二乙酰二去水卫矛醇对肺癌细胞NCI-H460增殖及凋亡的影响

摘要

目的 探讨二乙酰二去水卫矛醇(DADAG)对肺癌细胞NCI-H460增殖与凋亡的影响.方法 采用MTT法检测不同浓度DADAG(2.88、5.75、11.50、23.00、46.00 μg/mL)作用于NCI-H460细胞48 h后的增殖变化;集落形成实验检测DADAG对NCI-H460细胞抗癌活性的影响;AO/EB染色法于荧光显微镜下观察DADAG处理后细胞凋亡形态学的改变;流式细胞术检测DADAG对NCI-H460细胞凋亡的影响;RT-PCR检测DADAG对Bcl-2、Bax mRNA表达水平的影响.结果 与空白组比较,DADAG处理组细胞增殖能力降低,细胞形成集落的数量减少;AO/EB染色后细胞形态出现固缩、肿胀等典型凋亡形态学的改变;流式细胞仪检测结果显示DADAG处理组凋亡率较空白组高,差异有统计学意义(P<0.05);RT-PCR结果显示Bax表达上调,Bcl-2表达下调.结论 DADAG对肺癌细胞株NCI-H460的增殖有抑制作用,并能诱导其凋亡,其诱导凋亡可能与Bcl-2上调、Bax下调有关.%Objective To observe the effects of diacetyldianhydrogalactito (DADAG) on the proliferation and apopotosis in lung cancer cell NCI-H460.Methods MTT assay was performed to determine the half inhibitory concentration of DADAG(2.88,5.75,11.50,23.00 and 46.00 μg/mL)for the NCI-H460 cells and colony formation assay was used to detect the ability of cell proliferation; through AO/EB staining method, morphological changes of apoptotic cell under fluorescent microscope were observed;cell apoptosis was detected by flow cytometry to test the effects of DADAG on NCI-H460;RT-PCR was used to detect the effect of DADAG on Bcl-2 and Bax mRNA expres-sion levels within the cell. Results Compared with that in blank control group, cell proliferation was inhibited in the group treated with DADAG; the number of colony formation decreased and AO/EB staining results showed that cell apoptosis was characterized by typical morphological changes such as swelling, shrinking and fragmentation. Flow cytometry detection results showed that apoptosis rate was increased in the group treated with DADAG(all P<0.05).RT-PCR results indicated that the expression of pro-apoptosis gene Bax was up-regulated,while expression of anti-apoptosis gene Bcl-2 down-regulated. Conclusions DADAG exhibits the inhibitory activity in lung cancer cell NCI-H460 and can induce the apoptosis.The potential mechanism maybe associated with up-regulating the ex-pression of Bax but down-regulating the expression of Bcl-2.