摘要
目的通过不同浓度雷帕霉素干预口腔鳞癌细胞系SCC-4的增殖情况,检测雷帕霉素干预后p-AKT、p-STAT3、p-S6K1及免疫因子在SCC-4中的表达,探讨雷帕霉素对口腔鳞癌细胞体外生长的抑制作用及其分子机制。方法采用20、40、80μmol/L雷帕霉素分别干预体外培养的SCC-4,并设含0μmol/L雷帕霉素的培养液作为对照组,作用24h后,通过MTT检测不同浓度雷帕霉素对于SCC-4增殖的影响;使用Western blotting方法检测雷帕霉素对PI3K/AKT/mTOR信号通路相关相关蛋白(AKT,STAT3,S6K1)磷酸化情况变化;通过酶联免疫吸附法检测雷帕霉素对PI3K/AKT/mTOR信号通路下游共刺激因子B7-H1表达的影响。结果MTT结果显示雷帕霉素对于SCC-4细胞的抑制效应呈明显的浓度依赖关系;酶联免疫吸附法结果表明雷帕霉素能够降低PI3K/AKT/mTOR信号通路下游相关共刺激分子(B7-H1)的浓度;蛋白免疫印迹结果则表明在雷帕霉素干预下SCC-4细胞的PI3K/AKT/mTOR信号通路相关蛋白p-AKT,p-STAT3,p-S6K1表达水平明显降低。结论雷帕霉素有效抑制人口腔鳞癌细胞SCC-4的生长增殖,这可能与调控信号通路中分子的活化转导及其下游共刺激因子有关。
Objective To observe the expression of p-AKT,p-STAT3,p-S6K1 and immune factors in SCC-4after the intervention of rapamycin in different concentrations,investigate the inhibitory effect of rapamycin on the growth of oral squamous cell carcinoma in vitro and its molecular mechanism.Methods 20,40 and 80μmol/L rapamycin were used to interfere with SCC-4 cultured in vitro,while culture medium containing 0μmol/L rapamycin was set as control group.After 24h treatment,the effects of different concentrations of rapamycin on SCC-4 proliferation were detected by MTT.The phosphorylation of rapamycin to PI3K/AKT/mTOR signaling pathway related proteins(AKT,STAT3,S6K1)was detected by western blotting.The effect of rapamycin on the expression of downstream costimulatory factor B7-H1 in PI3K/AKT/mTOR signaling pathway was detected by enzyme-linked immunosorbent assay.Results MTT showed that the inhibitory effect of rapamycin on SCC-4 cells was concentration-dependent.Enzyme-linked immunosorbent assay showed that rapamycin could decrease the concentration of related costimulatory molecules(B7-H1)downstream of PI3K/AKT/mTOR signaling pathway.Protein western blotting results showed that the expression levels of PI3K/AKT/mTOR signaling pathway related proteins p-AKT,p-STAT3 and pS6K1 in SCC-4 cells were significantly decreased after the intervention of rapamycin.Conclusion Rapamycin can inhibit the growth and proliferation of human oral squamous cell carcinoma SCC-4,which may be related to the activation transduction of molecules in the regulatory signaling pathway and its downstream costimulatory factors.
作者
焦会杰
JIAO Huijie(The First Afiliated Hospital of Xingtai Medical College,Xingtai 130500)
出处
《现代口腔医学杂志》
CAS
2023年第2期73-77,共5页
Journal of Modern Stomatology
基金
邢台市科技计划自筹经费项目(2021zc187)。
关键词
雷帕霉素
口腔鳞癌
SCC-4
细胞增殖
Rapamycin
Oral squamous cell carcinoma
SCC-4
Cell proliferation
