摘要
目的探讨萝卜硫素对宫颈癌He La细胞凋亡及放射敏感性的影响。方法体外培养宫颈癌HeLa细胞,采用四氮唑比色(MTT)法确定萝卜硫素和放射线对He La细胞的干预剂量;采用克隆形成实验观察萝卜硫素对He La细胞的放射增敏作用;将HeLa细胞分为对照组、联合组、萝卜硫素组和放射组,检测细胞凋亡和细胞周期变化,同时检测He La细胞中细胞外信号调节激酶(ERK1/2)、磷酸化ERK1/2(p-ERK1/2)、Akt、磷酸化Akt(p-Akt)、半胱氨酸蛋白酶-3(Caspase-3)蛋白表达。结果随着萝卜硫素浓度和放射线剂量的增加,He La细胞抑制率增加,萝卜硫素对He La细胞的半数抑制浓度(IC50)为(76.28±9.27)μmol/L,故选80μmol/L作为干预浓度;放射线对HeLa细胞的IC50为(2.13±0.27)Gy,故选2 Gy作为半干预剂量;随着放射剂量的增加,放射组和联合组细胞存活率均降低,联合组各放射剂量的细胞存活率均低于放射组(P<0.05),萝卜硫素能提高HeLa细胞放射敏感性,SERD0和SERDq分别为1.517和2.024。各组He La细胞G0/G1期细胞比例变化不显著(P>0.05);与对照组比较,其余各组HeLa细胞凋亡率、G2/M期细胞比例、ERK1/2、p-ERK1/2、Akt、p-Akt、Caspase-3蛋白表达增加,S期细胞比例降低,联合组作用最强,放射组次之,萝卜硫素组最弱(P<0.05)。结论萝卜硫素可通过G2/M期阻滞诱导HeLa细胞凋亡,并通过激活ERK和Akt,增强He La细胞的放射敏感性。
Objective To investigate the effect of sulforaphane on apoptosis and radiosensitivity of He La cells of cervical cancer.Methods He La cells were cultured in vitro.The inhibitory dose of sulforaphane and radiation on He La cells was determined by methyl thiazolyl tetrazolium(MTT)method.The radiosensitization effect of sulforaphane on HeLa cells was observed by clonogenesis experiment.HeLa cells were divided into the control group,combined group,sulforaphane group and radiation group.Cell apoptosis and cell cycle changes were detected.The expression of extracellular signal regulated kinase 1/2(ERK1/2),p-ERK1/2,AKT,p-Akt and Caspase-3 protein in He La cells were detected.Results With the increase of sulforaphane concentration and radiation dose,the inhibition rate of He La cell increased.The half inhibitory concentration(IC50)of sulforaphane on He La cells was(76.28±9.27)μmol/L,so 80μmol/L was selected as the intervention concentration.The half inhibitory concentration(IC50)of He La cells by radiation was(2.13±0.27)Gy,so 2 Gy was chosen as the semi-intervention dose.With the increase of radiation dose,the cell survival rate of radiation group and combined group was significantly decreased,and the survival rate of HeLa cells in the combined group was lower than that in the radiation group(P<0.05).Sulforaphane could improve the radiosensitivity of He La cells,and SERD0 and SERDq were 1.517 and 2.024 respectively.The proportion of G0/G1 phase in HeLa cells did not significantlychanged in each group(P>0.05).Compared with the control group,the apoptosis rate,G2/M cell proportion,ERK1/2,p-ERK1/2,Akt,p-Akt and Caspase-3 protein expression in HeLa cell of other groups were significantly increased,the proportion of cells in S phase decreased,and the combined group had the strongest effect,followed by radiation group,and the effect of sulforaphane group was the weakest(P<0.05).Conclusion Sulforaphane can induce HeLa cells apoptosis through G2/M phase arrest,and enhance the radiosensitivity of HeLa cells through the activation of ERK and Akt.
作者
张亚芳
赵晓红
王德珠
张华凤
王小花
ZHANG Yafang;ZHAO Xiaohong;WANG Dezhu;ZHANG Huafeng;WANG Xiaohua(Department of Gynecology and Obstetrics,Hainan Womenand Children Medical Center,Haikou,Hainan,China 570206)
出处
《中国药业》
CAS
2020年第15期22-26,共5页
China Pharmaceuticals
