摘要
目的探讨了雷帕霉素(RAPA)对荷RM-1前列腺癌小鼠肿瘤生长的影响以及抗肿瘤机制.方法不同浓度的RAPA处理RM-1细胞,分析自噬蛋白和凋亡蛋白变化.将40只荷RM-1前列腺癌小鼠随机分为模型组、5、10、20 mg/kg RAPA组.5、10、20 mg/kg RAPA组大鼠通过腹腔分别注射RAPA 5、10、20 mg/kg.模型组腹腔注射相同体积的磷酸盐缓冲液(PBS).连续治疗30 d,分别观察3组小鼠肿瘤生长和生存率的情况.采用蛋白质印迹法(Western blot)分析3组小鼠肿瘤组织中半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3和p62蛋白质水平,并采用原位缺口末端标记法(TUNEL)染色法分析肿瘤组织细胞凋亡情况.应用SPSS 17.0统计学软件进行分析,计量数据采用均值±标准差(Mean±SD)表示,组间计量资料比较采用方差分析.结果与对照组(1.89±0.17)比较,125、250、500 nmol/L(0.91±0.19、0.68±0.12、0.31±0.10)处理的细胞p62水平明显下降,差异有统计学意义(t=2.019、3.491、4.910,P<0.05).与对照组(0.21±0.09)比较,125、250、500 nmol/L(0.41±0.14、0.87±0.18、2.01±0.21)处理的细胞Caspase-3表达水平显著增加,差异有统计学意义(t=1.989、2.582、5.019,P<0.05).与模型组比较,5、10、20 mg/kg RAPA组小鼠肿瘤生长明显减缓,且生存率较模型组明显提高,并呈剂量依赖性.与模型组比较,5、10、20 mg/kg RAPA组小鼠肿瘤组织中Caspase-3水平明显增加,而p62蛋白水平明显下降.5、10、20 mg/kg RAPA组Casepase-3和p62蛋白与RAPA浓度呈剂量依赖性.与模型组(5.01±1.12)比较,3、6、9 mg/kg RAPA组(20.14±4.12、51.23±8.99、78.43±10.32)小鼠肿瘤细胞的凋亡水平显著增加,差异有统计学意义(t=3.109、5.192、7.102,P<0.05).结论RAPA能够诱导肿瘤细胞发生自噬,而过度自噬可导致肿瘤细胞凋亡,进而达到治疗肿瘤的目的.
Objective To investigate the effect of rapamycin(RAPA)on the tumor growth of bearing cancer mice of RM-1 and the underlying mechanism.Methods In cellular level,RM-1 cells were treated with different concentrations of RAPA.The expression of p62 and cysteinyl aspartate-specific protease(Caspase)-3 proteins was detected by Western blotting.In Animal level,30 bearing cancer mice of RM-1 were randomly divided into model group,5 mg/kg RAPA group,10 mg/kg RAPA group and 20 mg/kg RAPA group.Mice in 5 mg/kg RAPA,10 mg/kg RAPA and 20 mg/kg RAPA groups were given intraperitoneal injection of RAPA with the doses of 5,10 and 20 mg/kg respectively.Mice in mode group were given the same volume of phosphate buffer(PBS).Tumor growth and survival rate of four groups were observed after administration for 30 days.Caspase-3 mRNA and protein levels were analyzed by real-time fluorescent quantitative polymerase chain reaction(FQ-PCR)and Western blotting respectively.Cell apoptosis index in tumor tissue was analyzed by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining method.Results As compared with the control group(1.89±0.17),the levels of p62 in the cells treated with 125 nmol/L,250 nmol/L and 500 nmol/L(0.91±0.19,0.68±0.12,0.31±0.10)decreased significantly(t=2.019,3.491,4.910,P<0.05).As compared with the control group(0.21±0.09),the expression levels of Caspase-3 protein in cells treated with 125 nmol/L,250 nmol/L and 500 nmol/L were significantly increased(0.41±0.14,0.87±0.18,2.01±0.21)(t=1.989,2.582,5.019,P<0.05).As compared with the model group,RAPA of 5,10 and 20 mg/kg significantly slowed down the growth of tumors in mice,and the survival rate was significantly higher than that in the model group in a dose-dependent manner.As compared with the model group,the levels of Caspase-3 and p62 proteins in tumor tissues of mice in 5 mg/kg RAPA,10 mg/kg RAPA and 20 mg/kg RAPA groups increased significantly,and those of p62 protein decreased significantly.Casepase-3 and p62 proteins in 5 mg/kg RAPA,
作者
张士龙
陈国晓
朱晓博
单磊
张祥生
Zhang Shilong;Chen Guoxiao;Zhu Xiaobo;Shan Lei;Zhang Xiangsheng(Department of Urology,the People’s Hospital of Zhengzhou University,Henan Provincial People’s Hospital,Zhengzhoou 450003,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第9期1626-1629,共4页
Chinese Journal of Experimental Surgery
