线粒体凋亡相关因子在大鼠自发性乳腺肿瘤中的表达  

Expression of mitochondrial apoptotic pathway-related factors in Sprague-Dawley rats with spontaneous mammary neoplasms

作　　者：张睿[1] 崔雨婷[1] 陈正礼[1] 罗启慧[1] 祝春梅[2] 孙凤娇 陈梦鹿 

机构地区：[1]四川农业大学动物医学院实验动物疾病模型研究室,四川雅安625014 [2]成都大学新药评价研究所,成都610051 [3]四川普莱美生物科技集团有限公司,四川雅安625014

出　　处：《中国实验动物学报》2015年第4期375-381,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家科技支撑计划课题(2014BAI03B01);国家重大科学仪器设备开发专项(2013YQ49085906);四川省青年科技创新研究团队项目(2013TD0015)

摘　　要：目的探讨SD大鼠自发性乳腺肿瘤中线粒体凋亡通路相关因子的表达情况及作用机制。方法将130只(♀∶♂=1∶1)Sprague-Dawley(SD)大鼠于SPF屏障环境中饲养,收集大鼠自发性乳腺肿瘤组织,通过病理学检查对肿瘤进行定性并分类,分为3组:正常乳腺组织(I组),良性乳腺肿瘤组织(II组)和恶性乳腺肿瘤组织(III组)。分别使用RT-PCR法和免疫组织化学法检测各样品中AIF、Cyt C、APAF-1、caspase-3、XIAP的mRNA和蛋白的表达。结果 130只大鼠中有14只检出自发性乳腺肿瘤,肿瘤发生率为10.77%;其中7只发生乳腺纤维腺瘤,7只发生乳腺癌,良、恶性肿瘤发生率均为5.38%。免疫组化结果如下,与I组相比,II组中AIF、APAF-1、caspase-3蛋白表达显著降低(P<0.01),Cyt C、XIAP蛋白表达显著增强(P<0.01),Ⅲ组中AIF、Cyt C、APAF-1、caspase-3蛋白表达均显著减弱(P<0.01),仅XIAP蛋白表达显著增强(P<0.01);与II组相比,III组中AIF、Cyt C、caspase-3蛋白表达显著减弱(P<0.01),APAF-1、XIAP蛋白表达显著增强(P<0.01)。RT-PCR结果中,除APAF-1的mRNA与蛋白表达显著相关(P<0.05),其余因子与免疫组化的结果极显著相关(P<0.01)。结论乳腺肿瘤是SD大鼠常见的肿瘤,线粒体凋亡通路相关因子AIF、Cyt C、APAF-1、caspase-3、XIAP在乳腺肿瘤中的异常表达与肿瘤的发生发展密切相关。Objective To investigate the expression of AIF,CYT C,PAF-1,caspase-3,and XIAP in SpragueDawley rats with spontaneous mammary neoplasms. Methods One-hundred and thirty 3-4-week old SPF Spargue-Dawley rats（ ♀∶ ♂ = 1∶ 1） were fed in a specific pathogen free（ SPF） breeding barrier for 60 weeks. The occurrence of spontaneous breast tumors was recorded and histopathology was performed to identify the types of tumors. The rats were divided into3 groups： rats with normal breast tissue（ group I）,with benign tumors（ group II） and with malignant tumors（ group III）.The expression of AIF,CYT C,APAF-1,caspase-3 and XIAP proteins and mRNAs were detected by immunhistochemistry（ IHC） and RT-PCR assay. Results Among these 130 SD rats,14 rats were observed having spontaneous mammary neoplasms with the incidence rate of 10. 77%（ 14 /130）. In these neoplasm cases,7 cases were mammary fibroadenomas,7cases of breast carcinoma,both with an incidence rate of 5. 38%. Immunohistochemistry showed that,compared with the group I,the positive expressions of AIF,APAF-1,caspase-3 were decreased significantly（ P〈0. 01）,and the CYT C and XIAP expressions were significantly increased in the group II. The positive expression of all genes except XIAP was decreased in the group III（ P〈0. 01）. Compared with the group II,APAF-1 and XIAP were significantly higher in the group III（ P〈0. 01）,and the positive expression of AIF,Cyt C,and caspases-3 were significantly decreased（ P〈0. 01）. In the results of RT-PCR assay,except APAF-1 which showed significant correlation with the results of immunohistochemistry（ P〈0. 05）,all the others showed an extremely significant correlation with immunohistochemical results（ P〈0. 01）.Conclusions Mammary tumors are most common spontaneous neoplasms in SD rats. Abnormal expression of mitochondrial apoptotic pathway-related factors AIF,CytC,APAF-1,caspase-3,and XIAP are correlated with the carcinogenesis and development of breast tumors.

关 键 词：自发乳腺肿瘤 SD大鼠 RT-PCR 免疫组化 肿瘤发生 

分 类 号：Q95-33[生物学—动物学]