Functional classification of prostate cancer‑associated miRNAs through CRISPR/Cas9‑mediated gene knockout
| Autor: | Weide Zhong, Yang Yang, Jianguo Zhu, Yuxiang Liang, Yue ping Wan, Guan‑Xing Chen, Cui‑Yun Zou, Zhao‑Dong Han, Fu‑Neng Jiang, Ze‑Zhen Liu, Wang Wei |
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| Rok vydání: | 2020 |
| Předmět: |
Male
0301 basic medicine Cancer Research microRNA-1225-5p microRNA-663a Biology Biochemistry Gene Knockout Techniques 03 medical and health sciences Prostate cancer 0302 clinical medicine Cell Movement Cell Line Tumor microRNA LNCaP Genetics medicine Humans CRISPR CRISPR/Cas9 Molecular Biology Gene knockout Cell Proliferation Regulation of gene expression Oncogene Prostatic Neoplasms Articles prostate cancer medicine.disease Gene Expression Regulation Neoplastic MicroRNAs 030104 developmental biology Oncology Tumor progression 030220 oncology & carcinogenesis Disease Progression Cancer research Molecular Medicine CRISPR-Cas Systems Glycolysis |
| Zdroj: | Molecular Medicine Reports |
| ISSN: | 1791-3004 1791-2997 |
| Popis: | The aim of the present study was to use the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR‑associated (Cas) 9‑mediated gene knockout technology for the rapid classification of the differential function of micro (mi)RNAs screened using miRNA expression profiling by microarray. The rational design of single guide RNAs for the CRISPR/Cas9 system was verified to function in human LNCaP cells with rapid and efficient target gene editing. miRNA (miR)‑205, miR‑221, miR‑222, miR‑30c, miR‑224, miR‑455‑3p, miR‑23b and miR‑505 were downregulated in patients with prostate cancer (PCa) and were experimentally validated to function as tumor suppressors in prostate cancer cells, affecting tumor proliferation, invasion and aerobic glycolysis. In addition, the data of the present study suggested that miR‑663a and mfiR‑1225‑5p were upregulated in prostate cancer tissues and cell proliferation of miR‑663a and miR‑1225‑5p knockout PCa cells was significantly lower compared with miR‑NC cells. Furthermore, knockout of miR‑1225‑5p and miR‑663a significantly decreased the lactate production in LNCaP cells in vitro. In conclusion, the present study offered a simple and efficient method for rapidly classifying miRNA function by applying CRISPR/Cas9 in LNCaP cells. The present study suggested, for the first time to the best of the authors' knowledge, that the aberrant expression of miR‑663a and miR‑1225‑5p may be involved with the progression of prostate cancer, implying their potential as candidate markers for this type of cancer. However, the precise role of miR‑663a and miR‑1225‑5p in accelerating the development of prostate cancer and promoting tumor progression remains to be elucidated. |
| Databáze: | OpenAIRE |
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