OncoTargets and Therapy (Oct 2019)
AKAP12 Endogenous Transcripts Suppress The Proliferation, Migration And Invasion Of Colorectal Cancer Cells By Directly Targeting oncomiR-183-5p
Abstract
Tingting Hu,1,* Xuan Wu,2,* Ke Li,2 Yuan Li,2 Ping He,3 Zhiyuan Wu,4 Jie Fan,5 Weiwei Liu,2,6 Ming Guan1,4 1Department of Clinical Laboratory, Huashan Hospital, Fudan University, Shanghai, People’s Republic of China; 2Central Laboratory and Department of Laboratory Medicine, Shanghai Tenth People’s Hospital, Tongji University, Shanghai, People’s Republic of China; 3Department of Nuclear Medicine, Nanjing Hospital, Nanjing Medical University, Nanjing, People’s Republic of China; 4Department of Clinical Laboratory, Huashan Hospital North, Fudan University, Shanghai, People’s Republic of China; 5Department of Pathology, Huashan Hospital, Fudan University, Shanghai, People’s Republic of China; 6Department of Laboratory Medicine, Shanghai Skin Disease Hospital, Tongji University, Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Weiwei LiuCentral Laboratory and Department of Laboratory Medicine, Shanghai Tenth People’s Hospital, Tongji University, No. 301 Middle Yanchang Road, Shanghai, People’s Republic of ChinaTel/fax +86-21-6630 6905Email [email protected] GuanDepartment of Clinical Laboratory, Huashan Hospital, Fudan University, No. 12 Middle Wulumuqi Road, Shanghai, People’s Republic of ChinaTel/fax +86-21-5288 8048Email [email protected]: 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.Keywords: microRNA, cell proliferation, cell migration, cell invasion, AKAP12 endogenous transcripts, colorectal cancer cells, oncomiR-183-5p
