Bleomycin sulfate (Blenoxane) is an anticancer agent for squamous cell carcinomas with IC50 of 4 nM in UT-SCC-19A cells. Bleomycin sulfate inhibits DNA synthesis and causes DNA scissions at specific base sequences. Bleomycin sulfate also inhibits tumor angiogenesis. Bleomycin sulfate causes single and double-strand DNA breaks in tumor cells, which interrupts the cell cycle. Bleomycin is thought to achieve this by chelating metal ions, producing a pseudoenzyme that reacts with oxygen to produce superoxide and hydroxide free radicals that cleave DNA. 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. Bleomycin has been used for the treatment of Hodgkin’s lymphoma in combination with doxorubicin, squamous cell carcinomas, testicular cancer, as well as in animal models of pulmonary fibrosis. Bleomycin sulfate plus Rituximab, Doxorubicin, Vinblastine or Dacarbazine has entered phase II clinical trial hodgkin lymphoma.
|Cell lines||MIA PaCa-2 cell|
|Preparation method||Proliferation assay.
The effect of EGCG and BLM on cell proliferation was determined by using TACS 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation assay (Trevigen, Githersburg). The cells (2000 per well) were incubated with EGCG alone, BLM alone or combination of EGCG and BLM, in triplicate in a 96-well plate and then incubated for 2, 4 and 6 days at 37 °C. A MTT solution was added to each well and incubated for 2 h at 37 °C. An extraction buffer (20 % SDS and 50 % dimethylformamide) was added, and the cells were incubated overnight at 37 °C. The absorbance of the cell suspension was measured at 570 nm using a microplate reader (DAS Technologies, Chantilly, VA). This experiment was repeated twice, and the statistical analysis was performed to obtain the final values.
|Concentrations||0, 10 and 20 μM|
|Incubation time||2, 4 and 6 days|
|Animal models||Bleomycin-induced experimental pulmonary fibrosis model|
|Dosages||2 U/kg in a total of 50 μl|
|Administration||intratracheal (i.t.) injection|
|Body Surface Area (m2)||0.007||0.025||0.15||0.05||0.02||0.5|
|Animal A (mg/kg) = Animal B (mg/kg) multiplied by||Animal B Km|
|Animal A Km|
For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.
Inhibitory effect of (-)-epigallocatechin-3-gallate and bleomycin on human pancreatic cancer MiaPaca-2 cell growth.
Bimonte S, et al. Infect Agent Cancer. 2015 Jul 29;10:22. PMID: 26225138.
Targeting sphingosine kinase 1 attenuates bleomycin-induced pulmonary fibrosis.
Huang LS, et al. FASEB J. 2013 Apr;27(4):1749-60. PMID: 23315259.
Characterization of lung stem cell niches in a mouse model of bleomycin-induced fibrosis.
Banerjee ER, et al. Stem Cell Res Ther. 2012 May 29;3(3):21. PMID: 22643035.
Genotoxicity testing of combined treatment with cisplatin, bleomycin, and 5-fluorouracil in somatic cells of Drosophila melanogaster.
Danesi CC, et al. Mutat Res. 2012 Sep 18;747(2):228-33. PMID: 22640881.
Telomere instability is present in the progeny of mammalian cells exposed to bleomycin.
Paviolo NS, et al. Mutat Res. 2012 Jun 1;734(1-2):5-11. PMID: 22564429.
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