摘要
目的观察双氢青蒿素(DHA)对宫颈癌小鼠放疗的增敏作用,并探讨其作用机制。方法选取小鼠宫颈癌细胞株U14常规培养、消化传代,取对数生长期U14细胞备用。取50只昆明小鼠,建立宫颈癌小鼠模型,将建模成功的小鼠随机分为模型组、放疗组、低剂量组(Low-dose,L)、中剂量组(Middle-dose,M)、高剂量组(High-dose,H)、DHA组。除模型组和DHA组均给予放疗,其中L组、M组、H组分别取5、10、20 mg/kg小鼠体质量的DHA+0.01 ml/g小鼠体质量的生理盐水混合均匀,DHA组给予5 mg/kg DHA+0.01 ml/g小鼠体质量的生理盐水混合均匀,均给予腹腔注射;放疗组和模型组给予0.01 ml/g小鼠体质量的生理盐水腹腔注射,均于照射前30 min干预。对比各组1周、2周、3周后抑瘤率变化;末次放疗次日将小鼠处死,以苏木精-伊红法(HE)染色观察各组肿瘤组织病理表现;以末端脱氧核苷酸转移酶介导的生物素脱氧尿嘧啶核苷酸缺口末端标记法(TUNEL)检测肿瘤组织细胞凋亡率;以荧光定量聚合酶链反应(RT-PCR)检测肿瘤组织磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)信使核糖核酸(mRNA)表达;以蛋白质免疫印迹(WB)检测PI3K、AKT蛋白及磷酸化-AKT(p-AKT)表达,计算p-AKT/AKT。结果共有46只小鼠建模成功;抑瘤率时间、组间、交互作用上差异均有统计学意义,各组抑瘤率均随时间的延长显著升高(P<0.05),L组、M组和H组抑瘤率均高于放疗组和DHA组(P<0.05),M组和H组抑瘤率均高于L组(P<0.05),H组抑瘤率高于M组(P<0.05),放疗组和DHA组差异均无统计学意义;模型组可见肿瘤细胞排列紊乱、大小不均、染色深浅不同,偶见细胞轻度水肿,间质存在少量的炎性细胞浸润;放疗组和DHA组均可见部分肿瘤细胞坏死且细胞呈水肿状态,核固缩,深浅不同,间质有明显水肿;L组可见肿瘤细胞区域性坏死,接近坏死处的肿瘤细胞体积变小,核固缩,有明显的间质水肿,可见炎性细胞浸润
Objective To observe the radiosensitizing effect of dihydroartemisinin(DHA)on mice with cervical cancer and explore its mechanism.Methods The mouse cervical cancer cell line U14 was selected for routine culture,digestion and passage,and the logarithmic growth phase U14 cells were taken for reserve.50 Kunming mice were selected to establish cervical cancer model.The mice were randomly divided into model group,radiotherapy group,low-dose(L)group,middle-dose(M)group,high-dose(H)group and DHA group.All mice except model group and DHA group were given radiotherapy.In group L,M and H,DHA of 5,10 and 20 mg/kg+0.01 ml/g normal saline mice were mixed evenly,and DHA of 5 mg/kg+0.01 ml/g normal saline mice in DHA group were mixed evenly before intraperitoneal injection.Radiotherapy group and model group were given intraperitoneal injection of 0.01 ml/g normal saline,which were intervened for 30 min before irradiation.We compared the changes of tumor inhibition rates after 1 week,2 weeks and 3 weeks in each group.The mice were executed the next day after the last chemotherapy,and the histopathological manifestations of the tumors were observed by hematoxylin-eosin(HE)staining.TdT-mediated dUTP Nick-End Labeling(TUNEL)was used to detect the apoptotic rates of tumor cells.The message ribonucleic acid(mRNA)expressions of Phosphatidylinositol 3-kinase(PI3K)and Protein kinase B AKT(AKT)were detected by reverse transcription-polymerase chain reaction(RT-PCR).Western Blot(WB)was used to detect the expression of PI3K and AKT proteins and phosphorylation-AKT(p-AKT),and we calculated p-AKT/AKT.Results A total of 46 mice were successfully modeled.There were significant differences in the time of tumor inhibition,the interaction among groups(P<0.05).The tumor inhibition rates of each group increased significantly with time(P<0.05).The tumor inhibition rates of L,M and H groups were higher than those of radiotherapy group and DHA group(P<0.05).The tumor inhibition rates of M and H groups were higher than that of L group(P<0.05).The tumor
作者
王佳茜
肖黎明
葛妍圻
WANG Jiaqian;XIAO Liming;Ge Yanqi(Department of Pharmaceutical Affairs,Beijing Maternity Hospital,Capital Medical University,Beijing 100026,China)
出处
《中国生育健康杂志》
2020年第4期328-334,I0001,共8页
Chinese Journal of Reproductive Health
关键词
双氢青蒿素
宫颈癌
放疗
增敏作用
机制
dihydroartemisinin
cervical cancer
radiotherapy
sensitization
mechanisms
