Vincristine

Manufactured by Selleck Chemicals

Sourced in United States

Vincristine is a laboratory chemical compound used in various research and analytical applications. It serves as a standard reference material and is commonly employed in chemical analysis and testing procedures.

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Vincristine (VCR Vincristine sulfate

49 protocols using vincristine

Evaluating Chemotherapeutic Efficacy in Cell Lines

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RD and K562 cells were transfected with either siRNAs or LNA-modified oligonucleotides as previously described. 24 h after transfection, an equal number of transfected live cells were plated into a 12-well plate. At the same time, the indicated concentrations of either Vincristine (Selleckchem), Taxol (Paclitaxel - Selleckchem) or Nocodazole (Sigma) were added to the medium.
SK-N-BE cells were transfected with either si-SCR or si-Circ as previously described.
24 h after transfection cells were plated in 96-well plates (3x10⁴ cells/well), and the following day treated with the indicated concentrations of either Taxol or Vincristine.
24 h after the addition of the drugs, viable RD and SK-N-BE cells were stained with crystal violet solution (0.5%) and then the absorbance was read at 600 nm, using a Glomax-Multi+ Detection System machine (Promega). Instead, dead/live K562 cells were counted with the addition of 0.4% Trypan Blue stain (ThermoFisher Scientific) at a CytoSmart cell counter (Corning). Survival was expressed as percentage of cells alive normalized to DMSO-treated cells (vehicle control), and untreated cells were used to control each condition.

Rossi F., Beltran M., Damizia M., Grelloni C., Colantoni A., Setti A., Di Timoteo G., Dattilo D., Centrón-Broco A., Nicoletti C., Fanciulli M., Lavia P, & Bozzoni I. (2022). Circular RNA ZNF609/CKAP5 mRNA interaction regulates microtubule dynamics and tumorigenicity. Molecular Cell, 82(1), 75-89.e9.

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Pharmacological inhibition assay

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All drugs were kept in −20°C dissolved in DMSO in stock concentrations and were thawed and diluted to their final work concentrations in relevant medium immediately before addition to cell cultures. Drugs were as follows: foretinib (50 mM stock and 500 nM experimental; Selleckchem), lestaurtinib (50 mM stock and 1 µM experimental; Tocris), JNK inhibitor VII, TAT-TI-JIP_153–163 (10 mM stock and 2 µM experimental; Calbiochem), 1NMPP1 (25 mM stock; Calbiochem), vincristine (100 µM stock and 40 nM experimental; Selleckchem), cisplatin (50 mM stock and 20 µM experimental; Selleckchem), ABT263 (1 mM stock and 10 µM experimental; Selleckchem), and Z-VAD-FMK (0.5 M stock and 100 µM experimental; Calbiochem).

Feinberg K., Kolaj A., Wu C., Grinshtein N., Krieger J.R., Moran M.F., Rubin L.L., Miller F.D, & Kaplan D.R. (2017). A neuroprotective agent that inactivates prodegenerative TrkA and preserves mitochondria. The Journal of Cell Biology, 216(11), 3655-3675.

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PTCL Cell Line Maintenance and Treatments

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Peripheral T-cell non-Hodgkin lymphoma (PTCL) murine cell lines TIB-48 and CRL-2396 (TK1) and human leukemia cell line Jurkat clone E6-1 (ATCC) and SUP-T1, DERL-2 (DSMZ) were maintained in RPMI medium (Mediatech, VA) supplemented with 10% fetal bovine serum, 2mM sodium pyruvate at 37°C in a humidified atmosphere containing 5% CO₂. Alisertib and MLN1117 were provided by Takeda pharmaceuticals (Cambridge, MA). PF-04691502, idelalisib, duvelisib and vincristine were purchased from SelleckChem (Houston, TX). Mouse Anti-PD-L1 (BE0101) antibodies were purchased from BioXCell (West Lebanon, NH).

Islam S., Vick E., Huber B., Morales C., Spier C., Cooke L., Weterings E, & Mahadevan D. (2017). Co-targeting aurora kinase with PD-L1 and PI3K abrogates immune checkpoint mediated proliferation in peripheral T-cell lymphoma: a novel therapeutic strategy. Oncotarget, 8(59), 100326-100338.

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Ferroptosis Inducers and Inhibitors

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Erastin2 (compound 35MEW28 in (Dixon et al., 2014 (link))) and ML162 were synthesized by Acme Bioscience (Palo Alto, CA, USA). Bortezomib (Cat# NC0587961), thapsigargin (Cat# T9033), N-Acetyl-L-Cysteine (Cat# A8199) and ferrostatin-1 (Cat# SML0583) were from Sigma-Aldrich. Buthionine sulfoximine (Cat# AC23552-0010), Q-VD-OPh (Cat# OPH00101M) and monochlorobimane (Cat# M-1381MP) were from Thermo Fisher. C11 BODIPY 581/591 (Cat# D3861) was from Molecular Probes (Eugene, OR) and dissolved in methanol. RSL3 (Cat # S8155), Doxorubicin (Cat# S1208), vincristine (Cat# S1241), BAY-11-7085 (Cat# S7352) and BAY-11-7821/7082 (Cat# S2913) were from Selleck Chemicals (Houston, TX). Nec-1s (Cat# 2263) was from BioVision (Milpitas, CA). Buthionine sulfoximine was dissolved directly into cell media. All other compounds were prepared as stock solutions in DMSO and stored prior to use at −20°C. Cyst(e)inase enzyme was purified as described and stored at −80°C until use (Cramer etal., 2017 (link)).

Cao J.Y., Poddar A., Magtanong L., Lumb J.H., Mileur T.R., Reid M.A., Dovey C.M., Wang J., Locasale J.W., Stone E., Cole S.P., Carette J.E, & Dixon S.J. (2019). A Genome-wide Haploid Genetic Screen Identifies Regulators of Glutathione Abundance and Ferroptosis Sensitivity. Cell reports, 26(6), 1544-1556.e8.

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Ferroptosis Inducers and Inhibitors

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Vincristine Cytotoxicity Assay in MB Cells

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Cells were seeded at a density of 10⁵ cells/well in 96-well plates for 16 h to facilitate adhesion, before the addition of vincristine (Sigma, Oakville, ON, Canada) at various concentrations. After 48 h, the fluorometric reagent Cell Titer Blue (Promega, Madison, WI, USA) was added and fluorescence (560_Ex/590_Em) measured on a spectrophotometer (PE Enspire, Waltham, MA, USA) after 4 h. In some experiments involving microglia co-culture conditioning, MB cells were treated with ruxolitinib, bazedoxifene, or SC144 (Selleckchem, Houston, TX, USA) during the wean-off three day period, prior to replating cells in 96-well plates for combination treatment of the same agent with vincristine. All assays were conducted as three replicates per treatment condition.

Sreenivasan L., Li L.V., Leclair P, & Lim C.J. (2022). Targeting the gp130/STAT3 Axis Attenuates Tumor Microenvironment Mediated Chemoresistance in Group 3 Medulloblastoma Cells. Cells, 11(3), 381.

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Organoid Chemotherapy and Immunotherapy Evaluation

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Organoids were subsequently treated after 7 days of culture. Chemotherapies included cisplatin (1, 10, 100 µM) (232120, Sigma Aldrich), doxorubicin (0.1, 1, 10 µM) (S1208, Selleckchem), vincristine (0.1, 1, 10 µM) (V8879 Sigma Aldrich), doxorubicin with cisplatin (0.1/1, 1/10, 10/100 µM), cisplatin with etoposide (S1125, Selleckchem) (1/0.1, 10/1, 100/10 µM) (Selleckchem), and doxorubicin with vincristine (0.1/0.1, 1/1, 10/10 µM). For immunotherapy treatment, 100 nM of pembrolizumab (A2002, Selleckchem), ipilimumab (A2001, Selleckchem) or nivolumab (A2005, Selleckchem) was used on both iPTOs and PTOs in parallel treatments. Media was removed from the wells and drug solutions mixed in culture media were added. Organoids remained in treated media solution for 72 h prior to endpoint viability assessment.

Forsythe S.D., Erali R.A., Laney P., Sivakumar H., Li W., Skardal A., Soker S, & Votanopoulos K.I. (2022). Application of immune enhanced organoids in modeling personalized Merkel cell carcinoma research. Scientific Reports, 12, 13865.

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Cell Viability Assay for Leukemia Drugs

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Cells (25,000 per well) were seeded in 100 ml of complete growth medium in 96-well plates and incubated with chemotherapeutic agents or vehicle. T-ALL cells were split every 48 hr and AML cells were split every 72 hr. Cell viability was assessed by counting viable cells based on trypan blue vital dye staining (Invitrogen), according to the manufacturer’s instructions. Chemotherapeutic drugs included: asparaginase (pegaspargase, Shire, Lexington, MA), dexamethasone (Sigma-Aldrich), vincristine (Selleckchem, Houston, TX), doxorubicin (Sigma-Aldrich), 6-mercaptopurine (Abcam, Cambridge, UK), CHIR99021 (Selleckchem), rapamycin (Selleckchem), RAD001 (Selleckchem), AZD2014 (Selleckchem), thapsigargin (Sigma-Aldrich) and Wnt3A (R&D systems, Minneapolis, MN). BRD0705 and BRD3731 were synthesized as described (Wagner et al., 2018 ). Caspase 3/7 activity was assessed using the Caspase Glo 3/7 Assay (Promega, Madison, WI) according to the manufacturer’s instructions.

Hinze L., Pfirrmann M., Karim S., Degar J., McGuckin C., Vinjamur D., Sacher J., Stevenson K.E., Neuberg D.S., Orellana E., Stanulla M., Gregory R.I., Bauer D.E., Wagner F.F., Stegmaier K, & Gutierrez A. (2019). Synthetic Lethality of Wnt Pathway Activation and Asparaginase in Drug-Resistant Acute Leukemias. Cancer cell, 35(4), 664-676.e7.

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Vascularized MPS Drug Effects Evaluation

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To demonstrate the application of the vascularized MPS for studies of drug effects, microfluidic devices and 2D cell cultures of endothelial and tumor cells were exposed to three test compounds. FDA-approved anti-cancer drugs Fluorouracil (5-FU, S1209; concentrations tested: 1, 10, 100, 300, 1000 μM), Vincristine (S1241; concentrations tested: 1, 10, 100, 300, 1000 nM), and Sorafenib (S7397; concentrations tested: 0.1, 1, 10, 30, 100 μM), were purchased from Selleckchem (Houston, TX). All test compounds were dissolved in 100% cell culture grade dimethyl sulfate (DMSO, Fisher Scientific) to 200 mM for 5-FU and Vincristine and 20 mM for Sorafenib (i.e., 200× of the top test concentration). Final concentration of DMSO vehicle was 0.5% v/v in all conditions. Drug or vehicle treatments were introduced to the microfluidic devices through cell culture medium channels and reservoirs on day 6 post-loading. The interstitial flow was maintained by transferring cell culture medium from the outlet reservoir to the inlet reservoir daily to re-establish the interstitial pressure. Cell culture medium with drugs and/or vehicle was replaced with fresh EGM-2 on day 8 and the devices were monitored for another 2 days until day 10.

Liu Y., Sakolish C., Chen Z., Phan D.T., Bender R.H., Hughes C.C, & Rusyn I. (2020). Human In Vitro Vascularized Micro-Organ and Micro-Tumor Models are Reproducible Organ-on-a-Chip Platforms for Studies of Anticancer Drugs. Toxicology, 445, 152601.

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Sensitizing Cancer Cells to PTX

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PTX, norcantharidin and cisplatin were purchased from Sigma-Aldrich. Vincristine and Cariporide (NHE1 inhibitor) were purchased from Selleck Chemicals. MG132 (proteasome inhibitor) was purchased from MedChemExpress Chemicals. Hepatocellular carcinoma HepG2 and Human bladder cancer UMUC-3 cell lines were routinely maintained in DMEM medium (Gibco BRL, Grand Island, NY, USA). The medium contains 10% fetal bovine serum, penicillin (100 U/ml) and streptomycin (100μg/ml). Cells were cultured at 37 °C in a balanced air humidified incubator with an atmosphere of 5% CO₂. HepG2 and UMUC-3 cells were continuously treated with 40nM and 80nM PTX respectively for 7 days. The residual cells were then partially released into medium with half of the initial concentration of PTX.

Jia M., Zheng D., Wang X., Zhang Y., Chen S., Cai X., Mo L., Hu Z., Li H., Zhou Z, & Li J. (2020). Cancer Cell enters reversible quiescence through Intracellular Acidification to resist Paclitaxel Cytotoxicity. International Journal of Medical Sciences, 17(11), 1652-1664.

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