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Microarray-based Identification of Individual HERV Loci Expression: Application to Biomarker Discovery in Prostate Cancer
Published on: November 2, 2013
Microarray-based identification of differentially expressed genes associated with andrographolide derivatives-induced
Shun Ying Quah1, Charng Choon Wong1, Hui Chyn Wong1
1Pharmacotherapeutics Unit, Department of Medicine, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Abstract:
Chemoresistance poses a major hurdle to cancer treatments. Andrographolide-derived SRJ09 and SRJ23 were reported to exhibit potent, selective inhibitory activities against colon and prostate cancer cells, respectively. In this study, previously developed resistant colon (HCT-116rst09) and prostate (PC-3rst23) cancer cell lines were used to elucidate the molecular mechanisms contributing to chemoresistance. Cytotoxic effects of SRJ09 and SRJ23 on both parental and resistant cells were investigated. Cell cycle distributions in HCT-116rst09 cells following SRJ09 treatment were analysed using flow cytometry. Whole-genome microarray analysis was performed on both parental and resistant cells to obtain differential gene expression profiles. Microarray data were subjected to protein-protein interaction network, functional enrichment, and pathway analyses. Reverse transcription-polymerase chain reaction (RT-PCR) was used to validate the changes in expression levels of selected genes. Besides morphological changes, HCT-116rst09 cells showed 7.0-fold resistance to SRJ09 while PC-3rst23 cells displayed a 5.5-fold resistance to SRJ23, as compared with their respective parental cells. G0/G1-phase cell cycle arrest was observed in HCT-116rst09 cells upon SRJ09 treatment. Collectively, 77 and 21 genes were found differentially modulated in HCT-116rst09 and PC-3rst23 cells, respectively. Subsequent bioinformatics analysis revealed several genes associated with FGFR4 and PI3K pathways, and cancer stemness, were chemoresistance mediators in HCT-116rst09 cells. RT-PCR confirmed the HMOX1 upregulation and ATG12 downregulation protected the PC-3rst23 cells from SRJ23 cytotoxicity. In conclusion, acquired chemoresistance to SRJ09 and SRJ23 in colon and prostate cancer cells, respectively, could be attributed to the alterations in the expression of genes such as those related to PI3K and autophagy pathways.
Insights
Chemoresistance in colon and prostate cancer cells involves altered gene expression. Specific molecular pathways like PI3K and autophagy contribute to resistance against SRJ09 and SRJ23, impacting treatment efficacy.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Chemoresistance is a significant challenge in cancer therapy.
- Andrographolide derivatives SRJ09 and SRJ23 show promise against colon and prostate cancers.
- Understanding resistance mechanisms is crucial for improving treatment outcomes.
Purpose of the Study:
- To investigate the molecular mechanisms underlying acquired chemoresistance to SRJ09 in colon cancer cells (HCT-116) and SRJ23 in prostate cancer cells (PC-3).
- To identify key genes and pathways involved in mediating this resistance.
Main Methods:
- Development of resistant cell lines (HCT-116rst09 and PC-3rst23).
- Assessment of cytotoxicity and cell cycle effects.
- Whole-genome microarray analysis for differential gene expression.
- Bioinformatics analyses including pathway and protein-protein interaction network analysis.
- Validation using Reverse Transcription-Polymerase Chain Reaction (RT-PCR).
Main Results:
- Resistant cells exhibited significant resistance (7.0-fold for HCT-116rst09 to SRJ09, 5.5-fold for PC-3rst23 to SRJ23).
- SRJ09 induced G0/G1 cell cycle arrest in HCT-116rst09 cells.
- Differential gene expression identified 77 genes in HCT-116rst09 and 21 genes in PC-3rst23.
- Key pathways implicated include FGFR4, PI3K, cancer stemness, and autophagy.
- HMOX1 upregulation and ATG12 downregulation were confirmed in SRJ23-resistant cells.
Conclusions:
- Acquired chemoresistance to SRJ09 and SRJ23 in colon and prostate cancer cells is linked to significant alterations in gene expression.
- The PI3K and autophagy pathways play critical roles in mediating chemoresistance.
- Identifying these molecular mediators provides potential targets for overcoming drug resistance in cancer treatment.
