摘要
目的探讨瑞芬太尼(RF)调控肝激酶B1(LKB1)/腺苷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)通路对宫颈癌(CC)细胞增殖、迁移及上皮间质转化(EMT)的影响。方法MTT法检测不同浓度RF(0、25、50、100、200、400 ng/mL)对SiHa细胞增殖的影响,将SiHa细胞分为对照(Control)组、顺铂组、RF低剂量(-L)组、RF高剂量(-H)组、RF-H+LKB1抑制剂(Radicicol)组,EdU法检测SiHa细胞增殖,划痕实验和Transwell小室检测SiHa细胞迁移能力,间接免疫荧光法检测上皮间质转化(EMT)相关蛋白表达,Western blot检测LKB1/AMPK/mTOR通路蛋白表达水平。结果与0 ng/mL RF组比,25、50、100、200、400 ng/mL RF处理组SiHa细胞增殖抑制率升高,且呈剂量依赖(P<0.001);与Control组比,RF-L组、RF-H组SiHa细胞EdU阳性率、划痕愈合率、细胞迁移数以及p-mTOR/mTOR比值均降低,LKB1蛋白表达量和p-AMPK/AMPK比值升高(P<0.001),N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)阳性表达减少,E-钙黏蛋白(E-cadherin)阳性表达增加;且RF-H组SiHa细胞中各指标变化幅度高于RF-L组(P<0.05);与RF-H组相比,RF-H+Radicicol组SiHa细胞EdU阳性率、划痕愈合率、细胞迁移数以及p-mTOR/mTOR比值均升高,LKB1蛋白表达量和p-AMPK/AMPK比值降低(P<0.001),且N-cadherin、Vimentin蛋白阳性表达增加,E-cadherin蛋白阳性表达减少。结论RF可能通过调控LKB1/AMPK/mTOR通路抑制CC细胞增殖、迁移及EMT。
Objective To discuss the impacts of remifentanil(RF)on the proliferation,migration,and epithelial mesenchymal transition(EMT)of cervical cancer(CC)cells by regulating the liver kinase B1(LKB1)/AMP-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)pathway.Methods MTT assay was used to detect the effect of different concentrations of RF(0,25,50,100,200,400 ng/mL)on the proliferation of SiHa cells.SiHa cells were assigned into Control group,cisplatin group,low-dose RF(-L)group,high-dose RF(-H)group,and RF-H+LKBl inhibitor(Radicicol)group.EdU method was used to detect SiHa cell proliferation.Scratch experiments and Transwell chambers were used to detect the migration ability of SiHa cells.Indirect immunofluorescence was used to detect the expression of epithelial mesenchymal transition(EMT)-related proteins.Western blot was used to detect the LKB1/AMPK/mTOR pathway proteins.Results Compared with the 0 ng/mL RF group,the proliferation inhibition rate of SiHa cells increased in the 25,50,100,200,and 400 ng/mL RF treatment groups,in a dose-dependent manner(P<0.001).Compared with the Control group,the RF-L and RF-H groups showed a decrease in SiHa cell EdU positivity rate,scratch healing rate,cell migration number,and p-mTOR/mTOR ratio,an increase in LKBl protein and p-AMPK/AMPK ratio(P<0.001),a decrease in N-cadherin and vimentin protein positive expression,and an increase in E-cadherin protein positive expression,and the changes in various indicators in SiHa cells in the RF-H group were higher than those in the RF-L group(P<0.05).Compared with the RF-H group,the RF-H+Radiocicol group showed an increase in SiHa cell EdU positivity rate,scratch healing rate,cell migration number,and p-mTOR/mTOR ratio,a decrease in LKBl protein and p-AMPK/AMPK ratio(P<0.001),an increase in N-cadherin and vimentin protein positive expression,and a decrease in E-cadherin protein positive expression.Conclusion RF may inhibit proliferation,migration,and EMT of CC cells by regulating the LKB1/AMPK/mTOR pathway.
作者
郝亚楠
郝妍妍
石东海
HAO Ya'nan;HAO Yanyan;SHI Donghai(Department of Anesthesiology,Tangshan Maternal and Children Health Hospital,Tangshan,Hebei O63000,China)
出处
《中国优生与遗传杂志》
2025年第10期2147-2152,共6页
Chinese Journal of Birth Health & Heredity
基金
河北省医学科学研究重点课题计划项目(20241254)。
