Klimaszewska Marzenna, Górska Sandra, Dawidowski Maciej, Podsadni Piotr, Szczepanska Agnieszka, Orzechowska Emilia, Kurpios-Piec Dagmara, Grosicka-Maciag Emilia, Rahden-Staroń Iwonna, Turło Jadwiga
Department of Drug Technology and Pharmaceutical Biotechnology, Medical University of Warsaw, Warsaw, Poland.
Department of Biochemistry, Medical University of Warsaw, Warsaw, Poland; Department of Molecular Biology, Institute of Biochemistry, Faculty of Biology, University of Warsaw, Warsaw, Poland.
Int J Med Mushrooms. 2017;19(8):709-716. doi: 10.1615/IntJMedMushrooms.2017021250.
Numerous formulations derived from the shiitake medicinal mushroom, Lentinus edodes, demonstrate anticancer activities. We hypothesized that isolates from selenium (Se)-enriched mycelia of L. edodes would possess stronger cancer-preventive properties than current preparations. The aim of this study was to investigate whether the presence of Se-methyl-seleno-L-cysteine in mycelial extracts of L. edodes affects their cytotoxic activity (makes them stronger) or whether they are as effective as Se-containing polysaccharides. Extracts were prepared from Se-containing mycelia under various conditions and assayed for cytotoxic activity in cancer (PC3 and HeLa) and normal (HMEC-1) cell lines. The chemical composition of the extracts was examined; specifically, the amounts of potentially cytotoxic Se compounds (methylselenocysteine, selenomethionine, and Se-containing polysaccharides) were measured. The relationship between extract composition and biological activity was characterized. Mycelial cultures were cultivated in a 10-L bioreactor in medium enriched with sodium selenite. Mycelial extracts were prepared either at 100°C or at 4°C in acidic solution. Total Se content was determined using the atomic absorption spectrometry method, and methylselenocysteine and selenomethionine contents were measured using reverse-phase high-performance liquid chromatography. Protein, carbohydrate, and polyphenolic contents were determined with spectrophotometric methods, and Se-containing polysaccharides were measured with the use of precipitation. Anticancer activity of mycelial extracts was examined using the MTT cell viability assay. Extracts containing Se-methyl-seleno-L-cysteine or Se-polysaccharides prepared at 4°C and 100°C, respectively, display moderate, time-dependent, specific cytotoxic activity in HeLa and PC3 cell lines. The effect in HeLa cells is more pronounced in the extract prepared at 4°C than at 100°C. The effect is almost equal for the PC3 cell line. However, both extracts have no effect or only slightly stimulate normal (HMEC-1) cell viability. The selective cytotoxic activity of L. edodes extracts in cancer (PC3 and HeLa) cells is due to the presence of both Se-methyl-seleno-L-cysteine and selenated polysaccharides, perhaps in combination with other active ingredients.
源自香菇（Lentinus edodes）的多种制剂均显示出抗癌活性。我们推测，从富硒香菇菌丝体中分离得到的物质可能比目前的制剂具有更强的防癌特性。本研究的目的是调查香菇菌丝体提取物中硒代甲基硒代-L-半胱氨酸的存在是否会影响其细胞毒性活性（使其更强），或者它们是否与含硒多糖一样有效。在各种条件下从含硒菌丝体制备提取物，并在癌症（PC3和HeLa）和正常（HMEC-1）细胞系中测定其细胞毒性活性。检测提取物的化学成分；具体而言，测量潜在细胞毒性硒化合物（甲基硒代半胱氨酸、硒代蛋氨酸和含硒多糖）的含量。表征提取物组成与生物活性之间的关系。在富含亚硒酸钠的培养基中，在10-L生物反应器中培养菌丝体培养物。在100°C或4°C的酸性溶液中制备菌丝体提取物。使用原子吸收光谱法测定总硒含量，使用反相高效液相色谱法测量甲基硒代半胱氨酸和硒代蛋氨酸含量。用分光光度法测定蛋白质、碳水化合物和多酚含量，并用沉淀法测量含硒多糖。使用MTT细胞活力测定法检测菌丝体提取物的抗癌活性。分别在4°C和100°C制备的含有硒代甲基硒代-L-半胱氨酸或硒多糖的提取物在HeLa和PC3细胞系中显示出适度的、时间依赖性的、特异性细胞毒性活性。在HeLa细胞中，4°C制备的提取物比100°C制备的提取物效果更明显。对于PC3细胞系，效果几乎相同。然而，两种提取物对正常（HMEC-1）细胞活力均无影响或仅略有刺激。香菇提取物在癌症（PC3和HeLa）细胞中的选择性细胞毒性活性归因于硒代甲基硒代-L-半胱氨酸和硒化多糖的存在，但可能与其他活性成分结合。
