摘要
目的 研究着丝粒关联蛋白1(KNTC1)在乳腺癌组织和细胞中的表达、功能及其发挥功能的潜在机制。方法 下载癌症基因组图谱乳腺癌中的RNA-seq数据,一般线性模型估算KNTC1在不同组间的差异。qRT-PCR检测人乳腺癌细胞MDA-MB-231、T-47D、MCF-7中KNTC1的mRNA表达水平。使用含有目的基因RNA干扰序列的慢病毒技术敲减MDA-MB-231细胞中KNTC1表达,用qRT-PCR和Western blot检测细胞中KNTC1 mRNA和蛋白质表达水平。Celigo细胞计数和克隆形成试验检测细胞生长和增殖状况;流式细胞术检测细胞凋亡率。结果 KNTC1在乳腺癌组织中高度表达。KNTC1在MDA-MB-231、T-47D和MCF-7细胞中的表达丰度均为高,在MDA-MB-231细胞表达最高。与对照组比较,实验组MDA-MB-231细胞中KNTC1表达后,细胞生长与增殖能力显著下降(7.82±0.73、0.89±0.04;t=17.259,P=0.003),细胞的克隆形成能力显著降低(75±2、5±2;t=121.244,P<0.001),细胞凋亡明显增加[(4.73±0.19)%、(8.86±0.12)%;t=31.506,P=0.001]。结论 KNTC1在乳腺癌组织和细胞中高度表达,干扰KNTC1表达可抑制乳腺癌细胞的增殖克隆并促进其凋亡。
Objective To investigate the expression and function of kinetochore-associated protein 1(KNTC1)in breast cancer,and to understand the potential mechanisms by which it exerts its function.Methods RNA-seq data of breast cancer of The Cancer Genome Atlas were downloaded.General linear model was used to estimate the differential expression of KNTC1 among different subgroups.The mRNA expression levels of KNTC1 in human breast cancer cell MDA-MB-231,T-47D,and MCF-7 were detected by qRT-PCR.KNTC1 expression was knocked down in MDA-MB-231 cells by using a lentiviral technique containing RNA interference sequences of the target gene.QRT-PCR and Western blot were used to detect KNTC1 mRNA and protein expression levels in the cells.Celigo cell counting and colony-forming assay were performed to detect cell growth and proliferation status,and flow cytometry to detect apoptosis rate.Results KNTC1 was highly expressed in breast cancer tissues.KNTC1 was highly expressed in MDA-MB-231,T-47D,and MCF-7 cells,and the highest expression was in MDA-MB-231 cells.Compared with the control group,in the MDA-MB-231 cells of the observation group,cell growth and proliferation capacity was obviously decreased(7.82±0.73 and 0.89±0.04;t=17.259,P=0.003),the clonogenic ability of cells was apparently reduced(75±2 and 5±2;t=121.244,P<0.001),cell apoptosis was significantly increased[(4.73±0.19)%and(8.86±0.12)%;t=31.506,P=0.001].Conclusion KNTC1 is highly expressed in breast cancer tissue and human breast cancer cell,not only interfering KNTC1 expression inhibits proliferation and clone of MDA-MB-231 cells,but promotes apoptosis of breast cancer cells.
作者
安天棋
宗红
赵瑞华
AN Tianqi;ZONG Hong;ZHAO Ruihua(Department of Oncology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)
出处
《肿瘤基础与临床》
2023年第2期93-98,共6页
journal of basic and clinical oncology
基金
河南省自然科学基金资助项目(202300410383)。
关键词
着丝粒关联蛋白1
乳腺癌
细胞增殖
靶向治疗
kinetochore-associated protein 1
breast cancer
cell proliferation
targeted therapy
