阻断Hedgehog信号通路对HepG2.2.15细胞增殖的影响  

Effects of blocking Hedgheg signaling pathway on the proliferation of HepG2.2.15 cell

作　　者：沈璇[1] 黄继锦 张雪[1] 范芳[1] 李长福[1] 

机构地区：[1]遵义医学院生化教研室,贵州遵义563099

出　　处：《贵州医药》2015年第2期106-109,共4页Guizhou Medical Journal

基　　金：贵州省社发攻关项目[黔科合SY(2013)3008];遵义市汇川区科技基金资助项目[(2009)16]

摘　　要：目的探讨环耙明阻断Hedgehog信号通路对肝癌细胞HepG2.2.15增殖的影响。方法培养肝癌细胞HepG2.2.15,用5μmol/L、15μmol/L、25μmol/L环耙明处理HepG2.2.15细胞24h、48h、72h,采用MTT检测环耙明对细胞活力的影响;EDU法检测细胞DNA合成情况;Real-timePCR法检测细胞Gli1的表达情况。结果用5μmol/L、15μmol/L、25μmol/L浓度的环耙明处理HepG2.2.15细胞24h、48h、72h后,MTT检测结果显示细胞活力降低,较空白组差异明显(P<0.05);EDU法检测结果显示25μmol/L的环耙明作用于细胞不同时间后,HepG2.2.15细胞的DNA合成率下降,与空白组相比差异有统计学意义(P<0.01);Real-time PCR法实验结果显示5μmol/L、15μmol/L、25μmol/L浓度的环耙明处理组与对照组相比,Gli1表达水平明显降低(P<0.05)。结论不同浓度环耙明能抑制HepG2.2.15细胞的增殖,减少HepG2.2.15细胞的DNA合成率;其作用机制可能与HepG2.2.15细胞中Gli1、mRNA的表达水平下降有关。Objective To investigate the effects of blocking Hedgheg signaling pathway on the proliferation of HepG2 .2 .15 cell .Method HepG2 .2 .15 cells were divide into blank group ,and the low ,median and high concentra‐tion of cyclopamine treatment groups ,which was treated with the concentrations of cyclopamine were 5 μmol/L , 15 μmol/L ,25μmol/L ,respectively .After 24 h ,48 h ,72 h ,the proliferation of HepG2 .2 .15 cell was detected by MTT .The cellular DNA synthesis was detected by EdU .The expression levels of Gli1 ,mRNA were measured by rea1‐time PCR .Result MTT assay results showed that the cyclopamine treatment groups with the concentration of 5 μmol/L ,15 μmol/L ,25 μmol/L was significantly reduced cell proliferation compared with the control group , which there were statistical significantly differences （P〈 0 .05） .EdU detection results showed the cyclopamine treatment groups decreased the cellular DNA synthesis compared with the control group ,individually .Which there were statistical significantly differences （P〈0 .05） .Real‐time PCR results showed Gli1 of the cyclopamine treat‐ment group with the concentration of 5 μmol/L ,15 μmol/L ,25 μmol/L were cut down by 50 .4% ,61 .2% and 75 .7% on mRNA level that the there were also statistical significantly differences compared with the control group （P〈0 .05） .Conclusion The effect of cyclopamine can reduce the proliferation in HepG2 .2 .15 cells .The mechanism may be related to the mRNA expression of Gli 1 in HepG2 .2 .15 cells .

关 键 词：HEDGEHOG信号通路 肝癌 增殖 

分 类 号：R33[医药卫生—人体生理学] R392.12[医药卫生—基础医学]