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Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
Genome-wide profiling of micro-RNA expression in gefitinib-resistant human lung adenocarcinoma using microarray for
Yong Hu1, Xiaobing Qin1,2, Dali Yan1
11 Department of Clinical Cancer Research Center, Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, Nanjing Medical University Affiliated Cancer Hospital, Nanjing, China.
Abstract:
To understand the mechanism involved in gefitinib resistance, we established gefitinib-resistant human HCC827/GR-8-1 cell line from the parental HCC827 cell line. We compared the micro-RNA expression profiles of the HCC827 cells HCC827/GR-8-1 using Agilent micro-RNA microarrays. The micro-RNAs, such as the miR-149-5p, were up- or downregulated and associated with acquired gefitinib resistance. Quantitative real-time polymerase chain reaction was then performed to verify the expression patterns of different micro-RNAs. The result showed that miR-149-5p was upregulated in the HCC827/GR-8-1 cell line. To investigate the biological function of miR-149-5p in non-small cell lung cancer cells acquired gefitinib resistance, we examined cell proliferation using a cell counting kit-8 assay. Cell viability was evaluated after the miR-149-5p mimics, inhibitors, and negative control were separately transfected into the non-small cell lung cancer cells. The results showed that the non-small cell lung cancer cells transfected with miR-149-5p mimics exhibited reduced cell motility. The drug-sensitivity assay results revealed that the overexpression of miR-149-5p effectively evaluates the half maximal inhibitory concentration values of the cell in response to gefitinib, and the downregulation of miR-149-5p can attenuate the half maximal inhibitory concentration values of the cell lines in response to gefitinib. Furthermore, the levels of miR-149-5p in the HCC827 and HCC827/GR-8-1 cells were inversely correlated with caspase-3 expression. In conclusion, this study revealed that miR-149-5p is upregulated in the HCC827/GR-8-1 cells and involved in the acquired gefitinib resistance.
Insights
This study found that miR-149-5p is upregulated in gefitinib-resistant lung cancer cells and contributes to acquired resistance. Upregulating miR-149-5p reduces cell motility and increases gefitinib sensitivity.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Gefitinib resistance is a significant challenge in non-small cell lung cancer (NSCLC) treatment.
- Understanding the molecular mechanisms underlying acquired gefitinib resistance is crucial for developing effective therapeutic strategies.
Purpose of the Study:
- To investigate the role of micro-RNAs (miRNAs) in acquired gefitinib resistance in NSCLC.
- To identify specific miRNAs associated with gefitinib resistance and elucidate their functional significance.
Main Methods:
- Established a gefitinib-resistant human HCC827/GR-8-1 cell line from parental HCC827 cells.
- Compared miRNA expression profiles using Agilent miRNA microarrays.
- Verified miRNA expression patterns via quantitative real-time polymerase chain reaction (qRT-PCR).
- Assessed cell proliferation and viability using cell counting kit-8 assay after miRNA mimic and inhibitor transfections.
- Evaluated drug sensitivity through half maximal inhibitory concentration (IC50) assays.
- Correlated miRNA levels with caspase-3 expression.
Main Results:
- Identified miR-149-5p as significantly upregulated in the gefitinib-resistant HCC827/GR-8-1 cell line compared to parental cells.
- Overexpression of miR-149-5p in NSCLC cells led to reduced cell motility.
- miR-149-5p mimics increased gefitinib sensitivity (lowered IC50 values), while inhibitors decreased sensitivity (raised IC50 values).
- Levels of miR-149-5p were inversely correlated with caspase-3 expression.
Conclusions:
- miR-149-5p is upregulated in acquired gefitinib-resistant NSCLC cells.
- miR-149-5p plays a critical role in mediating acquired gefitinib resistance by affecting cell motility and gefitinib sensitivity.
- miR-149-5p represents a potential therapeutic target for overcoming gefitinib resistance in NSCLC.
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