Ukrain, an alkaloid thiophosphoric acid derivative of Chelidonium majus L. protects human fibroblasts but not human tumour cells in vitro against ionizing radiation.
OBJECTIVE
Ukrain, an alkaloid thiophosphoric acid derivative of Chelidonium majus L., has demonstrated a promising impact on chemotherapy in a variety of malignancies. The effects of the drug on cell survival, alteration of the cell cycle and induction of apoptosis were examined without and in combination with ionizing radiation (IR). The TP53 status of the cell lines used was also investigated.


METHODS
Exponentially growing human tumour cell lines MDA-MB-231 (breast), PA-TU-8902 (pancreas), CCL-221 (colorectal), U-138MG (glioblastoma), and human skin and lung fibroblastic cells, HSF1, HSF2 and CCD32-LU were studied by colony assay, flow cytometry (cell-cycle, annexin-V staining for apoptosis) and Western blotting. Ukrain was used in concentrations from 0.1 to 50 microg ml(-1) for 1, 3 and 24 h and radiation as single doses of 1-10Gy. Combined drug-radiation exposure employed 1 microg ml(-1) Ukrain for 24h plus 2-8 Gy.


RESULTS
Ukrain cytotoxicity was time- and dose-dependent. The combination of Ukrain plus IR gave enhanced toxicity in CCL-221 and U-138MG cells, but not in MDA-MB-231 and PA-TU-8902 cells. Most strikingly, a radioprotective effect was found in normal human skin and lung fibroblasts. Flow-cytometry analyses supported the differential and cell line-specific cytotoxicity of Ukrain. CCL-221 and U-138MG cells accumulated in G2 after 24-h Ukrain treatment, whereas no alterations were detected in the other tumour cells and normal fibroblasts tested. Western blotting of TP53 demonstrated non-functional overexpression in all tumour cell lines without affecting p21. HSF1 presented wild-type TP53 and a p21 response after IR. Flowcytometric analyses of annexin-V staining showed no induction of apoptosis after Ukrain treatment in comparison with untreated controls.


CONCLUSIONS
Differential effects of Ukrain in modulating radiation toxicity of human cancer cell lines and its protective effect in normal human fibroblasts suggest that this alkaloid may have potential properties for clinical radiochemotherapy.
Cordes
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N
Section of Radiobiology and Molecular Environmental Research, Eberhard-Karls-University Tüebingen, Hoppe-Seyler-Strasse 3, 72076 Tübingen, Germany.
Plasswilm
L
L
Bamberg
M
M
Rodemann
H P
HP
D000276:Adjuvants, Immunologic / Q000627:therapeutic use*; D000470:Alkaloids / Q000627:therapeutic use*; D017304:Annexin A5 / Q000494:pharmacology; D017209:Apoptosis; D001600:Berberine Alkaloids; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects / Q000528:radiation effects; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D028281:Chelidonium / Q000378:metabolism*; D017024:Chemotherapy, Adjuvant; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects* / Q000528:radiation effects*; D005434:Flow Cytometry; D006801:Humans; D009369:Neoplasms / Q000188:drug therapy* / Q000473:pathology / Q000532:radiotherapy*; D010617:Phenanthridines; D010710:Phosphates / Q000737:chemistry*; D011837:Radiation-Protective Agents / Q000627:therapeutic use*; D013997:Time Factors; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis
D000276:Adjuvants, Immunologic; D000470:Alkaloids; D017304:Annexin A5; D001600:Berberine Alkaloids; D004791:Enzyme Inhibitors; D010617:Phenanthridines; D010710:Phosphates; D011837:Radiation-Protective Agents; D016159:Tumor Suppressor Protein p53; C060921:ukrain; C035638:thiophosphoric acid