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Published on: November 7, 2019
Untargeted Metabolomics Analysis Reveals Potential Metabolic Targets in Gemcitabine-Treated Pancreatic Cancer Cells
Arjun Prasad Tiwari1, Blake R Rushing2,3, Larissa Silva1
1Department of Biological Sciences, University of North Carolina at Charlotte, Charlotte, NC 28223, USA.
Background/Objectives:
Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy characterized by limited treatment options and poor prognosis. Gemcitabine is a commonly used chemotherapy; however, gemcitabine resistance in PDAC poses a critical barrier to effective treatment, as the underlying mechanisms are not yet fully understood.
Methods:
This study employs an exploratory untargeted metabolomics approach to investigate metabolic differences in PDAC cells in the presence and absence of gemcitabine treatment. HPAF-II, MIA PaCa-2, and BxPC-3 cell lines were used as models for gemcitabine-resistant, moderately responsive, and permissive PDAC cells, respectively.
Results:
MTT assay results revealed that BxPC-3 cells are highly sensitive to gemcitabine treatment, HPAF-II cells are the most resistant, and MIA PaCa-2 cells exhibit moderate sensitivity. Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) of the metabolomics data demonstrated clear differentiation of gemcitabine-treated and untreated (control) cells. When comparing the treated vs. control conditions, 170 metabolites matched to an in-house library of standards were significant (p < 0.05 or fold change ≥ 2 or VIP ≥ 1) differentiators in HPAF-II cells, whereas MIA PaCa-2 and BxPC-3 cells had 178 and 218 differentiating metabolites, respectively. HPAF-II cells treated with gemcitabine had significantly higher levels of N-acetylneuraminic acid and 7-dehydrocholesterol compared with the control group. In contrast, these metabolites were significantly lower or non-significant in BxPC-3 treated cells. Pathway analysis revealed that the steroid biosynthesis pathway was significantly perturbed in HPAF-II cells, whereas amino sugar and nucleotide sugar metabolism was predominantly altered in BxPC-3 cells.
Conclusions:
Overall, this exploratory study reveals metabolic differences between treated and untreated cells to derive targeted therapeutic strategies that could be used in the future to improve treatment outcomes for PDAC patients.
Insights
This study explored metabolic differences in pancreatic ductal adenocarcinoma (PDAC) cells treated with gemcitabine. Resistant PDAC cells showed altered steroid biosynthesis, suggesting new therapeutic targets.
Area of Science:
- Metabolomics
- Cancer Biology
- Biochemistry
Background:
- Pancreatic ductal adenocarcinoma (PDAC) is aggressive with poor prognosis.
- Gemcitabine resistance in PDAC limits treatment efficacy.
- Mechanisms of gemcitabine resistance are not fully understood.
Purpose of the Study:
- Investigate metabolic differences in PDAC cells with and without gemcitabine treatment.
- Identify metabolic alterations associated with gemcitabine resistance.
- Explore potential therapeutic strategies for PDAC.
Main Methods:
- Untargeted metabolomics approach.
- Utilized three PDAC cell lines (HPAF-II, MIA PaCa-2, BxPC-3) representing varying gemcitabine sensitivity.
- Analyzed metabolic profiles using Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA).
Main Results:
- Significant metabolic differences observed between gemcitabine-treated and untreated PDAC cells.
- Resistant HPAF-II cells showed elevated N-acetylneuraminic acid and 7-dehydrocholesterol.
- Steroid biosynthesis pathway was perturbed in resistant cells, while amino sugar metabolism was altered in sensitive cells.
Conclusions:
- Exploratory study reveals distinct metabolic profiles in gemcitabine-treated PDAC cells.
- Identified specific metabolic alterations linked to gemcitabine resistance.
- Findings may inform future targeted therapeutic strategies for PDAC.
