Sun Jun, Zhang Xiaowen, Chen Zimin, Ye Xiaoshuo, Zhang Chen
Division II of General Surgery, Department of Pediatric Surgery, Shenzhen Children's Hospital, Shenzhen, China.
Transl Cancer Res. 2024 Jul 31;13(7):3397-3406. doi: 10.21037/tcr-24-25. Epub 2024 Jul 16.
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.
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 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.
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.
MiR-1271-5p can promote the growth and migration of tumor cells by inhibiting ACY-1 expression in NB.
已发现氨基酰化酶1(ACY-1)是神经母细胞瘤(NB)中的一种肿瘤抑制基因。本研究旨在通过数据库预测分析鉴定并验证可能调控ACY-1的微小RNA(miRNA),并通过细胞实验验证miRNA与ACY-1在NB中的相互调控作用。
通过TargetScan、miRTarBase和其他四个数据库预测并筛选可能与ACY-1结合的miRNA,采用逆转录-定量聚合酶链反应(RT-qPCR)检测四组临床样本中预测的miRNA和ACY-1的表达,并筛选差异表达的miRNA。然后,根据基因序列构建荧光素酶载体,检测ACY-1是否与所选miRNA结合。接着,沉默miR-1271-5p表达,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表达,探讨ACY-1在NB中的功能及miR-1271-5p的调控作用。
ACY-1在NB中表达下调,miR-1271-5p在NB中表达上调,且miR-1271-5p可靶向ACY-1。沉默miR-1271-5p表达可降低细胞活力并抑制肿瘤进展。干扰细胞中ACY-1表达后,细胞活力增强,凋亡显著减少,迁移和侵袭能力增强。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抑制剂恢复了ACY-1表达并改善了ACY-1功能。
在NB中,miR-1271-5p可通过抑制ACY-1表达促进肿瘤细胞的生长和迁移。