摘要
目的:探究过表达长链非编码RNA(lncRNA)LINC00886通过调控微小RNA-451a(miR-451a)对乳腺癌(BC)MDA-MB-231细胞生物学行为的影响。方法:荧光定量PCR(qRT-PCR)检测人乳腺上皮细胞系(MCF-12A)和4种BC细胞系(HCC1937、SUM159、SK-BR-3和MDA-MB-231)中lncRNA LINC00886、miR-451a表达。将MDA-MB-231细胞分为对照组、LINC00886-NC组、LINC00886组、LINC00886+miR-NC组、LINC00886+miR-451a组。qRT-PCR法检测MDA-MB-231细胞中lncRNA LINC00886、miR-451a表达水平;克隆形成实验和EdU染色检测MDA-MB-231细胞增殖能力;流式细胞术、Transwell小室实验、划痕愈合实验分别用来检测MDA-MB-231细胞凋亡、侵袭、迁移;蛋白印迹法检测MDA-MB-231细胞中增殖细胞核抗原(PCNA)、Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)、活化的半胱氨酸天冬氨酸蛋白酶3(Cleaved caspase-3)、基质金属蛋白酶(MMP)-2、MMP-9蛋白表达;双荧光素酶报告基因检测lncRNA LINC00886与miR-451a的靶向关系。结果:与人乳腺上皮细胞MCF-12A相比,BC细胞系中lncRNA LINC00886表达水平降低,miR-451a表达水平升高(P<0.05);过表达lncRNA LINC00886可升高MDA-MB-231细胞凋亡率、Bax和Cleaved caspase-3蛋白表达,降低miR-451a表达、集落形成数、EdU阳性细胞百分比、侵袭细胞数、迁移率以及PCNA、Bcl-2、MMP-2、MMP-9蛋白表达(P<0.05);上调miR-451a表达可减弱lncRNA LINC00886过表达对MDA-MB-231细胞的影响(P<0.05);lncRNA LINC00886可靶向调控miR-451a表达。结论:过表达lncRNA LINC00886可通过靶向抑制miR-451a表达促进MDA-MB-231细胞凋亡并抑制MDA-MB-231细胞增殖、侵袭和迁移。
Objective:To explore the effects of overexpression of long non-coding RNA(lncRNA)LINC00886 on the biological behavior of breast cancer(BC)MDA-MB-231 cells by regulating microRNA-451a(miR-451a).Methods:Fluorescence quantitative PCR(qRT-PCR)was used to measure the expression of lncRNA LINC00886 and miR-451a in human breast epithelial cell line(MCF-12A)and four BC cell lines(HCC1937,SUM159,SK-BR-3 and MDA-MB-231).MDA-MB-231 cells were divided into control group,LINC00886-NC group,LINC00886 group,LINC00886+miR-NC group and LINC00886+miR-451a group.The expression levels of lncRNA LINC00886 and miR-451a in MDA-MB-231 cells were detected by qRT-PCR.The proliferation ability of MDA-MB-231 cells was detected by clone formation experiment and EdU staining.The apoptosis,invasion and migration of MDA-MB-231 cells were detected by flow cytometry,Transwell chamber assay and scratch healing assay,respectively.The protein expression levels of proliferating cell nuclear antigen(PCNA),Bcl-2 associated X protein(Bax),B cell lymphocytoma-2(Bcl-2),activated cysteine aspartic protease 3(Cleaved caspase-3),matrix metalloproteinase(MMP)-2,MMP-9 of MDA-MB-231 cells were detected by Western blot.The targeting relationship between lncRNA LINC00886 and miR-451a was detected by dual luciferase reporter gene.Results:Compared with human breast epithelial cells MCF-12A,the expression level of lncRNA LINC00886 in BC cell lines was significantly decreased,and the expression level of miR-451a was significantly increased(P<0.05).The overexpression of lncRNA LINC00886 can increase the apoptosis rate of MDA-MB-231 cells,the expression of Bax and Cleaved caspase-3 protein,and reduce miR-451a expression,colony formation number,percentage of EdU positive cells,number of invasive cells,migration rate,protein expression of PCNA,Bcl-2,MMP-2,MMP-9(P<0.05).The up-regulation of miR-451a expression could attenuate the effect of overexpression of lncRNA LINC00886 on MDA-MB-231 cells(P<0.05).lncRNA LINC00886 can target the expression of miR-451a.Conclusion:Overexpression of lncRNA LINC00886 can promote the apoptosis of MDA-MB-231 cells and inhibit the proliferation,invasion and migration of MDA-MB-231 cells by targeting the inhibition of miR-451a expression.
作者
高建朝
张京力
张志生
李晓霞
马科
冯志林
周海丰
王展海
梁晚平
GAO Jianchao;ZHANG Jingli;ZHANG Zhisheng;LI Xiaoxia;MA Ke;FENG Zhilin;ZHOU Haifeng;WANG Zhanhai;LIANG Wanping(Department of Breast Surgery,the First Hospital Affiliated to Hebei North University,Hebei Zhangjiakou 075000,China.)
出处
《现代肿瘤医学》
CAS
北大核心
2023年第14期2593-2600,共8页
Journal of Modern Oncology
基金
河北省医学科学研究课题计划项目(编号:20200569)
张家口市重点研发计划项目(编号:2121135D)。
