硒化秦巴硒菇多糖对A549细胞增殖的抑制作用  

Antitumor efficacy of SeAgaricus blazei Murill(SeABM)on A549 cells

作　　者：孟敏[1] 安红霞 俞永智 王霞[1] 雷文娟 孙延庆[1,4] MENG Min;AN Hongxia;YU Yongzhi;WANG Xia;LEI Wenjuan;SUN Yanqing(Pharmacy Department,Gansu Provincial Hospital,Lanzou 730000,China;The First Clinical College,Gansu Traditional Chinese Medicine University,Lanzhou 730000,China;Bone Surgery Department,Gansu Provincial Hospital,Lanzhou 730000,China;Teching Department,Gansu Provincial Hospital,Lanzhou 730000,China)

机构地区：[1]甘肃省人民医院药剂科,甘肃兰州730000 [2]甘肃中医药大学第一临床学院,甘肃兰州730000 [3]甘肃省人民医院骨外科,甘肃兰州730000 [4]甘肃省人民医院教学部,甘肃兰州730000

出　　处：《甘肃农业大学学报》2022年第3期17-24,共8页Journal of Gansu Agricultural University

基　　金：甘肃省科技计划项目(20JR10RA370);甘肃省卫生行业科研计划资助项目(GSWSKY-2019-23)。

摘　　要：【目的】初步探讨硒化秦巴硒菇多糖(Se-ABM)对人非小细胞肺癌细胞(A549)DNA损伤及细胞周期阻滞的分子机制。【方法】体外培养人结肠癌细胞(HT29)、人肝癌细胞(HepG2)、人宫颈癌细胞(HeLa)和非小细胞肺癌细胞(A549),用Se-ABM和拓扑替康处理(Topotecan)A549细胞,分别用CCK-8试验,流式细胞分析和免疫印迹试验(Western-blot,WB)检测Se-ABM对A549细胞的抗肿瘤作用。【结果】Se-ABM能够抑制HT29、HepG2、HeLa和A549细胞的增殖,其中对A549细胞的增殖抑制最为明显。细胞流式试验表明,Se-ABM在G2/M期阻滞A549细胞,并呈浓度依赖性。Western-blot试验结果显示,Se-ABM能够使A549细胞中DNA损伤相关因子毛细血管扩张性共济失调突变基因(ataxia telangiectasia mutated gene,ATM)、毛细血管扩张性共济失调相关基因(ataxia-telangiectasia related gene,ATR)、抑癌基因(p53)、磷酸化组蛋白(γ-H2AX)的表达量明显升高,同时诱导细胞周期阻滞在G2/M期。此外,在A549细胞中,Se-ABM能够调控凋亡相关蛋白的表达,促使促凋亡因子(BAX)、乳腺癌1号基因(BRCA1)、抗凋亡因子(Bcl-2)、半胱氨酸天冬氨酸蛋白酶3(Caspase 3,Cas 3)和半胱氨酸天冬氨酸蛋白酶(Caspase 9,Cas 9)的表达发生明显变化。【结论】Se-ABM在体外能够明显诱导DNA损伤,从而导致细胞凋亡,Se-ABM具有潜在的抑制A549细胞增殖的效果。【Objective】To investigate the molecular mechanism of DNA damage and cell cycle arrest of human nonsmall cell lung cancer cells(A549)by Se-Agaricus blazei Murill(Se-ABM).【Method】Human colon cancer cells(HT29),human hepatoma cells(HepG2),human cervical cancer cells(HeLa)and A549 cells were cultured in vitro,and non-small cell lung cancer cells(A549)were treated with Se-ABM and topotecan,respectively,and treated with CCK-8 experiments,flow cytometry and Western blot(WB)were used to detect the anti-tumor effect of Se-ABM on A549 cells.【Result】Se-ABM inhibited the proliferation of HT29,HepG2,HeLa and A549 cells,among which the proliferation inhibition of A549 cells was the most obvious.Flow cytometry showed that Se-ABM blocked A549 cells in G2/M phase in a concentrationdependent manner.The results of WB experiment showed that Se-ABM made DNA damage-related factors ataxia telangiectasia mutated gene(ATM)and ataxia-telangiectasia related gene ataxia-telangiectasia related gene(ATR)in A549 cells.Tumor suppressor gene(p53),and phosphorylated histone(γ-H2AX)significantly increased,and cell cycle arrested in G2/M phase.In addition,Se-ABM regulated the apoptosisrelated proteins in A549 cells,including pro-apoptotic factor(BAX),BRCA1 gene(BRCA1),anti-apoptotic factor(Bcl-2),caspase 3(Cas 3)and cysteine-aspartic protease 9(Cas 9),which were markedly changed.【Conclusion】Se-ABM could induce DNA damage and induction of cell apoptosis in vitro.Therefore,Se-ABM has a potential effect to inhibit the proliferation of A549 cells.

关 键 词：硒化秦巴硒菇 拓扑替康 非小细胞肺癌细胞 细胞周期 细胞凋亡 DNA损伤 

分 类 号：R979.1[医药卫生—药品]