Browsing by Author "Chung, W. Y."

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    Stearoyl-CoA desaturase 1 inhibitor supplemented with gemcitabine treatment reduces the viability and fatty acid content of pancreatic cancer cells in vitro
    (Wolters Kluwer, 2021-12) Hackney, A. B.; Chung, W. Y.; Isherwood, J.; Dennison, A. R.; Martin, N.
    Objective: Pancreatic cancer (PC) is an aggressive cancer with ineffective treatment. Inhibition of stearoyl-CoA desaturase 1 (SCD1) suppresses cancer proliferation and might act as a novel chemotherapy supplement, but this has not been investigated in PC. Here, the effects of SCD1 inhibitor CAY10566 supplemented with gemcitabine treatment (gemcitabine+CAY10566) on PC cell viability, apoptosis, phenotype, fatty acid content, platelet-derived growth factor release, and cell size were investigated. Methods: Human PC cell line (PANC-1) was treated with SCD1 inhibitor CAY10566 with or without gemcitabine. Cell viability was assayed using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide and apoptosis and phenotype were determined using flow cytometry. Fatty acid content and platelet-derived growth factor release were measured by enzyme-linked immunosorbent assay. Cell size was determined using scanning electron microscopy. Results: Half-maximal inhibitory concentration of gemcitabine or CAY10566 significantly reduced PANC-1 viability compared to gemcitabine alone (P<.0001). No significant differences in the phenotype of phosphatidylserine, tissue factor or basigin expression were detected at therapeutic doses (P>.05). Apoptosis was significantly increased following incubation with CAY10566 (P<.05). Fatty acid content of cells was significantly higher following gemcitabine treatment compared to CAY10566 alone or gemcitabine +CAY10566 (P<.05). Platelet-derived growth factor released by gemcitabine-treated cells was significantly increased compared to 142nM CAY10566 alone or gemcitabine+CAY10566 (P<.01). CAY10566 did not affect the size of isolated tumor cells but gemcitabine+CAY10566 significantly increased the size compared to the control (P<.05). Cell viability decreased significantly after the treatment with gemcitabine+CAY10566 compared with CAY10566 alone (P<.05) and gemcitabine alone (P<.01). However, when cycles of chemotherapy were mimicked and treatment was removed, the number of cell viability was significantly reduced (P<.05). Conclusion: This study suggests that CAY10566 may be a suitable supplement for gemcitabine chemotherapy for PC.
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    Stearoyl-CoA-Desaturase 1 inhibition induces apoptosis in Pancreatic Cancer cells
    (2018-06) Isherwood, J.; Runau, F.; Arshad, A.; Martin, N.; Hackney, A. B.; Chung, W. Y.; Dennison, A. R.
    Pancreatic cancer (PC) is an aggressive cancer with advanced disease at diagnosis and lack of effective therapy. PC tumours carry a mutated KRAS gene essential to their growth and spread, which plays a key role in deregulating cell metabolism. Stearoyl-CoA-Desaturase 1 (SCD1) is a highly regulated enzyme responsible for desaturating fatty acids, converting Stearic acid to Oleic acid. The expression of SCD1 is known to be upregulated in PC, indicating that it represents a metabolic bottleneck for cancer cell metabolism and might contribute to the growth and spread of PC. The aim of this study is to determine the effects of inhibiting SCD1 activity on a PC cell line (PANC1) in vitro. PANC1 cells were incubated for 48 hours with various concentrations of CAY10566, a selective SCD1 inhibitor. The cells were trypsinized, stained with Annexin V (AnV) and Propidium Iodide (PI) and analysed using flow cytometry. SCD1 inhibition (INHIB) significantly decreases viable PANC1 cells (AnV-, PI-) compared to a medium-only control (CON) (INHIB 48.76% (± 0.03), CON 60.71% (±0.01), P<0.05) and significantly increased apoptosis (AnV +, PI-) (INHIB 31.56% (±0.07), CON 25.32% (±0.02), P<0.05. Microscopically, the membrane of these cells appears less defined after treatment with the SCD inhibitor, indicating changes in composition may occur. Our results show that SCD1 inhibition significantly affects PC cell death and apoptosis. Further work is required to combine SCD1 inhibition with the standard PC chemotherapy Gemcitabine, and assess if there is a synergistic effect of the two drugs. This supplemented treatment could improve TMN (tumour metastasis node) and progression free survival, both clinical markers of patient outcome, in PC patients.
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    Stearoyl-CoA-Desaturase Inhibition Significantly Reduces the Growth of Pancreatic Cancer
    (2018-10) Martin, N.; Hackney, A. B.; Chung, W. Y.; Dennison, A. R.
    Aims: To determine the potency of Stearoyl-CoA-Desaturase 1 (SCD1) inhibition as a viable treatment option or chemotherapy supplementation for patients with Pancreatic Cancer and how it affects tumour-derived microparticle production. Method: PANC-1 cells were incubated with CAY10566 (0.125-2M), an inhibitor of SCD1 for 120hours. These were then counted, assayed for viability and the presence of Tissue Factor (CD142) and Basigin (CD147) was determined using flow cytometry. The migration of treated cells through a basement membrane was determined using Matrigel in a transwell invasion assay. The viability of cells treated with SCDi, Gemcitabine chemotherapy and both drugs in combination was assessed by MTT assay both immediately after incubation and after 48 hours recovery in complete medium to determine reversibility. Cells were also prepared and imaged using scanning electron microscopy (SEM) to observe any morphological changes induced by the treatments. Results: When stained for viability, no significant induction of apoptosis or necrosis by either drug was observed. When determined by MTT assay the IC50 values for SCDi and Gemcitabine were 142.4 and 13.05nm respectively. Cellular growth after recovery in complete medium (to mimic periods between treatment) was significantly lower after treatment with SCDi alone compared to SCDi in combination with Gemcitabine (38%2.19 SCDi v 153%9.45 SCDi+Gem)(P<0.05). SCDi treatment reduces the migration of CD147+ cells compared to both the vehicle control and Gemcitabine treatment (55.8% SCDi v 61% CON v 58% Gem). SEM imaging revealed that SCDi treated cells have a much flatter phenotype than vehicle control cells, and those treated with Gemcitabine and SCDi in combination are sparser, swollen in size (174m CON v 552m SCDi+Gem) and have a very flat phenotype. Conclusion: Both SCDi and Gemcitabine treatment affect the microscopic and flow cytometric phenotype of PANC1 cells. SCD1 inhibition is a promising treatment option for PC patients with a low IC50 value indicating high potency and a synergistic mechanism observed when used in combination with Gemcitabine. This indicates that treatment may be a suitable supplementation for standard therapy. Further work is required to elucidate the mechanism of growth inhibition induced by SCDi.