钌络合物通过诱发高尔基体应激在Walker-256荷瘤大鼠中发挥抗肿瘤作用的机制  

The Mechanism of Ruthenium Complex Exerting Anti-tumor Effect in Walker-256 Tumor-bearing Rats by Inducing Golgi Stress

作　　者：李量 吴文安 刘棣[2] 常浩 刘慧娟 刘佳[4] LI Liang;WU Wen-an;LIU Di;CHENG Hao;LIU Hui-juan;LIU Jia(Department of Radiotherapy,Shaanxi Cancer Hospital Affiliated to Xi'an Jiaotong University Health Science Center,Xi'an,Shaanxi,710061,China;Department of Oncology,The Second Affiliated Hospital Of Xi'an Jiaotong University,Xi'an,Shaanxi,710004,China;Department of Radiotherapy,Tangdu Hospital Affiliated to Air Force University,Xi'an Shaanxi,710038,China;Department of Thoracic Surgeiy,Shaanxi Cancer Hospital Affiliated to Xi'an Jiaotong University Health Science Center,Xi'an,Shaanxi,710061,China)

机构地区：[1]西安交通大学医学院附属陕西省肿瘤医院放疗科,陕西西安710061 [2]西安交通大学第二附属医院肿瘤科,陕西西安710004 [3]空军军医大学附属唐都医院放疗科,陕西西安710038 [4]西安交通大学医学院附属陕西省肿瘤医院胸外科,陕西西安710061

出　　处：《现代生物医学进展》2021年第23期4417-4421,4431,共6页Progress in Modern Biomedicine

基　　金：陕西省科技厅基金项目(2017JM8158)。

摘　　要：目的:探讨钌络合物通过诱发高尔基体应激在Walker-256荷瘤大鼠中发挥抗肿瘤作用的机制。方法:以30只雄性Wistar大鼠为研究对象,通过Walker-256细胞右骨盆肢体皮下注射建立荷瘤大鼠模型,然后根据实验目的将大鼠分为3组,对照组(正常大鼠,PBS干预),肿瘤模型组(荷瘤模型大鼠,PBS干预)和钌络合物组[荷瘤模型大鼠,管饲法给予5 mg/kg钌络合物溶液(由含2%Tween的PBS溶解)],各10只。通过测厚仪和电子秤分别计算大鼠肿瘤体积及重量;酶联免疫吸附试验试剂盒检测大鼠刚脏组织匀浆中氧化应激水平;蛋白印迹和荧光探针DCFH-DA试剂盒分析LC3 II/I表达和ROS活性;蛋白印迹分析高尔基应激相关蛋白GOLPH3、GRASP65的表达;实时定量PCR分析Bax、Bcl-2和Caspase-3的m RNA表达。结果:钌络合物组较肿瘤模型组肿瘤重量降低(P<0.05),肿瘤模型组较对照组体重增加降低(P<0.05),钌络合物组较肿瘤模型组体重增加(P<0.05)。肿瘤模型组较对照组SOD活性和LPO升高(P<0.05),CAT、GST和GSH活性降低(P<0.05),钌络合物组较肿瘤模型组LPO降低(P<0.05),CAT、GST和GSH活性升高(P<0.05)。肿瘤模型组较对照组LC3 II/I蛋白表达和ROS活性升高(P<0.05),钌络合物组较肿瘤模型组LC3 II/I蛋白表达和ROS活性降低(P<0.05)。肿瘤模型组较对照组GOLPH3、GRASP65的蛋白表达升高(P<0.05),钌络合物组较肿瘤模型组GOLPH3、GRASP65的蛋白表达降低(P<0.05)。肿瘤模型组较对照组Bax和Caspase-3的m RNA表达升高(P<0.05),Bcl-2 m RNA表达降低(P<0.05),钌络合物组较肿瘤模型组Bax和Caspase-3的m RNA表达降低,Bcl-2 m RNA表达升高(P<0.05)。结论:钌络合物通过调节高尔基应激反应,削弱氧化磷酸化从而促进Walker-256细胞死亡发挥抗肿瘤活性。Objective:To investigate the mechanism of ruthenium complexes exerting anti-tumor effects in Walker-256 tumor-bearing rats by inducing Golgi stress.Methods:Thirty male Wistar rats were used as research objects.Walker-256 cells were injected subcutaneously into the right pelvic limb to establish a tumor-bearing rat model,then divided into 3 groups according to the experimental purpose,the control group(normal rats,PBS intervention),tumor model group(tumor-bearing model rats,PBS intervention)and ruthenium complex group[tumor-bearing model rats,given 5 mg/kg ruthenium complex solution by gavage(dissolved in PBS containing 2%Tween)],10 each.The volume and weight of the rat tumor were calculated by the thickness gauge and the electronic scale;ELISA kit was used to detect the level of oxidative stress in rat visceral tissue homogenate;Western blot and fluorescent probe DCFH-DA kit were used to analyze LC3 II/I expression and ROS activity;Western blotting was used to analyze the expression of Golgi stress-related proteins GOLPH3 and GRASP65;RT-PCR was used to analyze the m RNA expression of Bax,Bcl-2 and Caspase-3 in real time.Results:The weight of the tumor in the ruthenium complex group was lower than that in the tumor model group(P<0.05),the weight of the tumor model group was lower than that in the control group(P<0.05),and the weight of the ruthenium complex group was higher than the tumor model group(P<0.05).Compared with the control group,the tumor model group had higher SOD activity and LPO activity(P<0.05),CAT,GST and GSH activity decreased(P<0.05),the ruthenium complex group had lower LPO than the tumor model group(P<0.05),CAT,The activities of GST and GSH increased(P<0.05).The tumor model group had higher LC3 II/I protein expression and ROS activity than the control group(P<0.05),and the ruthenium complex group had lower LC3 II/I protein expression and ROS activity than the tumor model group(P<0.05).The protein expression of GOLPH3 and GRASP65 in the tumor model group was higher than that in the control group

关 键 词：钌络合物 高尔基体 应激反应 抗肿瘤 大鼠 

分 类 号：R-33[医药卫生] R73-3