Long non-coding RNA CCAT1 promotes human retinoblastoma SO-RB50 and Y79 cells through negative regulation of miR-218-5p
| Autor: | Jianguang Zhong, Yongping Hu, Hongxu Zhang, Peng You, Zhenyu Bian, Xiang Fang |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
G2 Phase
0301 basic medicine Cell Apoptosis Biology Transfection Flow cytometry 03 medical and health sciences 0302 clinical medicine Cell Movement Cell Line Tumor medicine Humans RNA Small Interfering Cyclin B1 Cell Proliferation bcl-2-Associated X Protein Pharmacology Cyclin-dependent kinase 1 medicine.diagnostic_test Caspase 3 Cell growth Retinoblastoma General Medicine Cell cycle Molecular biology Caspase 9 Long non-coding RNA Gene Expression Regulation Neoplastic MicroRNAs 030104 developmental biology medicine.anatomical_structure Proto-Oncogene Proteins c-bcl-2 030220 oncology & carcinogenesis RNA Long Noncoding Cell Division |
| Zdroj: | Biomedicine & Pharmacotherapy. 87:683-691 |
| ISSN: | 0753-3322 |
| DOI: | 10.1016/j.biopha.2017.01.004 |
| Popis: | To investigate the regulatory role and potential mechanism of long non-coding RNAs (lncRNA) in human retinoblastoma (RB).The lncRNA profile in RB tissues were analyzed by microarray and quantitative reverse transcription PCR (qRT-PCR). One of the identified lncRNAs (LncRNA CCAT1) was selected for further experiments. SO-RB50 and Y79 cells were transfected with negative control, siRNA targeting lncRNA CCAT1 (si-CCAT1) and si-CCAT1+miR218-5p inhibitor, respectively. lncRNA CCAT1 expression was measured by qRT-PCR. Cell proliferation, migration and invasion were detected by CCK8, wound scratching, and transwell assay, respectively. Apoptosis and cell cycle distribution were assessed by flow cytometry. Apoptosis- (cle-caspase-3, cle-caspase-9, Bax and Bcl-2) and cell cycle-related protein expression (cyclin B1, CDC2 and p-CDC2 (Thr161)) were analyzed by Western blot.lncRNA CCAT1 expression in SO-RB50 and Y79 cells was significantly inhibited after si-CCAT1 transfection (P0.01). Both RB cells exhibited significantly reduced proliferation, migration and invasion abilities, but markedly increased apoptosis at 48h after si-CCAT1 transfection (P0.05 or 0.01). RB cells in si-CCAT1+miR218-5p inhibitor group had significantly higher proliferation, migration and invasion, but notably lower apoptosis compared with si-CCAT1 group at 24 and 48h after transfection (all P0.05 or 0.01). si-CCAT1 significantly increased the expression of cle-caspase-3, cle-caspase-9, Bax, but decreased Bcl-2 expression (P0.01). The proportion of G2/M SO-RB50 and Y79 cells in siCCAT1 group was significantly increased compared with negative control group (P0.01). LncRNA CCAT1 interference significantly reduced the expression of cyclin B1, CDC2 and p-CDC2 (Thr161) (P0.01).LncRNA CCAT1 promotes the proliferation migration and invasion, and reduces cell apoptosis of SO-RB50 and Y79 cells, probably through negative modulation of miR-218-5p. Our study suggested lncRNA CCAT1 as a potential biomarker and therapeutic target for RB. |
| Databáze: | OpenAIRE |
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