Cytological and cytogenetic alterations of human blood T-lymphocytesin vitro induced by anti-cancer drug Adriamycin in combination with shiitake mushroom mycelium (Lentinula edodes) extract

Protective effects of shiitake mushroom mycelium (Lentinulaedodes) extract grown in darkness for 10 days against cytogenetic damage induced by anti-cancer drug Adriamycin (Doxorubicin) wereassessed in vitro in the human peripheral blood T-lymphocyte cultures. It was found that Adriamycin in the cultured lymphocytes caninduce various types of karyopathological alterations including the deceasing frequency of cells with round-shaped nuclei normal forT-lymphocytes and the increasing frequency of micronucleated or protrusion-nucleated cells as well as the cells with apoptosis featureslike karyopyknosis, karyorhexis or karyolysis. Frequency of binucleated cells has also increased in the Adriamycin-exposed cultures.Alternatively, shiitake mycelium extracts did not result in any types of karyopathological alterations in the treated cultures, but inducedthe significant increase in the frequency of cells with bean-shaped nuclei and the decrease in the frequency of cells with oval-shapednuclei. An addition of Adriamycin to the preliminary shiitake extract-treated lymphocyte cultures resulted in the significant diminutionof the karyopathologically alternated cells including micronucleated, karyopyknotic and karyolytic cells as compared to the only Adriamycin-exposed cultures. However, the frequencies of binucleated and protrusion-nucleated cells in the Adriamycin and shiitake combinedlytreated cultures remained high and unchanged as compared to the only Adriamycin-exposed cultures. Mitotic pathology analysisin the Adriamycin-exposed lymphocyte cultures has demonstrated various alterations in more than 40% of the cell divisions. Among thetypes of the mitotic pathology, the anaphase bridges, metaphase chromosome lagging or multi-clustered metaphases have dominated.However, any increases in the frequency of C-mitoses were not detected compared to the intact control. It was found that Adriamycinsignificantly depressed the mitotic activity and blast-transformation of the treated cultures, whereas shiitake mycelium extracts doubledthe mitotic activity and increased blast-transformation as compared to the intact control. Moreover, Adriamycin induced the significantdecrease in the area of the cultured cells, whereas addition of shiitake mycelium extracts to the lymphocyte cultures resulted in the considerableincrease in the frequency of cells with the area more than 100 μm2.

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