Investigation of therapeutic efficiency of bleomycin-glucuronide labeled with radioactive Iodine-131 in the cell level

Bleomycin (BLM) which is used as an anti-cancer agent, was first isolated byJapanese scientist Umezawa as a fermentation product of Streptomyces verticillusand BLM is an important antibiotic which is in glycopeptide form. There are twotypes of BLM. These are BLM A2 and BLM B2. Cytotoxic effect of both arises fromthe ability of causing DNA fragmentation. Effect spectrum is quite wide. BLM has atherapeutic potential in diseases such as lymphocyte cancer, platy ephiteliacarcinoma, testicle cancer and bottom wart.The aim of our study is to radiolabeling Bleomycin (BLM) and Bleomycinglucuronide(BLMG) which is enyzmatically synthesized enyzmatically with 131I andto investigate the incorporation of therapeutical activity of radiolabeled compoundson PC-3 (prostate), Caco (large intestine), Hutu (small intestine) and A549 (lung) oncancerous cell lines in vitro and measure the binding yield of radiolabeledcompounds on cancerous cells. Thus, firstly UDP-glucuronyl transferase enzymewas isolated from rat liver and then, BLMG was enyzmatically synthesized usingBLM and UDPGT and isolation studies of BLMGwas carried out by using HPLCmethod. In the latter step, quality-control studies of BLM and BLMG labeled with 131Iwere carried out by using radiochromatographic method. In this context, it was foundthat the binding yields were obtained to be % 90 and % 80 for 131I-BLM and 131IBLMGrespectively. Subsequently, cell culture studies were carried out and bindingyields of 131I, 131I-BLM ve 131I-BLMG on PC-3 (prostate) cancerous cells wereinvestigated. Optimum time, the amount of matter and specific activity parameterswere determined. With using the optimum conditions, the binding yields of 131I, 131IBLMve 131I-BLM on Caco, Hutu and A549 cancerous cell lines were investigatedand then, all of the experimental data were evaluated and interpreted by usingGraphPAd statistical program. Besides, cytotoxicities of BLM and BLMG on PC-3cancerous cell line were investigated and BLM, BLMG were labeled with FITC.Fluorescent images of BLM and BLMG were taken by focused on 100x and 10x,respectively.In conlusion when uptakes of radiolabeled compounds on all cell lines arecompared, it is found that uptake of 131I-BLMG on cells are 5-6 times higher thanuptake of 131I-BLM. This result shows that BLMG has a much more efficient antitumuoragent potential than that of BLM used as a conventional chemotherapyagent.