AKAP12 Endogenous Transcripts Suppress The Proliferation, Migration And Invasion Of Colorectal Cancer Cells By Directly Targeting oncomiR-183-5p
AKAP12 Endogenous Transcripts Suppress The Proliferation, Migration And Invasion Of Colorectal Cancer Cells By Directly Targeting oncomiR-183-5p
复制标题
AKAP12内源转录本通过直接靶向oncomiR-183-5p抑制结直肠癌细胞的增殖、迁移和侵袭
DOI：
10.2147/ott.s207600
复制
发表时间：
2019-01-01
影响因子：
4
通讯作者：
Guan, Ming
中科院分区：
文献类型：
--
作者：
Hu, Tingting;Wu, Xuan;Guan, Ming
目的恢复抑制基因产物丧失的功能已成为基因治疗领域的研究热点。AKAP 12，也称为Gravin/AKAP 250，是一种肿瘤抑制基因，其失调可能是癌症进展的原因。本研究旨在探讨AKAP 12 mRNA是否通过调控大肠癌（CRC）细胞中癌基因miRNA的表达而发挥抗癌作用。方法通过生物信息学分析预测AKAP 12的miRNAs，并通过双荧光素酶报告基因分析和RT-qPCR进行验证。通过miRseq在早期CRC肿瘤组织中验证了microRNA表达的改变。通过细胞计数试剂盒-8（CCK-8）测定来测量细胞增殖。细胞侵袭和迁移分别通过transwell和伤口愈合实验检测。进行体内实验以证实体外发现。结果在所有miRNA中，AKAP 12与miRNA-183- 5 p的表达呈负相关。荧光素酶分析显示AKAP 12直接靶向miR-183- 5 p。miRseq数据显示，miR-183在肿瘤发展的早期也失调，并且在晚期亚期II CRC患者中上调（P<0.01）。体外和体内的机制分析表明，在CRC细胞中，抗miR-183- 5 p抑制细胞增殖、迁移和侵袭，而miR-183- 5 p过表达导致相反的效果。结论oncomiR-183- 5 p可促进大肠癌细胞的增殖、迁移和侵袭。AKAP 12 miRNA结合元件（MRE）抑制miRNA-183- 5 p活性。因此，AKAP 12表达的任何变化都会影响miRNA-183- 5 p。这可能是除了调节肿瘤抑制基因的蛋白质介导之外的另一种抗肿瘤机制。
Purpose Restoring lost function to suppressor gene products has captured the interest of the research community in the field of gene therapy. AKAP12, also known as Gravin/AKAP250, is a tumor suppressor gene, and its deregulation may be responsible for cancer progression. The aim of this study was to investigate whether AKAP12 mRNA has an anti-cancer function by regulating onco-miRNA expression in colorectal cancer (CRC) cells. Methods miRNAs targeting AKAP12 were predicted by bioinformatics analysis and further confirmed by dual-luciferase reporter assays and RT-qPCR. The altered expression of microRNA was validated in early-stage CRC tumor tissues by miRseq. Cell proliferation was measured by Cell Counting Kit-8 (CCK-8) assay. Cell invasion and migration were detected by transwell and wound healing assays, respectively. In vivo experiments were conducted to confirm the in vitro findings. Results Among all miRNAs, reversed correlation between AKAP12 expression and miRNA-183-5p expression was most significant. Luciferase assays revealed that AKAP12 directly targeted miR-183-5p. The miRseq data showed that miR-183 was also dysregulated at the early stage of tumor development and upregulated in late sub-stage II CRC patients (P<0.01). Mechanistic analysis both in vitro and in vivo demonstrated that anti-miR-183-5p depressed cell proliferation, migration, and invasion in CRC cells while miR-183-5p overexpression resulted in opposite effects. Conclusion Our findings suggested that oncomiR-183-5p promoted the proliferation, migration, and invasion of CRC cells. AKAP12 miRNA-binding elements (MREs) suppressed miRNA-183-5p activities. Any change in expression of AKAP12 thus affected miRNA-183-5p. This may be another anti-tumor mechanism in addition to protein-mediation that regulates tumor suppressor genes.