摘要
目的探讨E6相关蛋白（E6-AP）调节膜联蛋白A2的表达,对三阴性乳腺癌（TNBC）裸鼠移植瘤的影响。方法于2014年4~10月,选择40只雌性BALB/C-nu裸鼠为研究对象。按照随机数字表法,将其分为实验组（n=20）与对照组（n=20）。两组TNBC裸鼠移植瘤模型构建：设计针对E6-AP-RNA干扰靶点序列,将小干扰RNA（siRNA）片段转染至TNBC细胞株MDA-MB-231,再将此细胞株皮下注射于实验组裸鼠;而对照组裸鼠则皮下注射等量未转染E6-AP siRNA的TNBC细胞株MDA-MB-231。观察两组裸鼠移植瘤成瘤情况。采用Western blotting法检测两组TNBC裸鼠移植瘤E6-AP及膜联蛋白A2的相对表达量,采用免疫组化法检测两组TNBC裸鼠移植瘤膜联蛋白A2阳性表达率,并对这些指标进行统计学分析。本研究实验过程中对动物处置符合动物伦理学标准。两组裸鼠的周龄及体重等资料比较,差异均无统计学意义（P〉0.05）。结果 1成功构建两组TNBC裸鼠移植瘤模型,成瘤率均为100%。与对照组相比,实验组裸鼠移植瘤生长较慢,成瘤体积较小。实验组裸鼠移植瘤重量为（0.562±0.032）g,显著低于对照组的（1.202±0.012）g,二者比较,差异有统计学意义（t=96.792,P〈0.001）。2Western blotting检测结果显示,实验组E6-AP相对表达量为11 168±441,显著低于对照组的24 284±636,二者比较,差异有统计学意义（t=75.737,P〈0.001）。实验组膜联蛋白A2相对表达量为8 631±651,亦显著低于对照组的17 976±510,二者比较,差异有统计学意义（t=50.515,P〈0.001）。3免疫组化检测结果显示,实验组膜联蛋白A2阳性表达率为33%,明显低于对照组的58%,二者比较,差异有统计学意义（χ2=5.796,P=0.016）。实验组TNBC裸鼠移植瘤膜联蛋白A2的积分光密度（IOD）值为17 976±239,显著低于对照组的26 565±329,二者比较,差异亦有统计学意义（t=94.439,P〈0.001）。结论在人体内,膜联蛋白A2可能通过E6-AP介导发
Objective To study the influence of annexin A2 expression regulated by E6-associated protein（E6-AP）on trip-negative breast cancer（TNBC）in nude mouse with transplantation tumor.Methods From April to November 2014,40 female BALB/C-nu nude mice were selected as research subjects.They were divided into experiment group（n=20）and control group（n=20）according to random number table method.The construction of TNBC nude mice with transplantation tumor models in two groups： E6-AP-RNA interference target sequences were designed,and then small interfering RNA（siRNA）fragments were transfected into TNBC cell lines MDA-MB-231,and these TNBC cell lines were injected subcutaneously into nude mice in experimental group.While TNBC cell lines MDA-MB-231 which were not transfected with E6-AP siRNA were injected subcutaneously into control group.The transplanted tumor formations in two groups were observed.Relative expressions of E6-AP and annexin A2 in two groups were assayed through Western blotting method.And positive expression rate of annexin A2 in transplanted tumor of two groups were assayed by immunohistochemical method.And those indexes above were analyzed by statistical methods.The disposal of animals in this research was in accordance with the animal ethical standards.There were no significant differences between two groups in week of age and body weight（P〉0.05）.Results 1The TNBC nude mice with transplanted tumor models were constructed successfully in both two groups,and the tumor formation rates both were 100%.The growth of transplantation tumor in experimental group was relatively slower and the tumor volume was smaller than those of control group.The weight of transplantation tumor in experimental group was（0.562±0.032）g,which was significantly heavier than that of（1.202±0.012）g in control group,and the difference was statistical significant（t=96.792,P〈0.001）.2According to the results of Western blotting assay,the relative expression of E6-AP in experimental group was 11 168±
出处
《中华妇幼临床医学杂志（电子版）》
CAS
2016年第3期274-279,共6页
Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition)
基金
国家自然科学基金项目(81172496)
川北医学院课题项目基金项目(CBY12-A-QN07)~~
关键词
膜联蛋白A2
E6相关蛋白
裸鼠移植瘤
印迹法
蛋白质
免疫组化
Annexin A2
E6-associated protein
Transplanted tumor in nude mice
Blotting
Western
Immunohistochemisty
