摘要
目的观察Kruppel样因子15(KLF15)基因在乳腺癌组织中的表达变化及对乳腺癌细胞MCF-7增殖能力的影响,并探讨相关机制。方法分别利用Cancer RNA-seq Nexus(CRN,http://syslab4.nchu.edu.tw/)和UALCAN(http://ualcan.path.uab.edu/index.html)分析TCGA数据库中乳腺癌样本的RNA序列数据,分析乳腺癌组织中KLF15 mRNA的表达变化。将MCF-7细胞分为过表达组、空载组,过表达组转染pcDNA3.1-KLF15过表达质粒,空载组转染pcDNA3.1空载质粒。采用MTT法检测过表达组和空载组细胞增殖能力,采用集落形成实验测算细胞克隆形成率,流式细胞术检测细胞周期,采用RT-qPCR法检测p21、p27基因,Western blotting法检测p21、p27蛋白。结果分析TCGA数据库中乳腺癌的RNA序列数据显示,乳腺癌组织中KLF15 mRNA表达低于正常乳腺组织(P<0.05)。过表达组培养24、48 h细胞增殖能力均低于空载组(P均<0.05)。过表达组克隆形成率低于空载组(P<0.05)。过表达组G0/G1期细胞比例高于空载组(P<0.05)。过表达组p21基因及蛋白表达高于空载组(P均<0.05)。结论KLF15基因在乳腺癌组织中低表达。KLF15基因表达上调的乳腺癌细胞增殖能力、克隆形成能力减弱,细胞周期阻滞。KLF15对乳腺癌细胞的增殖抑制机制可能与上调p21表达有关。
Objective To observe the expression changes of Kruppel-like factor 15(KLF15)gene in breast cancer tissues and to explore the underling mechanism of its effect on the proliferation in MCF-7 cells.Methods RNA sequence data of breast cancer samples in TCGA database was analyzed by Cancer RNA-seq Nexus(CRN,http://syslab4.nchu.edu.tw/)and UALCAN(http://manualcan.path.uab.edu/index.html),and then we analyzed the expression changes of KLF15 mRNA in the breast cancer tissues.MCF-7 cells were divided into the overexpression group and empty group.pcDNA3.1-KLF15 overexpression plasmid was transfected into overexpression group,and pcDNA3.1 empty plasmid was transfected into the empty group.The proliferation ability of MCF-7 cells was observed by MTT assay,the cloning efficiency was calculated by colony formation experiment,the cell cycle distribution was analyzed using flow cytometry,and the mRNA and protein expression of p21 and p27 was detected by RT-qPCR and Western blotting.Results Using CRN and UALCAN to analyze the RNA sequence data of breast cancer in TCGA database,we found the expression of KLF15 mRNA in the breast cancer tissues was lower than that of the normal breast tissues(P<0.05).The proliferation abilities of the overexpression group at 24 and 48 h was lower than those of the empty group(both P<0.05).The clone formation rate of the overexpression group was also lower than that of the empty group(P<0.05).The percentage of cells in the G 0/G 1 phase in the overexpression group was higher than that of the empty group(P<0.05).p21 mRNA and protein expression in the overexpression group was higher than that of the empty group(P<0.05).Conclusions KLF15 is low expressed in the breast cancer tissues.Up-regulated expression of KLF15 can decrease the proliferation ability and clone formation ability of breast cancer cells,it can also lead to cell cycle arrest.The inhibition of KLF15 on breast cancer cell proliferation may be related to the up-regulation of p21 expression.
作者
高温杰
王振国
刘显胜
任贵兵
GAO Wenjie;WANG Zhenguo;LIU Xiansheng;REN Guibing(Logistics University of People's Armed Police Force,Tianjin 300300,China)
出处
《山东医药》
CAS
2019年第32期25-28,共4页
Shandong Medical Journal
基金
天津市科技计划项目(15ZXLCSY00040)
