二去水卫矛醇对人肺癌细胞株体外抗癌活性及机制探讨  

In Vitro Anticancer Activity and Mechanisms of 1,2: 5,6-Dianhydrogalactitol on Human Lung Cancer Cell Lines

作　　者：苏桂玉 刘华钢[2] 黄慧学[1] 彭晓丽[1] 梁乔芳 徐红娟[1] 王小洁[2] 

机构地区：[1]广西中医药大学药学院,南宁530299 [2]广西医科大学药学院,南宁530021

出　　处：《中国实验方剂学杂志》2016年第10期122-127,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：广西科学研究与技术开发计划项目(桂科能12237007)

摘　　要：目的:研究二去水卫矛醇(DAG)对人肺癌细胞株的体外增殖及凋亡的影响。方法:Cell Counting Kit-8(CCK-8)法检测DAG对11株人肺癌细胞株Calu-1,NCI-H1650,NCI-H358,NCI-H1299,HCC827,PC-9,A549,NCI-H661,NCI-H292,95-D和NCI-H446的体外增殖抑制率,筛选出抑制率相对较高且生长状态良好的细胞;应用台盼蓝拒染法检测DAG作用后的细胞存活率,透射电镜观察细胞凋亡亚显微结构的改变,实时定量荧光PCR(Real-time PCR)检测DAG对细胞内B淋巴细胞瘤-2(Bcl-2),Bcl-2相关X蛋白(Bax)和半胱氨酸蛋白酶-3(Caspase-3)mRNA表达水平的影响。结果:DAG对11株肺癌细胞株均有明显的抑制作用,并呈现出良好的浓度-效应相关性,与空白组比较,随着浓度的增加,细胞的增殖抑制率上升(P<0.01);台盼蓝拒染实验显示随着药物浓度的增加,蓝染细胞数量增多,即死亡或细胞膜遭到破坏的细胞增多;药物作用48 h以后,透射电镜下可见凋亡细胞整体圆缩,微绒毛脱落,核固缩、边集,核内出现高电子密度的异染色质,浓缩成块状,边集于核膜,线粒体肿胀,细胞内出现空泡样变;Real-time PCR结果显示,促凋亡基因Bax表达上调,抗凋亡基因Bcl-2表达下调,Caspase-3被激活。结论:DAG能够抑制肺癌细胞的增殖,并诱导其凋亡,其作用机制可能与上调Bax,下调Bcl-2,激活Caspase-3有关。Objective： To investigate the effect of 1,2： 5,6-dianhydrogalactitol（ DAG） in vitro proliferation and apoptosis of human lung cancer cell lines. Method： Cell counting Kit-8（ CCK-8） method was used to measure the inhibition rate of DAG in vitro proliferation of 11 kinds of human lung cancer cells： Calu-1,NCI-H1650,NCI-H358,NCI-H1299,HCC827,PC-9,A549,NCI-H661,NCI-H292,95-D and NCI-H446,then cells with higher inhibition rates and good growth conditions were selected for subsequent research. Trypan blue exclusion method was used to detect the cell survival rate after treatment by DAG. Transmission electron microscope was used to observe the changes in submicroscopic structure after apoptosis. Real-time PCR was used to detect the effect of DAG on Bax,Bcl-2 and Caspase-3 mRNA expression levels within the cells. Result： DAG significantly inhibited the 11 kinds of human lung carcinoma cell lines with a good concentration-effect relationship.As compared with the blank group,the inhibition rate on cells proliferation was increased with the increase of dose（ P〈0. 01）. Trypan blue exclusion results showed that,with the increase of drug concentrations,the count of cells in blue（ dead cells or cells with damaged membrane） was increased. Transmission electron microscope results indicated that,after 48 h treatment,apoptotic cells were rounded overall,microvilli fell off,with karyopyknosis and margination, heterochromatin with high electron density in the nucleus was condensed into lumps and aggregated on nuclear membranes,with mitochondrial swelling and vacuolar degeneration within cells. Real-time PCR results indicated that the expression of pro-apoptosis gene Bax was up-regulated,expression of anti-apoptosis gene Bcl-2 was down-regulated,and Caspase-3 was activated. Conclusion： DAG can inhibit the human lung carcinoma cells growth and induce their apoptosis, and the potential mechanism may be associated with upregulating the expression of Bax,down-regulating the expression of Bcl-2 and activating

关 键 词：二去水卫矛醇 人肺癌细胞 抗癌活性 

分 类 号：R285.5[医药卫生—中药学]