(a) Synthesis of compound CK21 as white solid after recrystallization in a mixed organic solvent. Compound structure was characterized by H-NMR, C-NMR, and HR-MS. (b) Thermal ellipsoid model …
Safety profile of CK21.
Acute maximum tolerated dose (MTD) studies, toxicity and toxicokinetic studies on rats and beagle dogs.
IC50 (µM) of triptolide (TP) or CK21 for different cancer cell lines in an in vitro cell viability assay.
(a) Scheme of in vivo efficacy studies. Human pancreatic cancer cell line, AsPc-1, was implanted into nude mice and CK21 treatment was initiated ~14 days later and administered daily for 4 weeks. (b)…
(a) Tumor growth with CK21 treatment at 3 mg/kg daily for 28 days. (b) Weight change of mice during Ck21 treatment.
(a) Subcutaneous AsPC-1 tumor in male mice after CK21 treatment at 1.5 mg/kg. (b) Male mice weight during CK21 treatment. (N=5 for each experimental group).
(a) Scheme of delayed therapy. Mice received CK21 at 3 mg/kg daily for 4 weeks, starting on day 42 post-tumor innoculation. (b) Tumor volume during delayed CK21 therapy. (c) Scheme of combination …
(a) In vitro culture of different organoids with escalating concentrations of CK21 for 72 hours. Gemcitabine was included as a positive control. (b) U049MAI tumor size in nude mice during CK21 …
Essential information on the pancreatic tumor organoids used in this study.
Details of organoids from Patient# 1, 2, 6, and 7 are provided in reference 39 (Romero-Calvo et al., 2019).
(a) CK21 (50 mM) reduced the expression of DDIT4 in AsPC-1, Panc-1, U049MAI, and U123M15-T after 24 hoursr of culture. (b) Baseline expression of DDIT4 in different tumor cells (without CK21 …
(a) knockdown of DDIT4 in Panc-1 did not alter response to CK21 (50 nM). (b) AsPC-1 overexpression of DDIT4 did not alter response to CK21 (50 nM).
CK21 given at 4 mg/kg/day for KC-6141 and 3 mg/kg/day for KPC-961 resulted in modest inhibition of tumor growth.
(a) Volcano plots of differentially expressed genes in (a) U049MAI and (b) U123M15-T after 12 hours CK21 treatment (50 nM). Significance cutoff was s P<0.05. Upregulation was colored as red, and …
Top pathways for (a) U049MAI and (b) U123M15-T.
(a) Heatmap of the relative expression of genes in the NF-κB pathway in U049MAI and U123M15-T after CK21 treatment. Genes are color coded where red means upregulated, and blue means downregulated. …
Full unedited gels.
Green represents downregulation and red represent upregulation by IPA analysis.
(a) Scheme of a subcutaneous model of mouse pancreatic tumor, KPC-960, with CK21 treatment. CK21 was provided at 3 mg/kg daily starting on day 5 or day 7. During secondary challenge, CK21 was …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Chemical compound, drug | CK21 | In house | NA | |
Chemical compound, drug | Gemcitabine | Actavis | 45963-619-59 | |
Cell line (Homo-sapiens) | AsPC-1 | ATCC | CRL-1682 | |
Cell line (Homo-sapiens) | Luciferase transfected AsPC-1 | Indiana University | N/A | Luciferase transfected |
Cell line (Homo-sapiens) | Panc-1 | ATCC | CRL-1469 | |
Cell line (Mus) | KC-6141 | University of Nebraska | N/A | |
Cell line (Mus) | KPC-960 | University of Nebraska | N/A | |
Cell line (Mus) | KPC-961 | University of Nebraska | N/A | |
Biological sample (Mus) | B6129SF1/J | Jackson Laboratory | 101043 | |
Biological sample (Mus) | C57BL/6 J | Jackson Laboratory | 000664 | |
Biological sample (Mus) | Athymic Nude-Foxn1nu | Envigo | ||
Commercial assay or kit | DMEM | ATCC | 30–2002 | |
Commercial assay or kit | RPMI | Quality Biological | 112-024-101 | |
Commercial assay or kit | Fetal bovine serum | Atlanta Biologicals | S115OH | |
Commercial assay or kit | Penicillin streptomycin | Gibco | 15140–122 | |
Commercial assay or kit | L-Glutamine | Gibco | 25030–081 | |
Commercial assay or kit | DMSO | Sigma | 276855 | |
Commercial assay or kit | Trypsin-EDTA | Stemcell | 07901 | |
Commercial assay or kit | TrypLE express | Gibco | 12605–010 | |
Commercial assay or kit | Sodium pyruvate | Gibco | 11360–070 | |
Commercial assay or kit | MEM nonessential amino acids | Cellgro | 25–025 CL | |
Commercial assay or kit | 2-Mercaptoethanol | Gibco | 21985–023 | |
Commercial assay or kit | IntestiCult organoid growth medium | Stemcell | 6005 | |
Commercial assay or kit | A83-01 | Sigma | SML0788 | |
Commercial assay or kit | FGF-10 | Sigma | SRP3262 | |
Commercial assay or kit | Gastrin I | Sigma | G9145 | |
Commercial assay or kit | N-acetylcysteine | Sigma | A9165 | |
Commercial assay or kit | Nicotinamide | Sigma | N0636 | |
Commercial assay or kit | B27 supplement | Gibco | 17504–044 | |
Commercial assay or kit | Primocine | Invivogen | ant-pm-1 | |
Commercial assay or kit | Y-27632 | Tocris | 1254 | |
Commercial assay or kit | Matrigel | Corning | 356231 | |
Commercial assay or kit | TrypLE | Gibco | 12605–010 | |
Commercial assay or kit | CellTiter 96 AQueous one solution | Promega | G3580 | |
Commercial assay or kit | Caspase-3/7 green detection | Thermo Fisher | C10427 | |
Commercial assay or kit | SYTOX dead cell stain | Thermo Fisher | C10427 | |
Commercial assay or kit | CFSE cell proliferation kit | Thermo Fisher | C34554 | |
Commercial assay or kit | ACK lysing buffer | Quality Biological | 118-156-101 | |
Commercial assay or kit | ROS-Glo H2O2 assay | Promega | G8820 | |
Commercial assay or kit | NuPAGE 10% Bis-Tris gel | Invitrogen | NP0301BOX | |
Commercial assay or kit | NuPAGE MES SDS running buffer | Novex | NP002 | |
Commercial assay or kit | NuPAGE MOPS SDS running buffer | Novex | NP001 | |
Commercial assay or kit | NuPAGE transfer buffer | Novex | NP0006-1 | |
Commercial assay or kit | NuPAGE LDS sample reducing agent | Invitrogen | NP0007 | |
Commercial assay or kit | NuPAGE sample buffer | Invitrogen | NP0009 | |
Commercial assay or kit | NuPAGE antioxidant | Invitrogen | NP0005 | |
Commercial assay or kit | TBS Tween-20 buffer | Thermo Scientific | 28360 | |
Commercial assay or kit | Invitrolon PVDF filter paper | Novex | LC2005 | |
Commercial assay or kit | PageRuler prestained protein ladder | Thermo Scientific | 26616 | |
Commercial assay or kit | Methanol | Fisher Scientific | A452-4 | |
Commercial assay or kit | Pierce protease&phosphatase inhibitor | Thermo Scientific | A32959 | |
Commercial assay or kit | Bovine serum albumin | Sigma | A7906 | |
Commercial assay or kit | SuperSignal west pico PLUS | Thermo Scientific | 34579 | |
Commercial assay or kit | Pierce bradford assay kit | Thermo Scientific | 23246 | |
Antibody | Anti-beta actin (Rabbit polyclonal) | Abcam | ab8227 | (1:2000) |
Antibody | Recombinant anti-REDD-1/DDIT4 (Rabbit monoclonal) | Abcam | ab191871 | (1:1000) |
Antibody | Anti-Caspase-3 (Rabbit polyclonal) | Abcam | ab13847 | (1:500) |
Antibody | Recombinant anti- BCL2 (Rabbit monoclonal) | Abcam | ab182858 | (1:2000) |
Antibody | Goat anti-rabbit IgG H&L (Goat polyclonal) | Abcam | ab205718 | (1:10000) |
Antibody | Phospho-NFkB p65, PE, eBioscience(Mouse monoclonal) | Invitrogen | 12986342 | (1:100) |
Commercial assay or kit | 4',6-Diamidino-2-Phenylindole, Dilactate | Biolegend | 422801 | (1:1000) |
Commercial assay or kit | PowerUp SYBR green master mix | Applied Biosystem | A25742 | |
Commercial assay or kit | High capacity cDNA reverse transcription | Applied Biosystem | 4368814 | |
Commercial assay or kit | D-Luciferin potassium salt | Perkin Elmer | 122799 | |
Commercial assay or kit | PBS | GenClone | 25–508 | |
Commercial assay or kit | Cell recovery solution | Corning | 354253 | |
Commercial assay or kit | RNeasy Plus Mini Kit | Qiangen | 74124 | |
Commercial assay or kit | DNase I recombinant | Roche | 04536282001 |