摘要
目的探讨CCN5基因敲降对人乳腺癌细胞增殖的影响;探索CCN5在人乳腺癌细胞增殖中的作用机制;探寻CCN5与乳腺癌发生、发展中的相关性。方法设置CCN5siRNA转染组,慢病毒空载体转染组,空白对照组,分别采用不同处理方法。CCN5siRNA转染组(以下简称“转染组”):慢病毒载体介导的CCN5siRNA转染人乳腺癌细胞MCF-7;慢病毒空载体转染组(以下简称“空载组”):慢病毒空载体转染人乳腺癌细胞MCF-7;空白对照组不进行任何转染操作。采用MTT法检测各组细胞的增殖能力变化,采用RT-PCR、Western blot检测各组细胞CCN5、Skp2和P27Kip1mRNA及蛋白表达。结果MTT实验表明:转染组细胞增殖力高于空载组和空白对照组;RT-PCR和Western blot结果表明:转染组CCN5mRNA和蛋白水平低于空载组和空白对照组,差异有统计学意义(P<0.05);转染组Skp2、P27Kip1mRNA和蛋白水平高于空载组和空白对照组,差异有统计学意义(P<0.05)。结论CCN5siRNA抑制CCN5mRNA和蛋白表达,促进乳腺癌细胞增殖,并且通过上调Skp2、下调P27Kip1诱导乳腺癌细胞增殖。
Objective To investigate the effect of CCN5 gene knockdown on the proliferation of human breast cancer cells,to explore the mechanism of CCN5 in the proliferation of these cancer cells,and to explore the association of CCN5 with the occurrence and development of breast cancer.Methods CCN5siRNA transfection group,lentivirus empty vector transfection group and blank control group were set up by different treatment methods.CCN5siRNA transfection group(hereinafter referred to as transfection group):transfection of human breast cancer cells MCF-7 with CCN5siRNA mediated by lentiviral vector,lentivirus empty vector transfection group(hereinafter referred to as no-load group):transfection of cells MCF-7 with empty lentivirus vector.No transfection was performed in the blank control group.MTT assay was used to detect the changes in cell proliferation capacity in each group,so were RT-PCR and Western blotting to detect the mRNA and protein expressions of CCN5,Skp2 and p27Kip1.Results MTT assay showed that proliferation ability of MCF-7 in transfection group was higher than that in empty vector group and control group.The results of RT-PCR and Western blotting showed that the mRNA and protein levels of CCN5 in transfection group were lower than those in empty vector group and control group(P<0.05),the mRNA and protein levels of Skp2 in transfection group were higher than those in empty vector group and control group(P<0.05),while the mRNA and protein levels of p27Kip1 in transfection group were lower than those in empty vector group and control group(P<0.05).Conclusions CCN5siRNA inhibits the mRNA and protein expression of CCN5,promotes and induces the proliferation of breast cancer cells by up-regulating Skp2 and down-regulating P27Kip1.
作者
吕艳
郑旭
韩燕燕
高珊
李翀
耿强
LV Yan;ZHENG Xu;HAN Yanyan;GAO Shan;LI Chong;GENG Qiang(The First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300381,China)
出处
《实用医学杂志》
CAS
北大核心
2023年第22期2898-2902,共5页
The Journal of Practical Medicine
基金
天津市教委科研项目(编号:2017KJ160)。
