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Detection of a Circulating MicroRNA Custom Panel in Patients with Metastatic Colorectal Cancer
Published on: March 14, 2019
Potential using of microRNA-34A in combination with paclitaxel in colorectal cancer cells
Hadis Soltani-Sedeh1, Shiva Irani2, Reza Mirfakhraie3
1Department of Biology, Science and Research Branch, Islamic Azad University; Stem Cell Technology Research Center, Tehran, Iran.
Background:
MicroRNAs are small noncoding RNAs which modulate gene expression at different levels. It has been shown that downregulation of miR-34a occurs in varieties of cancers including colorectal cancer (CRC). In this study, we investigated the potential tumor inhibitory effects of miR-34a alone or in combination with paclitaxel in CRC cells.
Materials And Methods:
SW480 cells were transduced with lentiviral overexpressed miR-34a. First, using 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, the effect of miR-34a induction alone or in combination with paclitaxel on the cell viability and cell proliferation were estimated. Then, the expression level of target genes was measured using quantitative reverse transcription-polymerase chain reaction analysis. Eventually, the role of miR-34a and paclitaxel on cell cycle were determined with flow cytometry.
Results:
Gene expression analysis showed that miR-34a downregulates the expression of BCL2 and SIRT1 genes at mRNA level. Furthermore, miR-34a has a potential to reduce cell viability and cell cycle arrest at G1 phase. Combination of paclitaxel with overexpression of miR-34a significantly decreased cell viability compared to cell treated with miR-34a or paclitaxel alone. Interestingly, a combination of miR-34a and paclitaxel arrested cell cycle at two phases.
Conclusion:
Our results suggested that combination therapy of miR-34a and paclitaxel could be considered as the potential treatment of CRC.
Insights
Overexpressing miR-34a in colorectal cancer (CRC) cells inhibits tumor growth and enhances paclitaxel efficacy. This combination therapy shows potential for CRC treatment by reducing cell viability and arresting cell cycle.
Area of Science:
- Molecular Biology
- Oncology
- Biochemistry
Background:
- MicroRNAs regulate gene expression; miR-34a is downregulated in colorectal cancer (CRC).
- Investigating miR-34a's tumor-inhibitory role in CRC cells, alone and with paclitaxel.
Purpose of the Study:
- To evaluate the therapeutic potential of miR-34a in colorectal cancer.
- To assess the synergistic effect of miR-34a and paclitaxel combination therapy.
Main Methods:
- Overexpression of miR-34a in SW480 CRC cells using lentiviral vectors.
- Assessing cell viability, proliferation, target gene expression (BCL2, SIRT1), and cell cycle progression via MTT assay, qRT-PCR, and flow cytometry.
Main Results:
- miR-34a downregulated BCL2 and SIRT1 mRNA expression.
- miR-34a reduced cell viability and induced G1 phase arrest.
- Combination therapy significantly decreased cell viability and induced cell cycle arrest at multiple phases.
Conclusions:
- miR-34a demonstrates tumor-inhibitory effects in CRC.
- Combination of miR-34a and paclitaxel presents a promising therapeutic strategy for colorectal cancer.
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