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Px 478

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PX-478 is a laboratory reagent used for scientific research purposes. It functions as a hypoxia-inducible factor-1α (HIF-1α) inhibitor, which plays a role in cellular response to low oxygen conditions. The core function of PX-478 is to modulate HIF-1α activity in experimental settings.

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Lab products found in correlation

Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Cocl2, by Merck Group (3 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Dimethyl sulfoxide (dmso), by Biosharp (2 mentions) Anti hif 2α, by Cell Signaling Technology (2 mentions) Anti hif 1α, by Cell Signaling Technology (2 mentions) Ab51608, by Abcam (2 mentions) Ly294002, by Selleck Chemicals (2 mentions) Agi 6780, by Selleck Chemicals (2 mentions) Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions) H1299, by American Type Culture Collection (1 mentions) Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Normal rabbit igg, by Cell Signaling Technology (1 mentions) Mitosox red, by Thermo Fisher Scientific (1 mentions) Cm h2dcfda, by Thermo Fisher Scientific (1 mentions) Anti α tubulin, by Thermo Fisher Scientific (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Anti myc sc 40, by Santa Cruz Biotechnology (1 mentions) Anti histone h3, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

PX-478 2HCl (3 citations) PX-478 (S7612) (2 citations)

26 protocols using px 478

1

EPC Administration Mitigates Ischemia Reperfusion

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Animals were randomly divided into four groups designated as PBS, EPC, EPC plus PX-478-treated and PX-478-treated groups. 1 × 106 EPCs suspended in 100 μl PBS was administered at the speed of 20 μl per minute through the left jugular vein immediately after reperfusion. The same amount of PBS was administrated as a control. PX-478 (40 mg/kg, Selleck, Houston, TX, USA) was injected intraperitoneally immediately after EPC administration.
Geng J., Wang L., Qu M., Song Y., Lin X., Chen Y., Mamtilahun M., Chen S., Zhang Z., Wang Y, & Yang G.Y. (2017). Endothelial progenitor cells transplantation attenuated blood-brain barrier damage after ischemia in diabetic mice via HIF-1α. Stem Cell Research & Therapy, 8, 163.
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2

Cancer Cell Culture and Hypoxia Assay

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SMMC7721 (human liver carcinoma cells), HepG2 (human liver hepatocellular carcinoma cells) and SW480 (human colorectal carcinoma cells) were cultured in Dulbecco’s modified Eagle’s medium (DMEM), which was obtained from Gibco (Grand Island, NY, USA). MDA-MB-231 cells (human breast cancer cells) were cultured in minimum essential medium (MEM, Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS, Millipore, Billerica, MA, USA) and maintained at 37 °C in a humidified incubator with 5% CO2. All four types of cancer cells were obtained from the Shanghai Cell Bank of the Chinese Academy of Sciences (CAS). To establish hypoxic conditions, the culture medium was supplemented with 150 μM CoCl2 for 24 h. The compounds were dissolved in dimethyl sulfoxide (DMSO, BIOSHARP, Hefei, China). PX-478 (Selleck Chemicals, Houston, TX, USA), a selective HIF-1α inhibitor, was dissolved in ddH2O. The control group was treated with DMSO only under identical conditions.
Wu C.Z., Liu D.C., Guo X., Dai Y., Ma T., Li H.M, & Huo Q. (2018). Synthesis and Evaluation of Bakuchiol Derivatives as Potential Anticancer Agents. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(3), 515.
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3

Inhibition of HIF-1α in NSCLC Cells

Cited 1 time
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Human NSCLC cell lines A549, H1299, human umbilical vein endothelial cells (HUVECs) were purchased from the American Type Culture Collection (ATCC). Cells were cultured in the complete RPMI1640 medium containing 10% inactivated newborn bovine serum (FBS, HyClone, Logan, UT, USA) and 1% penicillin/streptomycin. They were cultured at 37° C in an incubator with saturated humidity and 5% CO2. The medium was changed every 2 to 3 days. Cells were trypsinized and sub-cultured with 0.25% trypsin during the logarithmic growth phase. To inhibit HIF-1α in A549 cells, SKPV-3 cells were treated with PX-478 (40 μM) (Selleck Chemicals, Houston, TX, USA) for 18 hours [55 (link)].
Zhang J., Han L., Yu J., Li H, & Li Q. (2021). miR-224 aggravates cancer-associated fibroblast-induced progression of non-small cell lung cancer by modulating a positive loop of the SIRT3/AMPK/mTOR/HIF-1α axis. Aging (Albany NY), 13(7), 10431-10449.
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4

Hypoxia-Induced Cancer Cell Response

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SMMC7721 (human liver carcinoma cells), HepG2 (human liver hepatocellular carcinoma cells) and SW480 (human colorectal carcinoma cells) were cultured in Dulbecco’s modified Eagle’s medium (DMEM), which was obtained from Gibco (Grand Island, NY, USA). MDA-MB-231 cells (human breast cancer cells) were cultured in minimum essential medium (MEM, also obtained from Gibco) supplemented with 10% foetal bovine serum (FBS, Millipore, USA) and maintained at 37 °C in a humidified incubator with 5% CO2. All four types of cancer cells were obtained from Shanghai Cell Bank, Chinese Academy of Sciences (CAS). To establish a hypoxic condition, the culture medium was supplemented with 150 µmol/l CoCl2 for 24 h. The compounds were dissolved in dimethyl sulfoxide (DMSO, BIOSHARP, Hefei, China). The PX-478 (Selleck Chemicals, Houston, USA), a selective HIF-1α inhibitor was dissolved in ddH2O. The control group was treated with DMSO only under identical conditions.
Liu D.C., Gao M.J., Huo Q., Ma T., Wang Y, & Wu C.Z. (2019). Design, synthesis, and apoptosis-promoting effect evaluation of novel pyrazole with benzo[d]thiazole derivatives containing aminoguanidine units. Journal of Enzyme Inhibition and Medicinal Chemistry, 34(1), 829-837.
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5

Reagents for Hypoxia Signaling Pathway

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Anti-SMYD3 (#ab199361) antibody was purchased from Abcam. Antibodies including anti-HIF1α (#36169), anti-VHL (#68547), anti-Histone H3 (#4499), anti-HIF2α (#7096), anti-ARNT (#5537), and normal rabbit IgG (#2729) were purchased from Cell Signaling Technology. Anti-ACTB (#AC026) antibody was purchased from ABclonal. Anti-HA (#901515) antibody was purchased from Covance. Anti-Myc (#SC-40) and anti-GAPDH (#SC-477242) antibodies were purchased from Santa Cruz Biotechnology. Anti-α-tubulin (#62204), Alexa Fluor 488 goat anti-rabbit IgG (#A11008), Alexa Fluor 594 goat anti-mouse IgG (#A11005), CM-H2DCFDA (#C6827), and MitoSOX Red (#M36008) were purchased from Thermo Fisher Scientific. CoCl2 (#C8661) and deferoxamine mesylate salt (#D9533) were purchased from Sigma. FG4592 (#S1007) and PX478 (#S7612) were purchased from Selleck. Cycloheximide (#HY-12320) was purchased from MCE.
Wang Z., Chen X., Fan S., Zhu C., Deng H., Tang J., Sun X., Jia S., Liao Q., Xiao W, & Liu X. (2022). Methyltransferase SMYD3 impairs hypoxia tolerance by augmenting hypoxia signaling independent of its enzymatic activity. The Journal of Biological Chemistry, 298(12), 102633.
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6

Hypoxia Detection and Cell Culture

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Calcium chloride (CaCl2), trans-4-(Aminomethyl) cyclohexanecarboxylicacid (AMCHA), dimethyl sulfoxide (DMSO), Ficoll-Paque density gradient medium, DAPI, and glutaraldehyde, were purchased from Sigma-Aldrich (Saint Louis, MO). Type I collagenase and Image-iTTM Green Hypoxia detection reagent and Triton X-100 were purchased from Thermo Fischer Scientific (Waltham, MA). Cell tracker DiO (excitation, 488 nm; emission, 525/50 nm) was purchased from Invitrogen (Carlsbad, CA). Drugs including PX-478 and Durvalumab were purchased from Selleck Chemicals (Houston, TX).
Bhattacharya S., Calar K., Evans C., Petrasko M, & de la Puente P. (2020). Bioengineering the Oxygen-Deprived Tumor Microenvironment Within a Three-Dimensional Platform for Studying Tumor-Immune Interactions. Frontiers in Bioengineering and Biotechnology, 8, 1040.
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7

Cell Line Culturing and Compound Treatments

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The human malignant T-cell lines, Hut78, HH and Jurkat, and human malignant B-cell lines Daudi, Toledo, were obtained from the American Tissue Culture Collection (ATCC, Manassas, VA). Human CTCL line MyLa was generous gift from Dr. Reinhard Dummer, Dr. Emmanuel Contassot, and Dr. Lars French (University of Zurich). HH, Jurkat, Myla, Daudi and Toledo were propagated in RPMI-1640 media (ATCC) and Hut78 was propagated in Iscove's Modified Dulbecco's Medium (IMDM; ATCC). The following compounds were added to the media: 10% heat-inactivated fetal bovine serum, 100 U/mL penicillin, 0.1 mg/mL streptomycin, and 0.1 mg/mL Amphotericin B (Gibco, Gaithersburg, MD). Cells were kept in a humidified incubator at 37°C with a 5% CO2 atmosphere. Normal CD4+ T-cells (NTCs) from healthy donors were obtained from Astarte Biologics, Bothell, WA and maintained in X-VIVO15 (Lonza) media supplemented with human CD3/CD28/CD2 T cell activator (Immunocult, Stemcell Technologies, Cambridge, MA) and 50 IU/mL IL2 (BioVision, Milpitas, CA). Where indicated, cells were incubated with phenformin (10 μM), metformin (5 mM), N-acetylcysteine (4 mM), mitoTEMPO (30 μM), rotenone (1 μM), oligomycin (1 μg/mL), CoCl2 (100 μM) all from Sigma-Aldrich, St. Louis; MO, PX-478 (30 μM), MG-132 (25 μM), and IOX2 (50 μM), are from Selleckchem, Houston, TX.
Khan H., Anshu A., Prasad A., Roy S., Jeffery J., Kittipongdaja W., Yang D.T, & Schieke S.M. (2019). Metabolic Rewiring in Response to Biguanides is Mediated by mROS/HIF-1a in Malignant Lymphocytes. Cell reports, 29(10), 3009-3018.e4.
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8

Investigating Cellular Metabolic Pathways

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Rosiglitazone (5-[[4-[2-(Methyl-2-pyridinylamino)ethoxy]phenyl]methyl]-2,4-thiazolidinedione) and AICAR (5-Aminoimidazole-4-carboxamide 1-β-D-ribofuranoside) were purchased from Sigma and Tocris, respectively. GSK0660(3-(((2-Methoxy-4-(phenylamino)phenyl)amino]sulfonyl)-2-thiophenecarboxylic acid methyl ester) was purchased from abcam. PX-478 was purchased from Selleckchem. Oligonucleotides (Table S3) were purchased from Sigma and Qiagen.
Boutoual R., Meseguer S., Villarroya M., Martín-Hernández E., Errami M., Martín M.A., Casado M, & Armengod M.E. (2018). Defects in the mitochondrial-tRNA modification enzymes MTO1 and GTPBP3 promote different metabolic reprogramming through a HIF-PPARγ-UCP2-AMPK axis. Scientific Reports, 8, 1163.
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9

Immunohistochemistry and Western Blot of PLOD2 and HIF-1α/FAK

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Antibody to PLOD2 for immunohistochemistry and Western blot was purchased from Proteintech (Wuhan, China). HIF-1α (PX-478) and FAK inhibitors (TAE226; Selleckchem; Shanghai, China) were resolubilized in DMSO. Matrigel was purchased from BD Biosciences (San Diego, CA, USA). The primary human antibodies used in this study were the following: rabbit anti-HIF-1α, anti-HIF-2α, anti-FAK-p397, anti-FAK, Paxillin (Abcam; Cambridge, MA, USA); mouse anti-β-actin (Beyotime, China).
Xu Y., Zhang L., Wei Y., Zhang X., Xu R., Han M., Huang B., Chen A., Li W., Zhang Q., Li G., Wang J., Zhao P, & Li X. (2017). Procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 promotes hypoxia-induced glioma migration and invasion. Oncotarget, 8(14), 23401-23413.
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10

Prostate Organoids Treated with Enzalutamide

Cited 2 times
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Prostate organoid cultures were established from Pten(i)pe−/− and Pten/Hif1a(i)pe−/− mice at 3 months AGI and cultured as described (Karthaus et al, 2014 (link); Drost et al, 2016 (link); Abu El Maaty et al, 2022 (link)) and treated for 24 h with enzalutamide (MDV3100; Selleckchem; catalog No.S1250) at 25 and 50 μM, or vehicle (DMSO). C4‐2B (CRL‐3315), DU‐145 (HTB‐81), and PC‐3 (CRL‐1435) cells were obtained from ATCC and cultured in DMEM (4.5 g/l glucose, 10% FCS, 1% penicillin/streptomycin) at 37°C and 5% CO2. Cell lines used were mycoplasma‐free but not recently authenticated. HIF1A silencing in PC‐3 cells was performed as described (Abu El Maaty et al, 2022 (link)).
C4‐2B cells were treated for 24 h with PX‐478 (Selleck Chemicals; catalog No.S7612) at 50 μM and/or with enzalutamide (MDV3100; Selleckchem; catalog No.S1250) at 10, 25 and 50 μM, or vehicle (DMSO). DU‐145 and PC‐3 cells were treated for 24 h with PX‐478 at 50 μM.
Terzic J., Abu el Maaty M.A., Lutzing R., Vincent A., El Bizri R., Jung M., Keime C, & Metzger D. (2023). Hypoxia‐inducible factor 1A inhibition overcomes castration resistance of prostate tumors. EMBO Molecular Medicine, 15(6), e17209.
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