sihk2  (Santa Cruz Biotechnology)

Journal: Nutrients

Article Title: Curcumin and Its Potential to Target the Glycolytic Behavior of Lactate-Acclimated Prostate Carcinoma Cells with Docetaxel

doi: 10.3390/nu16244338

Figure Lengend Snippet: Mitochondrial localization of HK2 and effect of docetaxel treatment on PC-3AcT and DU145AcT cells. Cells were cultured in DMEM containing 3.8 μM lactic acid with or without docetaxel (40 nM) for 48 h. ( A ) Western blot analysis of HK2 in mitochondrial and cytosolic fractions. ( B ) Western blot analysis of complexes I–V in the mitochondrial electron transport chain. ( C ) Measurement of mitochondrial membrane potential after staining cells with rhodamine123. ( D ) Changes in intracellular ATP concentration. ( E ) Percent cell viability for cells treated with or without 40 nM docetaxel. ( F ) Annexin V-PE binding assay for cells treated with or without 40 nM docetaxel. ( G ) Measurements of mitochondrial membrane potential for cells treated with or without 40 nM docetaxel. Data are expressed as the mean ± standard deviation of three independent experiments. Statistical significance comparing respective PC-3 or DU145 cells was considered at * p < 0.05 using one-way ANOVA and Tukey’s post hoc correction. HK2, hexokinase 2; VDAC, voltage-dependent anion channel; NDUFB8, NADH-ubiquinone oxidoreductase subunit B8 (complex I); SDHB, succinate dehydrogenase complex iron sulfur subunit B (complex II); UQCRC2, ubiquinone-cytochrome C reductase core protein 2 (complex III); COX II, mitochondrial cytochrome C oxidase subunit II (complex IV); ATP5A, ATP synthase F1 subunit alpha (complex V); DTX, docetaxel.

Article Snippet: RNA interference of HK2 was performed using a HK2-targeting small interfering RNA (siHK2) duplex from Invitrogen (Oligo ID, HSS179239).

Techniques: Cell Culture, Western Blot, Membrane, Staining, Concentration Assay, Binding Assay, Standard Deviation

Journal: Nutrients

Article Title: Curcumin and Its Potential to Target the Glycolytic Behavior of Lactate-Acclimated Prostate Carcinoma Cells with Docetaxel

doi: 10.3390/nu16244338

Figure Lengend Snippet: Effect of HK2 knockdown alone or in combination with curcumin on glucose metabolism in PC-3AcT and Du145AcT cells. Cells were transfected with 10 nM HK2-targeting siRNA (siHK2) or stealth RNAi control (siC) for 24 h. They were then treated with or without curcumin (40 μM) in DMEM containing 3.8 μM lactic acid for 48 h. ( A ) Percent cell viability. ( B ) Western blot analysis of key regulatory enzymes in glycolysis. ( C ) Activities of hexokinase and pyruvate dehydrogenase. ( D ) Changes in glucose concentration in culture medium. ( E ) Western blot analysis of HK2 in mitochondrial and cytosolic fractions. The bar graph represents densitometric analysis of Western blot images normalized to β-actin. Data are expressed as the mean ± standard deviation of three independent experiments. Statistical significance comparing the respective siC group was considered at * p < 0.05 using one-way ANOVA and Tukey’s post hoc correction. CCM, curcumin; HK, hexokinase; PFKP, phosphofructokinase platelet; PDH, pyruvate dehydrogenase.

Article Snippet: RNA interference of HK2 was performed using a HK2-targeting small interfering RNA (siHK2) duplex from Invitrogen (Oligo ID, HSS179239).

Techniques: Knockdown, Transfection, Control, Western Blot, Concentration Assay, Standard Deviation

Journal: Nutrients

Article Title: Curcumin and Its Potential to Target the Glycolytic Behavior of Lactate-Acclimated Prostate Carcinoma Cells with Docetaxel

doi: 10.3390/nu16244338

Figure Lengend Snippet: Effects of HK2 knockdown alone or in combination with curcumin on mitochondrial function and programmed cell death in PC-3AcT and Du145AcT cells. Cells were transfected with 10 nM HK2-targeting siRNA (siHK2) or stealth RNAi control (siC) for 24 h. They were then treated with or without curcumin (40 μM) in DMEM containing 3.8 μM lactic acid for 48 h. ( A ) Western blot analysis of complexes I–V in mitochondrial electron transport chain. ( B ) Measurements of mitochondrial membrane potential after staining cells with rhodamine123. ( C ) Changes in intracellular ATP concentration. ( D ) Cell cycle analysis. ( E ) Annexin V-PE binding assay. ( F ) Western blot analysis of apoptosis- and necroptosis-related proteins. Data are expressed as the mean ± standard deviation of three independent experiments. Statistical significance comparing the respective siC group was considered at * p < 0.05 using one-way ANOVA and Tukey’s post hoc correction. CCM, curcumin; NDUFB8, NADH-ubiquinone oxidoreductase subunit B8 (complex I); SDHB, succinate dehydrogenase complex iron sulfur subunit B (complex II); UQCRC2, ubiquinone-cytochrome C reductase core protein 2 (complex III); COX II, mitochondrial cytochrome C oxidase subunit II (complex IV); ATP5A, ATP synthase F1 subunit alpha (complex V).

Article Snippet: RNA interference of HK2 was performed using a HK2-targeting small interfering RNA (siHK2) duplex from Invitrogen (Oligo ID, HSS179239).

Techniques: Knockdown, Transfection, Control, Western Blot, Membrane, Staining, Concentration Assay, Cell Cycle Assay, Binding Assay, Standard Deviation