mTOR介导的细胞自噬在依维莫司治疗甲状腺癌中的作用及机制  

Role and Mechanism of mTOR-Mediated Autophagy in the Treatment of Thyroid Cancer with Everolimus

作　　者：王宁[1] 刘金彪[1] WANG Ning;LIU Jin-biao(Thyroid Surgery,the First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471003,China)

机构地区：[1]河南科技大学第一附属医院甲状腺外科,洛阳471003

出　　处：《中国合理用药探索》2021年第4期102-106,共5页Chinese Journal of Rational Drug Use

摘　　要：目的:探讨哺乳动物雷帕霉素靶蛋白(mTOR)介导的细胞自噬在依维莫司治疗甲状腺癌中的作用及机制。方法:选用NIH3T3"HOOK3-RET"稳定细胞株构建甲状腺肿瘤BALB/C裸鼠皮下移植模型,随机分为对照组、模型组和治疗组,每组30只。治疗组裸鼠造模成功后依维莫司(1.5 mg/kg)连续灌胃给药30天。测定甲状腺肿瘤裸鼠瘤体体积和重量;HE染色观察甲状腺肿瘤病理改变;免疫组织化学法检测组织中m TOR蛋白表达;采用Western Blot法检测甲状腺癌组织中m TOR、LC-3II、p62、Atg5和NF-κB蛋白表达。用不同浓度的依维莫司(0、5和10 nmol/L)处理SW579细胞株培养48 h,检测细胞存活率和细胞凋亡率。结果:模型组和治疗组裸鼠在细胞移植后逐渐成瘤,治疗前瘤体体积和重量比较无统计学差异(P> 0.05)。随着移植时间延长,模型组裸鼠的瘤体体积和重量逐渐增大(P <0.05);给予依维莫司后,治疗组裸鼠的瘤体体积和重量逐渐降低(P <0.05)。与对照组相比,模型组和治疗组LC-3Ⅱ、Beclin-1和Atg5蛋白水平降低,模型组mTOR、p62和NF-κB蛋白水平增加,具有统计学差异(P <0.05);与模型组相比,治疗组LC-3II、Beclin-1和Atg5蛋白水平升高,mTOR、p62和NF-κB蛋白水平降低(P <0.05)。低剂量组和高剂量组的SW579细胞存活率均低于对照组,而SW579细胞凋亡率均高于对照组;且随着依维莫司剂量的升高,SW579细胞存活率降低,SW579细胞凋亡率升高(P <0.05)。结论:m TOR抑制剂依维莫司可通过抑制m TOR介导的细胞自噬信号通路,有效治疗和抑制甲状腺肿瘤的生长与增殖。Objective: To investigate the role and mechanism of mammalian target of rapamycin( m TOR mediated autophagy in the treatment of thyroid cancer with everolimus. Methods: The BALB/C nude mice model of thyroid tumor was established by NIH3 T3 ‘HOOK3-RET’cell line and divided randomly into control group,model group and treatment group,with 30 mice in each group. After successful establishment of mice model,nude mice in treatment group were given everolimus( 1. 5 mg/kg) for 30 days through gavage. The volume and weight of thyroid tumor were determined. The pathological changes of thyroid tumors were observed through HE staining.Immunohistochemistry was used to detect the expression of mTOR protein in tissues. Western Blot was used to detect the expression of mTOR,LC-3 Ⅱ,p62,Atg5 and NF-κB in thyroid cancer tissues. SW579 cells were treated with different concentrations of everolimus( 0,5 and 10 nmol/L) for 48 h,and the cell viability and apoptosis rate were detected. Results: The nude mice in the model group and the treatment group gradually developed tumors after cell transplantation. There was no significant difference in tumor volume and weight before treatment( P > 0. 05).The tumor volume and weight of nude mice gradually increased with the time of transplantation( P < 0. 05). After the treatment with everolimus in nude mice,the volume and weight of tumor in nude mice in treatment group gradually decreased( P < 0. 05). Compared with the control group,the levels of LC-3 II,Beclin-1 and Atg5 in the model group and the treatment group were significantly decreased,while the levels of mTOR,p62 and NF-κB were significantly increased( P < 0. 05). Compared with the model group,the levels of LC-3 II,Beclin-1 and Atg5 in the treatment group were significantly increased,while the levels of m TOR,p62 and NF-κB were significantly decreased( P < 0. 05). The survival rate of SW579 cells in low-dose group and high-dose group was lower than that of the control group,while the apoptosis rate of SW579 cells was higher than

关 键 词：依维莫司 甲状腺癌 MTOR 自噬 凋亡 

分 类 号：R977[医药卫生—药品]