摘要
目的:探究瑞马唑仑(REM)调节鞘氨醇激酶1/鞘氨醇-1-磷酸/鞘氨醇-1-磷酸酯受体3(SPHK1/S1P/S1PR3)信号通路对肝癌细胞增殖、迁移及侵袭的影响。方法:分别用浓度为0、20、40、80、160、320μmol/L的REM处理人肝癌细胞系HepG2,检测细胞存活率,筛选药物浓度;基于细胞存活率结果,将对数期的细胞分为5组:对照组(Control组),瑞马唑仑低、中、高浓度组(REM-L、REM-M、REM-H组,对应20、40和80μmol/L)和瑞马唑仑高浓度+SPHK1激活剂组(REM-H+K6PC-5组)。MTT法检测HepG2细胞存活率;平板克隆实验检测细胞增殖能力,划痕实验检测细胞迁移能力,Transwell小室法检测细胞侵袭能力,流式细胞术检测细胞凋亡情况,Western blot检测细胞中SPHK1、S1P、S1PR3、Bax、Bcl-2蛋白表达水平。结果:与Control组比,REM-L、REM-M、REM-H组HepG2细胞克隆数、划痕愈合率、侵袭数及Bcl-2、SPHK1、S1P、S1PR3、N-cadherin蛋白水平随着REM浓度的升高而逐渐降低,而细胞凋亡率和Bax、E-cadherin蛋白水平则呈现相反趋势(P<0.05);与REM-H组比,REM-H+K6PC-5组HepG2细胞克隆数、划痕愈合率、侵袭数及Bcl-2、N-cadherin、SPHK1、S1P、S1PR3、N-cadherin蛋白水平显著升高,而细胞凋亡率和Bax、E-cadherin蛋白水平则明显下降(P<0.05)。结论:REM可能通过下调SPHK1/S1P/S1PR3信号通路遏制肝癌细胞的生长、转移和侵袭能力,同时诱导细胞凋亡。
Objective:To explore the impacts of remimazolam(REM)on the proliferation,migration,and invasion of hepatocellular carcinoma cells by regulating sphingosine kinase 1/sphingosine 1-phosphate/sphingosine 1-phosphate receptor 3(SPHK1/S1P/S1PR3)signaling pathway.Methods:Human hepatocellular carcinoma cell line HepG2 was treated with REM at concentrations of 0,20,40,80,160,and 320μmol/L respectively.Cell survival rate was measured,and drug concentrations were screened.Based on the results of cell survival rate,the logarithmic phase cells were di-vided into five groups:Control group,low,medium,and high concentrations of remifentanil groups(REM-L,REM-M,REM-H groups;20,40 and 80μmol/L),and high concentrations of remifentanil+SPHK1 activator group(REM-H+K6PC-5 group).MTT assay was used to detect the survival rate of HepG2 cells.Plate cloning experiment was performed to de-tect cell proliferation ability.Scratch experiment was performed to detect cell migration ability.Transwell chamber method was performed to detect cell invasion ability.Flow cytometry was used to detect cell apoptosis.Western blot was used to detect the SPHK1,S1P,S1PR3,Bax,and Bcl-2 proteins in cells.Results:For the Control group,the REM-L,REM-M,and REM-H groups showed that the number of HepG2 cell clones,scratch healing rate,invasion rate,Bcl-2,SPHK1,S1P,S1PR3,and N-cadherin proteins gradually decreased with increasing REM concentration,while the apoptosis rate and Bax,E-cadherin proteins showed an opposite trend(P<0.05).For the REM-H group,the REM-H+K6PC-5 group showed a clear increase in the number of HepG2 cell clones,scratch healing rate,invasion rate,Bcl-2,SPHK1,S1P,S1PR3,and N-cadherin proteins,and a clear decrease in the apoptosis rate and Bax,E-cadherin proteins(P<0.05).Conclusion:REM may inhibit the proliferation,migration,and invasion of hepatocellular carcinoma cells and promote cell apoptosis by suppressing SPHK1/S1P/S1PR3 signaling pathway.
作者
马爱北
李彦儒
齐少霞
孙丙伦
MA Ai-bei;LI Yan-ru;QI Shao-xia;SUN Bing-lun(Department of Anesthesiology,Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine,Cangzhou 160001,China;Department of Western Medicine,Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine,Cangzhou 160001,China;Department of Hepatobiliary and Pancreatic Surgery,Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine,Cangzhou 160001,China)
出处
《中国现代普通外科进展》
2025年第9期680-685,共6页
Chinese Journal of Current Advances in General Surgery
基金
河北省医学科学研究课题计划项目(20220692)。
