小檗碱和Wnt/β-catenin信号通路抑制剂对胃癌细胞MKN45增殖凋亡及糖代谢影响研究  

Effects of berberine and Wnt/β-catenin signaling pathway inhibitors on proliferation,apoptosis and glucose metabolism of gastric cancer cell line MKN45

作　　者：张世霞 李聚林[2] 冯五金[2] ZHANG Shi-xia;LI Ju-lin;FENG Wu-jin(Shanxi University of Chinese Medicine,Shanxi,Jinzhong 030619,China;Shanxi Provincial Hospital of traditional Chinese medicine,Shanxi Taiyuan 030012,China)

机构地区：[1]山西中医药大学,山西晋中030619 [2]山西省中医院,太原030012

出　　处：《中国中医基础医学杂志》2020年第7期893-897,共5页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：山西省自然科学基金资助项目(2013011054一6)-功能性消化不良证候要素的提取及适应证规律的研究。

摘　　要：目的:探讨小檗碱和Wnt/β-catenin信号通路抑制剂对胃癌细胞MKN45增殖、凋亡和糖代谢的影响。方法:用不同浓度的小檗碱处理MKN45细胞,噻唑蓝(MTT)检测细胞增殖活性并计算半数抑制浓度(IC 50),分别采用24μmol/L的小檗碱(小檗碱组)、Wnt/β-catenin信号通路抑制剂FH535(FH535组)以及24μmol/L的小檗碱和FH535联合作用(联合组)MKN45细胞,MTT检测细胞增殖活性,流式细胞术检测细胞凋亡,试剂盒检测细胞糖代谢指标己糖激酶、丙酮酸激酶相对活性和培养液上清中乳酸相对含量,Western blot法检测细胞中β-连环蛋白(β-catenin)、c-myc、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)水平。结果:与未使用小檗碱作用的MKN45细胞比较,小檗碱可降低MKN45细胞的吸光度值且呈剂量依赖性,IC 50值为(24.38±2.81)μmol/L,因此选用24μmol/L的小檗碱用于后续实验。与对照组比较,小檗碱组、FH535组、联合组细胞A值均显著降低,细胞凋亡率均显著升高,丙酮酸激酶、己糖激酶及乳酸相对含量均显著降低,β-catenin和c-myc蛋白表达水平显著降低,Cleaved Caspase-3蛋白表达水平显著升高,且联合组的各检测指标均较小檗碱组和FH535组变化明显。结论:小檗碱和Wnt/β-catenin信号通路抑制剂均能够抑制MKN45细胞增殖和糖酵解并诱导其凋亡,且二者具有协同作用。Objective:To investigate the effects of berberine and Wnt/β-catenin signaling pathway inhibitors on proliferation,apoptosis and glucose metabolism in gastric cancer cells MKN45.Methods:Gastric cancer cells were treated with different concentrations of berberine.Cell proliferation activity was measured by thiazolyl blue(MTT),and the half inhibitory concentration(IC50)was calculated.MKN45 cells were treated with 24μmol/L berberine(berberine group),Wnt/β-catenin signaling pathway inhibitor FH535(FH535 group)and 24μmol/L berberine and FH535 combination(combination group).MTT was used to detect the proliferative activity of cells.Flow cytometry was used to detect apoptosis.The kits were used to detect the relative activity of hexokinase and pyruvate kinase in the cell,and the relative content of lactic acid in the culture supernatant.Western blot was used to detectβ-catenin,c-myc,and Cleaved Caspase-3 levels in cells.Results:Compared with MKN45 cells which not treated with berberine,sputum reduced the absorbance of MKN45 cells in a dose-dependent manner.The IC50 value was(24.38±2.81)μmol/L,so 24μmol/L berberine was used for the subsequent experiments.Compared with the control group,the A value of the berberine group,the FH535 group and the combined group were significantly lower,and the apoptotic rate was significantly increased,and the relative contents of pyruvate kinase,hexokinase and lactic acid were significantly decreased,and the expression levels ofβ-catenin and c-myc protein were significantly decreased,and the expression level of Cleaved Caspase-3 protein was significantly increased.The detection indexes of the combined group were significantly lower in the sputum group and the FH535 group.Conclusion:Both berberine and Wnt/β-catenin signaling pathway inhibitors can inhibit the proliferation and glycolysis of MKN45 cells and induce apoptosis,and they have synergistic effects.

关 键 词：胃癌 小檗碱 Wnt/β-catenin信号通路 凋亡 糖代谢 

分 类 号：R285.5[医药卫生—中药学]