migratory cell number,but lower apoptotic rate(P<0.05).Conclusion Midazolam effectively inhibits the proliferation and migration of NSCLC cells and induces apoptosis by inhibiting the EGFR/MEK/ERK signaling pathway.
作者
马玮玮
张娟娟
尉家森
MA Weiwei;ZHANG Juanjuan;WEI Jiasen(Medical College,Hexi University,Gansu,Zhangye 734000,China;不详)
出处
《河北医药》
2025年第6期896-901,共6页
Hebei Medical Journal
基金
甘肃省科技计划项目(编号:20JR10RA755)。
关键词
咪达唑仑
非小细胞肺癌
EGFR/MEK/ERK信号通路
增殖
迁移
midazolam
non-small cell lung cancer
epidermal growth factor receptor(EGFR)/mitogen-activated protein kinase kinase(MEK)/extracellular signal-regulated kinase(ERK)signaling pathway
proliferation
migration
