Effect of Bleomycin on Oral Leukoplakia Cell line- an in Vitro Study

Abstract

Background Leukoplakia generally refers to a firmly attached white patch on a mucous membrane which is associated with an increased risk of cancer. So it is important to diagnose leukoplakia in early stage. In this study we are trying to treat leukoplakia as a marker for prediagnosis of cancer. Since, bleomycin as a chemotherapeutic agent, is mainly used in the treatment of multiple tumors, and several cancers. Hence in this study effect of bleomycin was studied in D38, a leukoplakia cell line.Methods Percent lethality values of leukoplakia cells after treated with several concentations of bleomycin was studied by acridine orange/ethidium bromide staining (AO/EB), propidium iodide (PI) staining, 3-(4, 5-dimethythiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay and comet assay. Minimum inhibitory concentration (MIC), lethal concentration25 (LC25), lethal concentration50 (LC50), lethal concentration75 (LC75) and highest permissive concentration (HPC) was calculated from probit computational method.Results From AO/EB staining, PI staining, MTT assay and comet assay the MIC value was 15 mg/L concentration of bleomycin, where as the HPC was found at 100 mg/L of bleomycin. The comet tail length gradually increased from 15 mg/L to 100 mg/L of bleomycin. Percent lethality values were also increased from 15 to 100 mg/L concentration of bleomycin in all the staining protocols as well as in comet assay technique.Conclusion It is important to diagnose leukoplakia in early stage. Since, bleomycin as a chemotherapeutic agent, it is mainly used in the treatment of multiple tumors, testicular carcinomas, lymphomas, and head, neck cancers. From the results, it was found that bleomycin can be used for treatment of oral leukoplakia to prevent its development to SCC.