ROS1蛋白酪氨酸激酶抑制剂对卵巢癌移植瘤裸鼠的治疗作用及机制  

Therapeutic effect and mechanism of ROS1 protein tyrosine kinase inhibitor on ovarian cancer transplanted tumor in nude mice

作　　者：代雪梅 谢珊莉 贾凌彦 向德生 DAI Xue-mei;XIE Shan-li;JIA Ling-yan(Department of Gynecology,the First People's Hospital of Guangyuan,Guangyuan Sichuan 628000,China)

机构地区：[1]广元市第一人民医院妇科,四川广元628000

出　　处：《临床和实验医学杂志》2021年第24期2581-2585,共5页Journal of Clinical and Experimental Medicine

基　　金：四川省卫生厅科研课题(编号:20170713)。

摘　　要：目的探究ROS1蛋白酪氨酸激酶抑制剂对卵巢癌移植瘤裸鼠的治疗作用及机制。方法12只无特定病原体级5~7周龄的雌性无胸腺裸鼠采用腹腔注射人卵巢癌细胞建立移植瘤模型,成模后根据随机数字表法分为两组:模型组和实验组,分别用50 mg/kg PF-2341066和等量0.9%氯化钠溶液进行灌胃,治疗持续35 d后,分析肿瘤负荷以确定肿瘤总重量和转移数目。人卵巢癌Hey A8、SKOV3和Ca OV3细胞系分别采用浓度为0、0.025、0.05、0.10、1.00μmol/L的PF-2341066干预。蛋白质印迹法检测干预24 h后细胞中c-Met、Akt和Erk蛋白及其磷酸化蛋白相对表达量。以最佳作用剂量PF-2341066干预细胞,干预48 h后,通过用荧光分光光度计测量荧光强度量化黏附细胞的数量。分别干预24、48和72 h后,使用Cy Quant细胞增殖测定试剂盒测量卵巢癌细胞的增殖。结果与模型组小鼠相比,治疗组小鼠转移灶的数量和肿瘤总重量明显降低,存活时间显著延长,差异均有统计学意义(P<0.05)。不同浓度PF-2341066干预后,c-Met、Akt和Erk蛋白及其磷酸化蛋白相对表达量差异均有统计学意义(P<0.05),其大体趋势表现为随着PF-2341066浓度增加,c-Met蛋白及其磷酸化蛋白相对表达量显著降低,Akt和Erk蛋白及其磷酸化蛋白相对表达量显著增加,差异均有统计学意义(P<0.05)。择0.05μmol/L为最佳剂量进行干预,干预24 h后,SKOV3和Hey A8细胞系中Ⅰ型胶原蛋白、纤维连接蛋白和玻连蛋白表达均显著降低,差异均有统计学意义(P<0.05)。干预72h,SKOV3和Hey A8细胞细胞增殖能力显著降低,差异均有统计学意义(P<0.05)。结论PF-2341066在卵巢癌转移的体内模型中通过抑制c-Met、Akt和Erk蛋白及其磷酸化蛋白表达,从而抑制肿瘤黏附和增殖并延长移植瘤小鼠生存时间,因此可能是治疗卵巢癌患者的良好候选药物。Objective To explore the therapeutic effect and mechanism of ROS1 protein tyrosine kinase inhibitor on ovarian cancer transplanted nude mice.Methods Twelve specific pathogen free female athymic nude mice aged 5 to 7 weeks were intraperitoneally injected with human ovarian cancer cells to establish transplanted tumor models.After modeling,they were divided into 2 groups according to the random number table method: model group and experimental group,and the mice were intragastrically administered with 50 mg/kg PF-2341066 and the same amount of normal saline.After the treatment lasted 35 days,the tumor burden was analyzed to determine the total tumor weight and the number of metastases.Human ovarian cancer HeyA8,SKOV3 and CaOV3 cell lines were treated with PF-2341066 at concentrations of 0,0.025,0.05,0.10,and 1.00 μmol/L,respectively.The relative expressions of c-Met,Akt and Erk proteins and their phosphorylated proteins in the cells after 24 h of intervention were detected by Western blotting.Then intervene the cells with the optimal dose of PF-2341066,after 48 hours of intervention,the number of adherent cells was quantified by measuring the fluorescence intensity with a fluorescence spectrophotometer.After 24,48 and 72 hours of intervention,the proliferation of ovarian cancer cells were measured by CyQuant cell proliferation assay kit.Results Compared with mice in the model group,the number of metastases and the total tumor weight of the mice in the treatment group were significantly reduced,and the survival time was significantly prolonged,and the differences were statistically significant(P < 0.05).After the intervention of different concentrations of PF-2341066,the relative expressions of c-Met,Akt and Erk proteins and their phosphorylated proteins were significantly different(P < 0.05).The general trend showed that as the concentration of PF-2341066 increased,the relative expression of c-Met protein and its phosphorylated protein decreased significantly,and the relative expression of Akt and Erk protein and

关 键 词：裸鼠 卵巢癌 ROS1 基因 蛋白酪氨酸激酶抑制剂 基质金属蛋白酶 

分 类 号：R737.31[医药卫生—肿瘤]