穗花杉双黄酮对小鼠神经母细胞瘤细胞增殖及增殖相关蛋白表达的影响  

Effects of amentoflame on proliferation and proliferation-related protein expression of neuroblastoma cells in mice

作　　者：祝萍萍 苑伟 王鹏珍 ZHU Pingping;YUAN Wei;WANG Pengzhen(Department of Neurology,Guangzhou Red Cross Hospital of Jinan University,Guangdong Province,Guangzhou 510220,China;Department of Surgery,Guangzhou Red Cross Hospital of Jinan University,Guangdong Province,Guangzhou 510220,China;Guangzhou Institute of Traumatic Surgery,Guangzhou Red Cross Hospital of Jinan University,Guangdong Province,Guangzhou 510220,China)

机构地区：[1]暨南大学附属广州红十字会医院神经内科,广东广州510220 [2]暨南大学附属广州红十字会医院普外科,广东广州510220 [3]暨南大学附属广州红十字会医院广州市创伤外科研究所,广东广州510220

出　　处：《中国医药导报》2023年第36期16-20,共5页China Medical Herald

基　　金：广东省基础与应用基础研究基金区域联合基金项目(2020A1515110009);广东省医学科研基金项目(A2021335);广东省中医药局科研项目(20222166)。

摘　　要：目的 探究穗花杉双黄酮(AF)对小鼠神经母细胞瘤细胞(Neuro-2a)及增殖相关蛋白表达的影响。方法 复苏、培养Neuro-2a细胞,取传代后的Neuro-2a细胞用于本研究,CCK-8实验分为5组:0、25、50、75、125μmol/L AF组,细胞处理24 h。细胞流式实验检测AF(75μmol/L AF)对Neuro-2a细胞周期中G1期和S期的百分比的影响,使用荧光定量PCR和Western blot法检测AF(75μmol/L AF)对Neuro-2a细胞增殖相关m RNA和蛋白表达的影响。结果 CCK-8检测结果显示,25、50μmol/L AF组对Neuro-2a细胞活性无显著影响(P>0.05),而75μmol/L AF组则显著降低Neuro-2a细胞活性,差异有统计学意义(P<0.05)。与对照组比较,75μmol/L AF组Neuro-2a细胞G1期细胞百分比升高,差异有统计学意义(P<0.05),与对照组比较,75μmol/L AF组Neuro-2a细胞S期细胞百分比降低,差异有统计学意义(P<0.05),但75μmol/L AF组对Neuro-2a细胞G2期细胞分布无明显作用。荧光定量PCR和Western blot结果显示,75μmol/L AF组可抑制Neuro-2a细胞β-catenin、Cyclin D1、Cyclin E和Survivinm RNA和蛋白表达,促进p21和p16 m RNA和蛋白表达(P<0.05)。结论 75μmol/LAF通过抑制β-catenin、Cyclin D1、Cyclin E和Survivin的表达,促进p21和p16等增殖相关蛋白的表达,进而抑制Neuro-2a细胞进一步增殖。Objective To investigate the effects of amentoflame(AF)on proliferation and proliferation-related protein expression of neuroblastoma cells(Neuro-2a)in mice.Methods Neuro-2a cells were resuscitated,cultured,and selected for this study.The CCK-8 assay was divided into five groups:0,25,50,75 and 125μmol/L groups,the cells were treated for 24 h.The effect of AF(75μmol/L AF)on the percentage of G1 phase and S phase in the cell cycle of Neuro-2a was determined by cytometry.The effects of AF(75μmol/L AF group)on the expression of proliferation-related mRNA and proteins in Neuro-2a cells were detected by fluorescence quantitative PCR and Western blot.Results The results of CCK-8 showed that 25 and 50μmol/L AF group had no significant effect on the activity of Neuro-2a cells(P>0.05),while 75μmol/L group AF significantly decreased the activity of Neuro-2a cells,and the difference was statistically significant(P<0.05).Compared with the control group,the percentage of Neu-2A cells in G1 phase in 75μmol/L AF group was significantly increased,and the difference was statistically significant(P<0.05);compared with the control group,the percentage of S-phase cells of Neuro-2a cells in 75μmol/L AF group was significantly decreased,and the difference was statistically significant(P<0.05);however,75μmol/L AF group had no significant effect on the cell distribution in G2 phase of Neuro-2a cells.Quantitative PCR and Western blot results showed that 75μmol/L AF inhibited the expression ofβ-catenin,CyclinD1,CyclinE,and Survivin mRNA and protein in Neuro-2a cells,and promote expression of p21 and p16 mRNA and protein(P<0.05).Conclusion 75μmol/L AF inhibited the proliferation of Neuro-2a cells by inhibiting the expression ofβ-catenin,CyclinD1,CyclinE,and Survivin,promoting the expression of proliferation-related proteins such as p21 and p16.

关 键 词：神经母细胞瘤 穗花杉双黄酮 细胞周期 凋亡抑制蛋白 细胞周期蛋白D1 

分 类 号：R965[医药卫生—药理学]