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Published on: February 21, 2014
Integrative gene expression profiling reveals that dysregulated triple microRNAs confer paclitaxel resistance in
Yuanming Cai1, Ruxue Jia2,3, Haozhe Xiong4
1Undergraduate Grade 2015, The 5th Clinical Medical School of Xinjiang Medical University, Urumchi, People's Republic of China.
Background:
Paclitaxel has shown significant anti-tumor activity against non-small cell lung cancer (NSCLC); however, resistance to paclitaxel frequently occurs and represents a significant clinical problem and its underlying molecular mechanism remains elusive.
Methods:
Long-term treatment of culture cell with paclitaxel was carried out to mimic the development of acquired drug resistance in NSCLC. Cell proliferation and clonogenic assay and apoptosis evaluation were carried out to determine the efficacy of paclitaxel on NSCLC cells. Western blot analyses were performed to determine the expression and activation of proteins. Apoptosis enzyme-linked immunosorbent assay was used to quantify cytoplasmic histone-associated DNA fragments. Microarray analyses were applied to explore both mRNA and miRNA expression profiles in NSCLC cells followed by integrative analysis. qRT-PCR was carried out to verify the differentially expressed mRNAs and miRNAs.
Results:
The expression of 652 genes was shown to be changed at least 2-fold in paclitaxel-resistant NSCLC (H460_TaxR) cells with 511 upregulated and 141 downregulated as compared with that in parental H460 cells. The differentially expressed genes were functionally enriched in regulating the cell proliferation, cell death, and response to endogenous stimulus, and clustered in pathways such as cancer and signaling by the G protein-coupled receptor (GPCR). Moreover, 43 miRNAs were shown to be differentially expressed in H460_TaxR cells with 15 upregulated and 28 downregulated as compared with parental H460 cells. A total of 289 pairs of miRNA-potential target gene were revealed in H460_TaxR cells by bioinformatics analysis. Furthermore, integrative analysis of miRNAs and gene expression profiles revealed that dysregulated miR-362-3p, miR-766-3p, and miR-6507-3p might confer paclitaxel resistance in NSCLC via targeting MAPT simultaneously.
Conclusion:
Our findings suggested that specific manipulation of MAPT-targeting miRNAs may be a novel strategy to overcome paclitaxel resistance in patients with NSCLC especially large-cell lung carcinoma.
Insights
Paclitaxel resistance in non-small cell lung cancer (NSCLC) was investigated. Specific microRNAs targeting MAPT were identified as potential therapeutic targets to overcome this resistance.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Paclitaxel exhibits anti-tumor activity in non-small cell lung cancer (NSCLC).
- Acquired paclitaxel resistance in NSCLC is a significant clinical challenge with an elusive molecular mechanism.
Purpose of the Study:
- To investigate the molecular mechanisms underlying paclitaxel resistance in NSCLC.
- To identify potential therapeutic targets for overcoming paclitaxel resistance.
Main Methods:
- Developed paclitaxel-resistant NSCLC cell lines (H460_TaxR) through long-term drug exposure.
- Utilized cell proliferation, clonogenic, and apoptosis assays to assess drug efficacy.
- Performed Western blot, apoptosis ELISA, and microarray analyses to examine protein and gene expression profiles (mRNA and miRNA).
- Conducted integrative bioinformatics analysis to identify miRNA-mRNA interactions.
Main Results:
- Paclitaxel-resistant NSCLC cells exhibited significant alterations in gene expression, with 511 upregulated and 141 downregulated genes.
- Differentially expressed genes were enriched in pathways related to cell proliferation, cell death, and GPCR signaling.
- 43 miRNAs were differentially expressed in resistant cells, with 15 upregulated and 28 downregulated.
- Integrative analysis identified dysregulated miR-362-3p, miR-766-3p, and miR-6507-3p as potential mediators of paclitaxel resistance by targeting MAPT.
Conclusions:
- Specific microRNAs (miR-362-3p, miR-766-3p, miR-6507-3p) targeting MAPT are implicated in paclitaxel resistance in NSCLC.
- Targeting these MAPT-targeting miRNAs presents a novel therapeutic strategy to overcome paclitaxel resistance, particularly in large-cell lung carcinoma.
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