神经母细胞瘤(NB)是外周交感神经系统的罕见儿童恶性肿瘤,约占儿科肿瘤的10%。microRNA(miR)-125a已被暗示在各种癌症中充当肿瘤抑制因子。在这里,我们着手确定miR-125a是否在NB中发挥抗肿瘤作用.
通过NB组织和癌旁组织的miRNA微阵列分析鉴定下调的miRNA。逆转录-定量(RT-q)PCR检测miR-125a在NB组织和细胞中的表达,然后进行预后分析。对差异表达的miRNA的靶基因进行基因本体论(GO)富集分析。细胞增殖,凋亡,细胞计数试剂盒-8(CCK-8)检测分化,Hoechst染色,免疫荧光,和westernblot.将NB细胞注射到裸鼠体内以检测致瘤性,凋亡,和体内分化活性。进行了双荧光素酶测定和染色质免疫沉淀(ChIP)以验证miR-125a与PHOX2B或组蛋白脱乙酰酶2(HDAC2)之间的结合关系,分别。最后,进行了救援实验。
miR-125a在NB组织和细胞中下调,这与不良预后有关。miR-125a降低NB细胞增殖并增强凋亡和分化。miR-125a过表达的NB细胞减少了细胞肿瘤发生,并增加了异种移植肿瘤组织中的凋亡和分化。miR-125a靶向PHOX2B,在NB组织和细胞中高表达。HDAC2,在NB组织和细胞中高表达,通过组蛋白去乙酰化抑制miR-125a转录。HDAC2或PHOX2B的过表达拯救了miR-125a对NB细胞增殖的影响,凋亡,和差异化。
HDAC2通过去乙酰化抑制miR-125a转录,miR-125a通过与PHOX2B结合抑制NB发育。
Neuroblastoma (NB) is an infrequent childhood malignancy of the peripheral sympathetic nervous system and is accountable for about 10% of pediatric tumors. microRNA (miR)-125a has been implicated to serve as a tumor suppressor in various cancers. Herein, we set out to ascertain whether miR-125a exerts antitumor effects in NB.
Downregulated miRNAs were identified by miRNA microarray analysis of NB tissues and paracancerous tissues. The expression of miR-125a in NB tissues and cells was detected by reverse transcription-quantitative (RT-q) PCR, followed by prognostic analysis. Gene Ontology (GO) enrichment analysis was performed on target genes of differentially expressed miRNAs. Cell proliferation, apoptosis, and differentiation were detected by cell counting kit-8 (CCK-8), Hoechst staining, immunofluorescence, and western blot. NB cells were injected into nude mice to detect tumorigenic, apoptotic, and differentiation activities in vivo. Dual-luciferase assay and chromatin immunoprecipitation (ChIP) were carried out to verify the binding relationship between miR-125a and
PHOX2B or histone deacetylases 2 (HDAC2), respectively. Finally, rescue experiments were conducted.
miR-125a was downregulated in NB tissues and cells, which was associated with poor prognosis. miR-125a reduced NB cell proliferation and augmented apoptosis and differentiation. NB cells with miR-125a overexpression decreased cell tumorigenesis and increased apoptosis and differentiation in xenograft tumor tissues. miR-125a targeted
PHOX2B, which was highly expressed in NB tissues and cells. HDAC2, highly expressed in NB tissues and cells, repressed miR-125a transcription through histone deacetylation. Overexpression of HDAC2 or
PHOX2B rescued the effects of miR-125a on NB cell proliferation, apoptosis, and differentiation.
HDAC2 inhibited miR-125a transcription through deacetylation, and miR-125a suppressed NB development through binding to
PHOX2B.