MicroRNA‑125 inhibits RKO colorectal cancer cell growth by targeting VEGF
| Autor: | Jia‑Wei Zhu, Hua‑Qing Zhu, He‑Yang You, Xin Yin, Yi‑Rong Lin, Jing Chen, Qi‑Bing Wu |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Vascular Endothelial Growth Factor A
0301 basic medicine MAP Kinase Signaling System Cell Apoptosis 03 medical and health sciences 0302 clinical medicine Western blot Cell Movement Cell Line Tumor Genetics medicine Humans MTT assay Viability assay Luciferases 3' Untranslated Regions Cell Proliferation Base Sequence Cell Death Oncogene medicine.diagnostic_test Chemistry Cell growth General Medicine Transfection Cell cycle Molecular biology Gene Expression Regulation Neoplastic MicroRNAs 030104 developmental biology medicine.anatomical_structure Cyclooxygenase 2 030220 oncology & carcinogenesis Colorectal Neoplasms |
| Zdroj: | International Journal of Molecular Medicine. |
| ISSN: | 1791-244X 1107-3756 |
| DOI: | 10.3892/ijmm.2018.3632 |
| Popis: | Colorectal cancer (CRC) is one of the major types of cancer and causes of mortality worldwide, and it remains the third most common cause of cancer‑associated mortality worldwide. MicroRNAs (miRNAs) are a class of small RNAs, which have been shown to be associated with CRC. In the present study, an MTT assay and proliferating cell nuclear antigen (PCNA) protein examination assay were performed to detect RKO cell viability. Hoechst staining, and caspase‑3 activity and BrdU incorporation assays were performed to detect RKO cell apoptosis, respectively. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analyses were used to analyze the expression of cyclooxygenase‑2 (COX‑2). Western blot analysis was also used to analyze the expression of vascular endothelial growth factor (VEGF) and mitogen‑activated protein kinase (MAPK) signal molecules, including extracellular signal‑regulated kinase (ERK), p38 and c‑Jun N‑terminal kinase (JNK). The target genes of miR-125 were predicted using a double luciferase reporter gene assay. The results of the MTT assay showed that RKO cell viability was decreased by an miRNA-125 mimic and increased by the miRNA-125 inhibitor. The RKO cell viability was significantly correlated with the expression of PCNA. The migration of RKO cells was significantly downregulated in the miR-125 mimics‑transfected cells and upregulated in the miRNA-125 inhibitor‑transfected cells. The results of Hoechst staining and the caspase‑3 activity and BrdU incorporation assays showed that RKO cell apoptosis was increased following miRNA-125 mimic transfection and decreased following miRNA-125 inhibitor transfection. The results of the RT‑qPCR and western blot analysis showed that the expression of COX‑2 was increased in the miR-125 mimic‑transfected cells and decreased in the miR-125 inhibitor‑transfected cells. Using an online miRNA target prediction database, the double luciferase reporter gene assay showed that miR‑125 targeted and inhibited the expression of VEGF through target sites located in the 3' untranslated region of VEGF mRNA. In conclusion, the abnormal expression of miR‑125 was found to be closely associated with CRC. Therefore, miR‑125 may be a novel therapeutic target for CRC. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|