摘要
目的探讨shRNA靶向沉默接触蛋白-1(CNTN1)基因表达对人乳腺癌细胞株MDA-MB-468裸鼠皮下移植瘤生长的抑制作用。方法18只4周龄雄性BALB/c裸鼠随机均分为3组,收集处于对数生长期的MDA-MB-468细胞(空白对照组)、转染无义shRNA片段的MDA-MB-468细胞(空载体组)及转染shRNA载体片段的MDA-MB-468细胞(沉默组)分别于每组裸鼠后背部皮下注射制备裸鼠皮下移植瘤模型,观察沉默CNTN1基因对MDA-MB-468细胞移植瘤生长的影响以及采用免疫组织化学方法检测移植瘤中CNTN1与Ki-67蛋白的表达情况。结果各组裸鼠皮下接种肿瘤细胞后均成功致瘤,成瘤率为100%。每3d测量种植瘤体积发现沉默组种植瘤生长速度显著低于空白对照组(P<0.05)和空载体组(P<0.05),至第18天处死裸鼠获取肿瘤,发现沉默组种植瘤的体积及质量均显著小或低于空白对照组(P<0.05)和空载体组(P<0.05);沉默组种植瘤中CNTN1和Ki-67蛋白表达阳性率均明显低于空白对照组(P<0.05)和空载体组(P<0.05)。结论沉默CNTN1基因表达可以抑制人乳腺癌细胞株MDA-MB-468细胞裸鼠移植瘤的生长。
Objective To investigate the inhibitory effect of short hairpin RNA(shRNA)mediated contactin-1(CNTN1)gene silencing on growth of human breast cancer cell line MDA-MB-468 transplanted tumors in nude mice.Methods Eighteen nude mice(4-week-old male BALB/c)were randomly equally divided into three groups:blank control group,empty vector group,and silencing group.The MDA-MB-468 cells(blank control group),MDA-MB-468 cells transfected by nonsense shRNA(empty vector group),and MDA-MB-468 cells transfected by shRNA(silencing group)were collected in the logarithmic growth period,respectively.The subcutaneous tum or m odels of nude mice were prepared by the subcutaneous injection of the different group cells.The tumor growth was observed and the expressions of CNTN1 and Ki-67 proteins in the transplanted tumor were detected by the im m unohistochem istry.Results The xenograft models of human breast cancer cells were established successfully.The tumor growth in the silencing group was significantly slower than that of the other two groups at every 3 d point(P<0.05).The tumor volume and the tumor weight in the silencing group were significantly smaller or slighter than those of the other two groups at day 18(P<0.05).The positive rates of CNTN1 and Ki-67 protein expressions in the tumor tissues of the silencing group were lower than those of the other two groups(P<0.05),respectively.Conclusion Silencing expression of CNTN1 gene might inhibit growth of breast cancer cell line MDA-MB-468 transplanted tumors in mude mice.
作者
贺赛
耿洁
杨晓民
王虎霞
韩丕华
侯艳妮
范拥国
陈楠
HE Sai;GENG Jie;YANG Xiaomin;WANG Huxia;HAN Pihua;HOU Yanni;FAN Yongguo;CHEN Nan(Department of Breast Oncology,Shaanxi Provincial Cancer Hospital,Xi'an 710061,P.R.China;Department of Cardiovascular Internal Medicine,The Second Affiliated Hospital of Medical College of Xi'an Jiaotong University,Xi’an 710004,P.R.China)
出处
《中国普外基础与临床杂志》
CAS
2020年第11期1371-1374,共4页
Chinese Journal of Bases and Clinics In General Surgery
