■氨基酰基转移酶1(ACY-1)已被发现是神经母细胞瘤(NB)中的抑癌基因。本研究旨在通过数据库预测分析鉴定和验证可能调控ACY-1的microRNAs(miRNAs),并通过细胞实验验证miRNA和ACY-1在NB中的相互调节作用。
■通过TargetScan预测并选择了可能结合ACY-1的miRNA,miRTarBase和其他四个数据库,通过逆转录-定量聚合酶链反应(RT-qPCR)检测四组临床样本中预测的miRNAs和ACY-1的表达,并筛选差异表达的miRNA。然后,根据ACY-1基因序列构建荧光素酶载体以检测ACY-1是否与所选miRNA结合。然后,沉默miR-1271-5p表达以检测miR-1271-5p在NB生长和迁移中的功能。最后,沉默ACY-1和miR-1271-5p以改变ACY-1的表达,并探讨了ACY-1在NB中的功能和miR-1271-5p的调控作用。
■ACY-1在NB中下调,miR-1271-5p在NB中上调,miR-1271-5p可以靶向ACY-1。沉默miR-1271-5p表达可以降低细胞活力并抑制肿瘤进展。在干扰细胞中ACY-1表达后,细胞活力增强,细胞凋亡显著减少,移民和入侵得到加强。部分恢复ACY-1表达后,si-ACY-1对细胞的作用减弱。在SK-N-SH和SH-SY-5Y细胞中,miR-1271-5p抑制剂可恢复ACY-1的表达并改善ACY-1的功能.
■MiR-1271-5p可以通过抑制NB中ACY-1的表达来促进肿瘤细胞的生长和迁移。
UNASSIGNED: Aminoacylase 1 (ACY-1) has been found to be a tumor suppressor gene in neuroblastoma (NB). This study aimed to identify and verify the microRNAs (miRNAs) that may regulate ACY-1 through database prediction analysis, and verify the mutual regulatory effect of miRNA and ACY-1 in NB through cell experiments.
UNASSIGNED: The miRNAs that might bind ACY-1 were predicted and selected by TargetScan, miRTarBase and four other databases, the expression of the predicted miRNAs and ACY-1 was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) in four groups of clinical samples, and the differentially expressed miRNAs were screened. Then, luciferase vector was constructed according to the ACY-1 gene sequence to detect whether ACY-1 binds to the selected miRNA. Then, miR-1271-5p expression was silenced to detect miR-1271-5p function in the growth and migration of NB. Finally, ACY-1 and miR-1271-5p were silenced to change ACY-1 expression, and ACY-1 function in NB and the regulatory role of miR-1271-5p were explored.
UNASSIGNED: ACY-1 was downregulated in NB, miR-1271-5p was upregulated in NB, and miR-1271-5p could be targeted to ACY-1. Silencing miR-1271-5p expression can reduce cell viability and inhibit tumor progression. After interfering with ACY-1 expression in cells, cell viability was enhanced, apoptosis was significantly decreased, and migration and invasion were enhanced. After partially restoring ACY-1 expression, the effect of si-ACY-1 on cells was weakened. In SK-N-SH and SH-SY-5Y cells, the miR-1271-5p inhibitor restored ACY-1 expression and improved ACY-1 function.
UNASSIGNED: MiR-1271-5p can promote the growth and migration of tumor cells by inhibiting ACY-1 expression in NB.