Abstract
Background
The main therapies for cancer often results in many side effects and drug resistance. Gamma linolenic acid (GLA) is a kind of natural reagent with negligible cytotoxicity.
Objective
This work aims at detecting whether GLA possesses anti-cancer activity in NSCLC cells and elucidating the potential molecular mechanism.
Methods
Cytotoxicity of GLA was evaluated by MTT assay and soft agar colony formation method. Immunoblotting analysis examined the effect of GLA on protein expressions of cell proliferation markers (e.g., PCNA, Ki-67 and MCM2), pro-survival protein bcl-2, apoptosis-associated proteins (e.g., bax and cleaved caspase 3), HIF1α and VEGF. Wound healing assay and transwell invasion assay were performed to test the effect of GLA on hypoxia-induced cell migration and invasion. Cell transfection was used to overexpress HIF1α followed by the treatment of GLA to test the effect of HIF1α overexpression on the tumoricidal activity of GLA in NSCLC cell lines.
Results
MTT and soft agar colony formation tests showed that GLA dose-dependently suppressed cell proliferation in both Calu-1 and SK-MES-1 cell lines. Immunoblotting analysis demonstrated that GLA suppressed protein expressions of PCNA, Ki-67, MCM2 and bcl-2, while GLA induced bax and cleaved caspase 3 expressions. Wound healing assay and transwell invasion assay revealed that GLA was very effective on the inhibition of NSCLC cell migration and invasion. Immunoblotting analysis and cell transfection method indicated that GLA inhibited hypoxia-induced cell proliferation and invasion by suppressing HIF1α-VEGF pathway.
Conclusion
GLA suppresses hypoxia-induced proliferation and invasion of NSCLC cells by inhibition of HIF1α pathway in vitro.
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Data availability
All data generated or analyzed during this study are included in this published article.
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YW conceived and designed the experiments, JS analyzed and interpreted the results of the experiments, LYG performed the experiments.
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Yan Wang, Jian Shi and Liya Gong declare that they have no conflict of interest.
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Wang, Y., Shi, J. & Gong, L. Gamma linolenic acid suppresses hypoxia-induced proliferation and invasion of non-small cell lung cancer cells by inhibition of HIF1α. Genes Genom 42, 927–935 (2020). https://doi.org/10.1007/s13258-020-00961-5
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DOI: https://doi.org/10.1007/s13258-020-00961-5