7-二氟亚甲基-5,4'-二甲氧基染料木黄酮对宫颈癌细胞增殖和凋亡的影响及其机制  

Effects of 7-difluoromethy-5,4'-dimethoxygenistein on proliferation and apoptosis of human cervical cancer cells and its mechanism

作　　者：陈砚芬[1] 白军 徐家丽[1] 宋晓晖[1] 

机构地区：[1]武汉市妇女儿童医疗保健中心妇产科,武汉430000 [2]暨南大学第一临床医学院妇产科,广州510632

出　　处：《中南大学学报（医学版）》2016年第5期463-470,共8页Journal of Central South University ：Medical Science

摘　　要：目的：检测7-二氟亚甲基-5,4＇-二甲氧基染料木黄酮（7-difluoromethoxy-5,4＇-dimethoxygenistein,DFMG）对宫颈癌HeLa细胞增殖和凋亡的影响并探讨其可能机制。方法：体外培养宫颈癌HeLa细胞,MTT法检测HeLa细胞的增殖;AO/EB染色检测HeLa细胞凋亡形态学改变;FCM检测HeLa细胞凋亡率;DNA琼脂糖电泳检测HeLa细胞凋亡的DNA条带;RT-PCR和Western印迹检测DFMG对HeLa细胞凋亡相关蛋白表达的影响。结果：DFMG在体外呈浓度（0.25~64μg/mL）和时间（24~72 h）依赖性抑制HeLa细胞增殖,其作用48 h的IC_（50）值为4.62μg/mL,HeLa细胞出现典型的凋亡形态学改变,典型的凋亡DNA条带,凋亡率呈浓度依赖性增高,伴随c-myc mRNA和蛋白表达增高,其下游蛋白bax,cyto-c,caspase-9表达增高,而bcl-2蛋白表达降低。用siRNA干扰沉默c-myc基因,能部分抵消DFMG对HeLa细胞增殖抑制和凋亡诱导效应,而用c-myc cDNA转染HeLa细胞,则能协同DFMG对细胞增殖的抑制作用及对凋亡的诱导作用。结论：DFMG在体外具有抑制HeLa细胞增殖和诱导凋亡作用,其机制可能与钝化c-myc基因,启动线粒体凋亡途径有关。Objective：To investigate the effects of 7-difluoromethy-5,4- dimethoxygenistein（DFMG） on inhibiting proliferation and inducing apoptosis of human cervical cancer HeLa cells and its possible molecular mechanism in vitro.Methods：HeLa cells were cultured in vitro.The effect of DFMG on inhibiting proliferation was determined using MTT assay.The effects of DFMG on inducing apoptosis were assessed using flow cytometry with AV-PI staining,AO/EB staining,and agarose gel electrophoresis.Multiple molecular techniques,such as RT-PCR,Western blot,siRNA transfection,and cDNA transfection,were used to explore its possible molecular mechanism.Results：DFMG presented with dramatically inhibiting proliferation effect of HeLa cells in a timeand dose-dependent manner ranging from 0.25 to 64 μg/mL and from 24 to 72 h in vitro,and its IC_（50） was 4.62 μg/mL for 48 h.The cells treated with DFMG for 48 h showed typical morphological change of apoptosis,typical DNA ladder of agarose gel electrophoresis,and the sub-G_1 population increased in a dose-dependent manner.Simultaneously the expressions of c-myc mRNA,c-myc protein and its downstream genes,such as bax,cyto-c and caspase-9,were up-regulated,while bcl-2 protein was down-regulated.Down-regulation of c-myc by siRNA attenuated DFMG-induced cell proliferation inhibition and inducing apoptosis.Up-regulation expression of c-myc by cDNA transfection could enhance the effects of DFMG-induced cell proliferation inhibition and inducing apoptosis.Conclusion：DFMG could inhibit the proliferation and induce the apoptosis of human cervical cancer HeLa cells in vitro,and its mechanism may be closely related to regulate c-myc and its downstream gene expression.

关 键 词：宫颈癌 7-二氟亚甲基-5 4'-二甲氧基染料木黄酮 增殖 凋亡 C-MYC 

分 类 号：R737.33[医药卫生—肿瘤]