Bleomycin Sulfate

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Biological Activity

Description	Bleomycin Sulfate is a glycopeptide antibiotic and an anticancer agent for squamous cell carcinomas (SCC) with IC50 of 4 nM in UT-SCC-19A cells.
Targets	UT-SCC-19A cells
IC50	4 nM [1]
In vitro	UT-SCC-12A and UT-SCC-12B are both more resistant to Bleomycin sulfate with IC50 of 14.2 nM and 13 nM, respectively. [1] Alveolar macrophages incubated with 0.01 μg/mL to 1μg /mL Bleomycin sulfate for 18 hours secretes significantly more fibroblast growth factor than macrophages incubated without Bleomycin sulfate. Macrophages stimulated with Bleomycin sulfate continues to produce significant amounts of fibroblast growth factor even after Bleomycin sulfate is removed and replaced with fresh (Bleomycin sulfate-free) media. Fibroblast growth factor secretion by Bleomycin sulfate-stimulated alveolar macrophages is inhibited by cycloheximide, and the 5-lipoxygenase inhibitors NDGA (nordihydroguairetic acid) and BW755c, indicating not only a requirement for protein synthesis but also for metabolites of the 5-lipoxygenase pathway of arachidonic acid metabolism for full expression of activity. [2] Bleomycin sulfate (400 µg/mL) incubation for 24 hours decreases the viability of NTera-2 cells, and increases caspase-3, -8 and -9 activities, Bax and cytoplasmic cytochrome c levels and decreases Bcl-2 levels. [3] In terms of unstable aberrations, the clastogenic effect of Bleomycin sulfate on ADIPO-P2 cells persists for at least 10 days after exposure. Bleomycin sulfate-induced telomere instability in mammalian cells persists for several generations after exposure. Moreover, the appearance of telomere fusions in Bleomycin sulfate-exposed cells 10 days after treatment suggests that Bleomycin sulfate can induce delayed telomere instability. [4]
In vivo	Day 7 post-Bleomycin sulfate, CD45+ cells in BALf in NOX-/- is 1.7-fold > WT, 57% of which are Mf that decreases by 67% in WT and 83% in NOX-/- by Day 21. [5]
Features

Protocol(Only for Reference)

Cell Assay: ^[4]

Cell lines	ADIPO-P2 cells
Concentrations	2.5 μg/mL
Incubation Time	30 minutes
Method	ADIPO-P2 cells are grown in D-MEM high glucose medium supplemented with 20% fetal calf serum, penicillin (100 U/mL) and streptomycin (100 μg/mL) at 37 °C and 5% CO2 atmosphere. Cells are cultured as monolayer in TC25 Corning flasks containing 1.5 × 105 cells/mL. For each experiment, two flasks are set up, one for the control and one for the treated culture. During the log phase of growth ADIPO-P2 cells are treated with a 30 minutes pulse of 2.5 μg/mL of Bleomycin sulfate. Control cultures are set up in parallel but not exposed to Bleomycin sulfate. Time of exposure and concentration of Bleomycin sulfate are chosen according to previous studies carried out in our laboratory with mammalian cells exposed to Bleomycin sulfate. At the end of the pulse treatment with Bleomycin sulfate, the cells are washed twice with Hank’s balanced salt solution and kept in culture with fresh culture medium until harvesting. Cells are continuously maintained in culture during 5 passages or subcultures after treatment. Subcultivation is carried out whenever the cultures became confluent (approximately 4 × 105 cells/mL of culture medium). To estimate cell growth, at the time of subcultivation cells are collected by trypsinization, an aliquot of about 200 μL stained with 0.4% trypan blue, and the number of viable cells is determined. Cells are then suspended in fresh culture medium and dispensed into new culture flasks containing 1 × 105 cells/mL to continue growing. The rest of the cells is discarded or dispensed in another flask for cytogenetic analysis, which is performed at 18 hours and 10 days after the end of treatments. To analyze chromosomal aberrations, colchicine (0.1 μg/mL) is added to cell cultures during the last 3 hours of culture. Chromosome preparations are made following standard procedures. After harvesting, cells are hypotonically shocked, fixed in methanol:acetic acid (3:1), spread onto glass slides and processed for PNA-FISH. Two independent experiments are carried out.

Animal Study: ^[5]

Animal Models	gp91phox-/- mice, mmp12-/- mice and C57BL6 mice
Dosages	0.075 U/mL
Administration	Administered via i.t.
Solubility	Saline, 30 mg/mL

1

References

Clinical Trial Information( data from http://clinicaltrials.gov)

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT01172860	Recruiting	Colorectal Cancer	Mercy University Hospital, Cork, Ireland|St. James’s Hospital, Dublin|The Adelaide and Meath Hospital	2013-01	Phase 1
NCT01652261	Recruiting	Hodgkin Lymphoma	European Organisation for Research and Treatment of Cancer - EORTC|Polish Lymphoma Research Group	2013-05	Phase 3
NCT01873326	Recruiting	Germ Cell Tumors	Memorial Sloan-Kettering Cancer Center|University of Southern California|Mayo Clinic|University of Pittsburgh|University of North Carolina|University of Chicago|Stanford University	2013-06	Phase 2
NCT01435018	Not yet recruiting	HIV-1 Infection	AIDS Clinical Trials Group|National Institute of Allergy and Infectious Diseases (NIAID)|National Cancer Institute (NCI)|National Institute of Dental and Craniofacial Research (NIDCR)	2013-08	Phase 3
NCT01941901	Recruiting	Cutaneous Metastases	Herlev Hospital	2013-09	Phase 2

Chemical Information

Download Bleomycin Sulfate SDF

Chemical Name

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	1.51262	15.1262	30.2524	45.3786

Customer Reviews (1)

Click to enlarge	Rating
	Source	Bleomycin Sulfate purchased from Selleck
	Method	Colony formation assays
	Cell Lines	HEK293 cell
	Concentrations	0-5 μg/ml
	Incubation Time
	Results	It shows a dose response of killing with increasing concentrations of Bleomycin

Product Citations (1)

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  MCM8-9 complex promotes RAD51 recruitment at DNA damage sites to facilitate homologous recombination. [Park J, et al. Mol Cell Biol 2013;33(8):1632-44]