雷帕霉素对S180肉瘤细胞及CD90、CD133、Notch1蛋白表达的影响  

Influence of rapamycin on the growth of sarcoma S180 cells and on the expression ofcluster of differentiation 90 (CD90),cluster of differentiation 133 (CD133) and neurogenic gene homologous protein 1 (Notch1)

作　　者：孔萍 王康嵘 张静 KONG Ping;WANG Kangrong;ZHANG Jing(Department of Pharmacy,Yichang Yiling Hospital,Yichang 443100,China;Department of Otolaryngology,Oral Head and Neck Surgery,Wuhan Puren Hospital,Wuhan 430081,China)

机构地区：[1]宜昌市夷陵医院药剂科,宜昌443100 [2]武汉市普仁医院耳鼻喉口腔头颈外科,武汉430081

出　　处：《西北药学杂志》2021年第4期595-600,共6页Northwest Pharmaceutical Journal

基　　金：湖北省卫生健康委员会联合基金项目(编号:WJ2019H153)。

摘　　要：目的探讨雷帕霉素对小鼠S180肉瘤细胞生长及抗原分化簇90(CD90)、抗原分化簇133(CD133)、神经源性基因同源蛋白(Notch1)蛋白表达的影响。方法选取处于对数生长期的S180肉瘤细胞,雷帕霉素低、中、高剂量组加入20、50、100 nmol·L-1雷帕霉素,对照组加入等量培养液,采用CCK-8法检测细胞增殖,流式细胞术法检测细胞周期与凋亡。将KM雄性小鼠随机分为对照组以及雷帕霉素低、中和高剂量组,每组10只。按照人体临床用量的0.5、1.0、2.0倍换算小鼠雷帕霉素用量后灌胃,对照组以等量生理盐水灌胃,每日1次,共3周,构建小鼠S180肉瘤皮下荷瘤模型。末次灌胃后24 h,测定小鼠体质量,记录瘤质量并计算抑瘤率。分别采用免疫荧光染色和免疫组织化学染色检测小鼠肿瘤组织CD90、CD133和Notch1的蛋白表达。结果雷帕霉素作用S180肉瘤细胞后,细胞增殖抑制率显著升高(P<0.05),且与作用时间呈正相关;同一作用时间,细胞增殖抑制率随雷帕霉素浓度的增加而升高(P<0.05);不同浓度雷帕霉素作用S180肉瘤细胞48 h后,细胞阻滞于G0/G1期,细胞凋亡率显著升高(P<0.05);免疫荧光染色结果显示,与对照组比较,其他3组小鼠肿瘤组织CD90、CD133和Notch1表达水平显著降低,且随雷帕霉素浓度增加而降低(P<0.05);免疫组织化学染色结果显示,与对照组比较,其他3组小鼠肿瘤组织CD90、CD133和Notch1表达水平显著降低,且随雷帕霉素浓度增加而降低(P<0.05)。结论雷帕霉素能通过下调CD90、CD133和Notch1蛋白表达影响S180肉瘤细胞的增殖和凋亡,达到体内外抑瘤作用。Objective To investigate the effects of rapamycin on the growth of sarcoma S180 cells and on the expression of cluster of differentiation 90(CD90),cluster of differentiation 133(CD133)and neurogenic gene homologous protein 1(Notch1)protein in mice model.Methods Sarcoma S180 cells at logarithmic growth stage were selected and cultured in the medium contained different concentration levels rapamycin(20,50,100 nmol·L-1),and the cells in control group were cultured in the same amount of medium.Cell proliferation,cell cycle and apoptosis were detected by CCK-8 and flow cytometry method.KM male mice were randomly divided into control group,rapamycin low,middle and high dose groups,each with 10 mice.Mice were given rapamycin by gavage,and the doses were converted according to 0.5,1.0 and 2.0 times of human clinical dosage.The control group was given the same amount of normal saline.The treatment in all 3 groups was once per day for 3 weeks.Tumor-bearing models were established by subcutaneously implanting S180 tumor cells to mice.24 hours after the last gavage,the weight of the mice was measured,the tumor weight was recorded,and the tumor inhibition rate was calculated.Immunofluorescence staining and immunohistochemical staining were used to detect CD90,CD133 and Notch1 protein expression in tumor tissues.Results After treatment with rapamycin at different concentrations on S180 sarcoma cells,the inhibition rate of cell proliferation was significantly increased(P<0.05),which was positively correlated with the action time.Under the same treatment time,the inhibition rate of cell proliferation increased with the increase of rapamycin concentration(P<0.05).After S180 sarcoma cells were treated with rapamycin at different concentrations for 48 hours,the cells were mainly arrested at G0/G1 phase,and the apoptosis rate was significantly increased(P<0.05).The results of immunofluorescence staining showed that the expression levels of CD90,CD133 and Notch1 in tumor tissues of the other three groups were significantly lower tha

关 键 词：雷帕霉素 S180肉瘤细胞 抑瘤作用 抗原分化簇90(CD90) 抗原分化簇133(CD133) 神经源性基因同源蛋白(Notch1) 

分 类 号：R965[医药卫生—药理学]