基于Fas/FasL信号通路探究雷帕霉素对食管癌裸鼠瘤体抑制及放疗增敏的作用机制  

Effect of rapamycin on inhibiting esophageal cancer tumor in mice and sensitizing radiotherapy based on Fas/FasL signaling pathway

作　　者：金元元 孟德胜 刘雅恬[1] JIN Yuanyuan;MENG Desheng;LIU Yatian(Jiangsu Cancer Hospital,Nanjing 210009,China)

机构地区：[1]江苏省肿瘤医院,江苏南京210009

出　　处：《现代药物与临床》2025年第1期15-22,共8页Drugs & Clinic

基　　金：江苏省卫生计生委科研课题项目(H2017036)。

摘　　要：目的探究雷帕霉素通过Fas/FasL信号通路治疗食管癌及放疗增敏的具体机制。方法采用CCK-8和流式细胞分析评价ECA109细胞的凋亡能力。采用ECA109 sc构建食管癌小鼠瘤体模型。将C57BL/6J小鼠分为对照组、放射组(4 Gy^(137)Csγ射线)、雷帕霉素(雷帕霉素1.5 mg/kg)组、放射+雷帕霉素(4 Gy^(137)Csγ射线+雷帕霉素1.5 mg/kg)组。收集并记录各组小鼠的肿瘤体积及质量、肿瘤抑瘤率及增殖系数,放疗及雷帕霉素对其的增敏影响。采用酶联免疫吸附试剂盒评估肿瘤组织中半胱氨酸蛋白酶3(Caspase-3)、B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)水平。苏木素–伊红(HE)染色观察小鼠食管癌的病理影响。Western blotting评估Fas/FasL信号通路的激活情况。结果与放射组相比,放射+雷帕霉素组小鼠肿瘤体积、质量显著减小(P<0.001)。与放射组相比,放射+雷帕霉素组的抑瘤率及生长延迟时间(TGD)显著升高,肿瘤3倍倍增时间(TGT3)显著延长(P<0.05)。放射+雷帕霉素组出现大量肿瘤细胞崩解,间质水肿更明显。与放射组比较,放射+雷帕霉素组Bcl-2表达水平显著降低,Bax、Caspase-3表达水平显著升高(P<0.001)。与放射组比较,放射+雷帕霉素组ECA109细胞活力显著降低(P<0.001)。Fas及FasL的表达在细胞中被抑制后,ECA109细胞活力显著增加,ECA109细胞凋亡率显著降低(P<0.001)。与放射组相比,放射+雷帕霉素组中Fas和FasL蛋白相对表达量显著升高(P<0.001)。结论雷帕霉素通过Fas/FasL信号通路抑制食管癌肿瘤的进展,该过程能有效地促进食管癌小鼠的放疗增敏。Objective To explore the specific mechanisms of rapamycin treating esophageal cancer,and sensitizing radiotherapy through the Fas/FasL signaling pathway.Methods The apoptosis ability of ECA109 cells was evaluated using CCK-8 and flow cytometry.The ECA109 mouse model of esophageal cancer was established using ECA109.C57BL/6J mice were divided into control group,radiation group(4 Gy^(137)Csγrays),rapamycin group(rapamycin 1.5 mg/kg),and radiation+rapamycin(4 Gy^(137)Csγrays+rapamycin 1.5 mg/kg).The tumor volume and mass,tumor inhibition rate and growth coefficient,the sensitization effect of radiotherapy and rapamycin were recorded.ELISA kits were used to assess levels of Caspase-3,Bcl-2,and Bax in the tumor.Histological changes in the esophageal cancer of mice were observed using HE staining.Western blotting analysis was performed to evaluate the activation of the Fas/FasL signaling pathway.Results Compared with the radiation group,the tumor volume and mass in the radiation+rapamycin group were significantly reduced(P<0.001).Compared with the radiation group,the tumor inhibition rate and TGD in the radiotherapy+rapamycin group were significantly increased,and the TGT3 was significantly prolonged(P<0.05).In the radiotherapy+rapamycin group,a large number of tumor cells disintegrated,and interstitial edema was more obvious.Compared with the radiation group,the expression levels of Bcl-2 in the radiation+rapamycin group were significantly decreased,while the expression levels of Bax and Caspase-3 were significantly increased(P<0.001).Compared with the radiation group,ECA109 cell viability was significantly decreased in the radiation+rapamycin group(P<0.001).After the expression of Fas and FasL was inhibited,the viability of ECA109 cells was significantly increased,and the apoptosis rate of ECA109 cells was significantly decreased(P<0.001).Compared with the radiation group,the relative expressions of Fas and FasL proteins in the radiation+rapamycin group were significantly increased(P<0.001).Conclusion Rapamycin inh

关 键 词：雷帕霉素 食管癌 放疗增敏 Fas/FasL信号通路 半胱氨酸蛋白酶3 B淋巴细胞瘤-2 Bcl-2相关X蛋白 

分 类 号：R965[医药卫生—药理学]