基于IL-6/STAT3信号通路探讨山楂酸对小鼠结肠癌CT26细胞增殖、凋亡的影响  

Effect of maslinic acid on the proliferation and apoptosis of mouse colon cancer CT26 cells based on IL-6/STAT3 signaling pathway

作　　者：李智 陈莉[1] LI Zhi;CHEN Li(College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学药学院,福建福州350122

出　　处：《药学研究》2024年第1期15-18,29,共5页Journal of Pharmaceutical Research

基　　金：福建省自然科学基金面上项目(No.2023J01860);国家自然科学基金青年项目(No.81903855)。

摘　　要：目的从IL-6/STAT3信号通路探讨山楂酸(MA)对小鼠结肠癌CT26细胞增殖、凋亡的影响。方法体外培养CT26细胞,设置空白组、10、20、30、35、40μmoL·L^(-1)MA组。干预24 h后,采用CCK-8法检测山楂酸对细胞活力的影响,采用RT-PCR检测IL-6 mRNA表达水平,采用流式细胞仪检测各组细胞凋亡率,采用Western blotting检测各组细胞中STAT3通路相关蛋白的表达水平。设置空白组、IL-6(20 ng·μL^(-1))刺激组、不同浓度MA与IL-6共刺激组,采用Western blotting检测各组细胞p-STAT3、STAT3蛋白的表达水平。设置空白组、MA组、磷酸酶抑制剂(正钒酸钠)组和MA+正钒酸钠组,采用Western blotting检测各组细胞p-STAT3、STAT3蛋白的表达水平。结果与空白组相比,CCK-8结果显示20、30、35、40、45、50μmoL·L^(-1)浓度的山楂酸均能降低CT26细胞活力(P<0.05),IC50=37.32μmoL·L^(-1);RT-PCR结果显示30、35、40μmoL·L^(-1)给药组细胞中IL-6 mRNA水平下降(P<0.05);流式细胞术结果显示30、35、40μmoL·L^(-1)给药组细胞凋亡率增加(P<0.05);Western blotting结果显示35、40μmoL·L^(-1)给药组细胞中STAT3通路相关蛋白p-STAT3、Bcl-2蛋白表达水平下降(P<0.05),MA+正钒酸钠组能恢复MA单独给药组中被降低的p-STAT3蛋白水平(P<0.05)。结论MA可能通过降低IL-6的表达水平,抑制STAT3的磷酸化,从而降低抗凋亡蛋白Bcl-2的表达,促进肿瘤细胞凋亡,抑制其增殖。Objective To investigate the effect of maslinic acid(MA)on the proliferation and apoptosis of mouse colon cancer CT26 cells through IL-6/STAT3 signaling pathway.Methods CT26 cells were cultured in vitro,then treated with 0,10,20,30,35,and 40μmoL·L^(-1) MA.After 24 hours of intervention,the effect MA on cell viability was detected by CCK-8 method,the expression of IL-6 mRNA was detected by RT-PCR,the apoptosis rate was detected by flow cytometry,and the expression levels of STAT3 pathway-related proteins in each group were detected by Western blotting.Control group,IL-6(20 ng·L^(-1))stimulation group and costimulation groups with different concentrations of MA and IL-6 were set up,and the expression levels of p-STAT3 and STAT3 protein in each group were detected by Western blotting.The expression levels of p-STAT3 and STAT3 protein in cells treated with MA,a phosphatase inhibitor(sodium orthovanadate),or MA+Sodium Orthovanadate simultaneously were detected by Western blotting.Results CCK-8 results showed that MA decreased the viability of CT26 cells compared with control group(P<0.05),IC 50=37.32μmoL·L^(-1).RT-PCR results showed that IL-6 mRNA expression levels in 30,35,40μmoL·L^(-1) administration groups were decreased(P<0.05).Flow cytometry analysis showed that the apoptosis rate was increased in 30,35 and 40μmoL·L^(-1) administration groups(P<0.05).Western blotting results showed that the expression levels of p-STAT3 and Bcl-2 protein in 35 and 40μmoL·L^(-1) administration groups were decreased(P<0.05).Sodium Orthovanadate can restore the p-STAT3 protein level that was reduced in the MA alone group(P<0.05).Conclusion MA may reduce the expression level of IL-6 and further inhibit the phosphorylation of STAT3,thereby reducing the expression of anti-apoptotic protein Bcl-2,promoting tumor cells apoptosis and inhibiting their proliferation.

关 键 词：山楂酸 结肠癌CT26细胞 凋亡 IL-6/STAT3信号通路 

分 类 号：R285.5[医药卫生—中药学]