Lnc-FAM83D-3通过miR-504-3p-FAM83D轴调控三阴性乳腺癌的机制研究

Effect of Lnc-FAM83D-3 on triple negative breast cancer cells through targeting miR-504-3p-FAM83D axis

目的探讨长链非编码RNA FAM83D-3(lnc-FAM83D-3)在三阴性乳腺癌(TNBC)中的表达及其对HCC1937、MDA-MB-231生物学行为的影响及作用机制。方法通过TCGA数据库分析lnc-FAM83D-3在乳腺癌中的表达情况及病理相关性。RT-qPCR检测lnc-FAM83D-3和微小RNA-504-3p(miR-504-3p)在组织标本和乳腺癌细胞系中的表达;慢病毒转染干扰lnc-FAM83D-3表达,利用MTT、细胞划痕实验、侵袭实验和流式细胞术检测实验组与对照组细胞功能改变。使用生物信息学软件和双荧光素酶报告基因实验分析lnc-FAM83D-3和miR-504-3p及FAM83D之间的关系。FISH实验证实lnc-FAM83D-3在胞浆内的表达,Western bolt检测下游蛋白变化情况。结果在乳腺癌组织、癌细胞HCC1937和MDA-MB-231中lnc-FAM83D-3的表达增加(P<0.01)。与对照组相比,干扰lnc-FAM83D-3表达后,TNBC细胞株增殖、迁移和侵袭能力降低,凋亡比例增加;miR-504-3p表达上升,FAM83D表达下降;双荧光素酶报告基因实验结果显示lnc-FAM83D-3靶向调控miR-504-3p表达,miR-504-3p进一步负向调控FAM83D的水平。结论Lnc-FAM83D-3在TNBC细胞株中表达增加且通过抑制miR-504-3p促进FAM83D表达,影响乳腺癌恶性生物学行为。

Objective To investigate the expression of long noncoding-FAM83 D-3 in triple negative breast cancer(TNBC)cells,and its effects on proliferation,migration,invasion and apoptosis of HCC1937 and MDA-MB-231 cells.Methods The expression and pathological correlation of lnc-FAM83 D-3 in TNBC cells was analyzed by TCGA database.The expression levels of lnc-FAM83 D-3 and miR-504-3 p were assessed by RT-qPCR.Following down-regulation of lnc-FAM83 D-3 with lentivirus,proliferation,migration and apoptosis of cells were measuredusing methyl thiazolyltetrazolium,matrigel invasion assay and flow cytometry,respectively.The relationships within lnc-FAM83 D-3,miR-504-3 p and FAM83 D were analyzed by bioinformatics software and luciferase reporter gene assay.The location of lnc-FAM83 D-3 was detected by FISH assay.The level of FAM83 D was measured by western blotting.Results In HCC1937 and MDA-MB-231 cells,the expression levels of lnc-FAM83 D-3 was upregulated(P<0.01).The expression levels of miR-504-3 p in shlncRNA were higher than those in the control group,while the expression levels of FAM83 D protein were lower.Down-regulation of lnc-FAM83 D-3 significantly inhibited proliferation,migration and invasion of HCC1937 and MDA-MB-231 cells.Luciferase reporter gene assay showed that lnc-AM83 D-3 directly interacted with miR-504-3 p and down-regulated its expression,while miR-504-3 p negatively regulated the level of FAM83 D.Conclusions Expression of lnc-FAM83 D-3 is upregulated in HCC1937 and MDA-MB-231 cells.Lnc-FAM83 D-3 stimulates proliferation,migration and inhibits apoptosis of HCC1937 and MDA-MB-231 cells by regulating miR-504-3 p/FAM83 D axis.