WISP-2基因敲降的乳腺癌移植瘤生长相关分子机制研究  

A study on molecular mechanisms of WISP-2 knockdown for the growth of breast cancer xenografts

作　　者：吕艳[1] 张畅[4] 李小江[2] 郑旭[1] 韩燕燕[1] 高珊 李翀[1] 耿强[1] 兰福 赵杰[1] LYU Yan;ZHANG Chang;LI Xiaojiang(Department of Pathology,First Affiliated Hospital,Tianjin University of Traditional Chinese Medicine,Tianjin 300381,CHINA)

机构地区：[1]天津中医药大学第一附属医院病理科,300381 [2]天津中医药大学第一附属医院肿瘤科,300381 [3]天津中医药大学第一附属医院实验中心,300381 [4]天津中医药大学

出　　处：《江苏医药》2020年第5期469-472,F0002,共5页Jiangsu Medical Journal

基　　金：天津市教委科研计划项目(2017KJ160)。

摘　　要：目的观察Wnt-1诱导分泌蛋白2(WISP-2)基因敲降与乳腺癌细胞增殖活性的相关性以及WISP-2调控S期激酶相关蛋白2(Skp2)和p27Kip1在乳腺癌移植瘤形成中的作用。方法MCF-7乳腺癌细胞分为三组:A组为正常培养细胞;B组为WISP-2短发夹RNA(shRNA)质粒转染细胞;C组为空质粒转染细胞。MTT法检测WISP-2基因敲降对MCF-7细胞增殖活性的影响。取45只SPF级健康BALB/c雌性裸鼠,随机均分为D、E和F三组,分别于裸鼠右腋部皮下接种0.2 mL携带WISP-2 shRNA质粒转染、空质粒转染和正常培养的生长密度为每毫升1×10~6个的MCF-7细胞;14 d后,肿瘤最小直径达到0.5 cm视为移植瘤形成。分别采用荧光定量RT-PCR法和Western blot法检测三组移植瘤中WISP-2、Skp2和p27Kip1 mRNA和蛋白水平。结果B组细胞增殖活性高于A组和C组(P<0.05)。D组WISP-2和p27Kip1 mRNA和蛋白水平低于E组和F组(P<0.05),而Skp2 mRNA和蛋白水平高于E组和F组(P<0.05)。结论WISP-2基因敲降导致Skp2和p27Kip1 mRNA和蛋白水平异常,诱导乳腺癌细胞增殖,促进乳腺癌移植瘤形成。Objective To observe the correlation between Wnt-1 inducible secreted protein 2(WISP-2)gene knockdown and breast cancer cell proliferation and the role of WISP-2 in regulating S-phase kinase-associated protein 2(Skp2)and p27 Kip1 in the formation of breast cancer xenografts.Methods MCF-7 breast cancer cells were divided into three groups of A(normally cultured MCF-7 cells),B(WISP-2 shRNA plasmid transfected MCF-7 cells)and C(empty plasmid transfected MCF-7 cells).MTT assay was used to detect the effect of WISP-2 knockdown on MCF-7 cell proliferation activity.Forty-five SPF-grade healthy BALB/c female nude mice were randomly divided into three groups of D,E and F,which were inoculated subcutaneously 0.2 ml of WISP-2 shRNA plasmid transfected MCF-7 cells,empty plasmid transfected MCF-7 cells and normal cultured MCF-7 cells in the right armpit,respectively.After 14 days,the minimum tumor diameter of 0.5 cm was regarded as xenograft formation.The mRNA and protein levels of WISP-2,Skp2 and p27 Kip1 in the xenografts of three groups were detected by fluorescent quantitative RT-PCR and Western blot,respectively.ResultsThe cell proliferation activity in group B was higher than that in groups of A and C(P<0.05).The WISP-2 and p27 Kip1 mRNA and protein levels in group D were lower,while the mRNA and protein levels of Skp2 were higher than those in groups of E and F(P<0.05).Conclusion WISP-2 knockdown causes abnormal mRNA and protein expressions of Skp2 and p27 Kip1,induces breast cancer cell proliferation,and promotes breast cancer transplantation.

关 键 词：乳腺癌 Wnt-1诱导分泌蛋白2 短发夹核糖核酸 S期激酶相关蛋白2 P27KIP1 

分 类 号：R737[医药卫生—肿瘤]