摘要
目的:探讨依托咪酯对磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)通路及非小细胞肺癌A549细胞凋亡的影响.方法:设A549细胞组、5-氟尿嘧啶组、依托咪酯低剂量组、高剂量组;5-氟尿嘧啶组、依托咪酯低剂量组、高剂量组分别加入5-氟尿嘧啶组、不同浓度的依托咪酯,使其最终浓度为100.0μg/mL、100.0μg/mL、200.0μg/mL,培养72h.细胞计数试剂盒-8评估细胞增殖水平,流式细胞仪测定细胞凋亡水平,Transwell室测定细胞侵袭性,RT-PCR法及WEST-BLOT法测定细胞磷酸化PI3K(p-PI3K)、磷酸化AKT(p-AKT)mRNA和蛋白水平.结果:与A549细胞组比较,5-氟尿嘧啶组、依托咪酯低、高剂量组OD值、存活率水平、穿膜数、p-PI3K、p-AKT mRNA和蛋白降低(P<0.05),凋亡率水平升高(P<0.05);且随着依托咪酯剂量的增加,依托咪酯各剂量组OD值、存活率、穿膜数、p-PI3K、p-AKT mRNA和蛋白逐渐降低(P<0.05),凋亡率水平逐渐升高(P<0.05).与5-氟尿嘧啶组比较,依托咪酯低剂量组OD值、存活率水平、穿膜数、p-PI3K、p-AKT mRNA和蛋白升高(P<0.05),凋亡率水平降低(P<0.05);依托咪酯高剂量组OD值、存活率水平、穿膜数、p-PI3K、p-AKT mRNA和蛋白降低(P<0.05),凋亡率水平升高(P<0.05).结论:依托咪酯能抑制肺癌A549细胞增殖、侵袭,促进肺癌A549细胞凋亡;其机制与依托咪酯通过减弱p-PI3K、p-AKT mRNA和蛋白磷酸化水平,进而抑制PI3K/AKT通路的激活传导有关.
Objective:To investigate the effect of etomidate on PI3 K/AKT pathway and apoptosis of non-small cell lung cancer A549 cell.Methods:A549 cell group,5-fluorouracil group,etomidate low-dose group and high-dose group were set up.5-fluorouracil group,etomidate low-dose group and high-dose group were added 5-fluorouracil group and etomidate at different concentrations,and the final concentration was100.Oug/mL,100.Oug/mL and 200.Oug/mL for 72 hours.Cell proliferation level was assessed by cell counting kit-8,apoptotic level was measured by flow cytometry,cell invasiveness was measured by Transwell chamber,p-PI3 K,p-AKT mRNA and protein levels were measured by RT-PCR and WEST-BLOT.Results:Compared with A549 cell group,the OD value,survival rate,number of perforation,p-PI3 K,p-AKT gene and protein were significantly decreased in 5-fluorouracil group,low-dose and high-dose etomidate group(P<0.05),apoptotic rate were significantly increased(P<0.05).And with the increase of etomidate dosage,the OD value,survival rate,number of perforation,p-PI3 K,p-AKT gene and protein were gradually decreased(P<0.05),apoptotic rate were gradually increased(P<0.05).Compared with 5-fluorouracil group,the OD value,survival rate,number of perforations,p-PI3 K,p-AKT mRNA and protein in low-dose etomidate group were significantly increased(P<0.05),and apoptotic rate were significantly decreased(P<0.05);while the OD value,survival rate,number of perforation,p-PI3 K,p-AKT mRNA and protein in high-dose etomidate group were significantly decreased(P<0.05),and apoptotic rate were significantly increased(P<0.05).Conclusion:Etomidate can inhibit the proliferation and invasion of lung cancer A549 cells and promote the apoptosis of lung cancer A549 cells.Its mechanism is related to the inhibition of PI3 K/AKT pathway activation and transmission by reducing the levels of p-PI3 K,p-AKT mRNA and protein phosphorylation by etomidate.
作者
杜佳楠
戴勤学
容凤娇
徐夏
DU Jia'nan(Sanya Central Hospital of Hainan Province,Hainan Sanya 572000,China)
出处
《河北医学》
CAS
2020年第7期1066-1071,共6页
Hebei Medicine
基金
海南省卫生计生行业科研项目,(编号:18A200041)。
