Aim - leukemia is characterized by uncontrolled marrow cell proliferation and metastatic foci. We investigated the antitumor potential of a nutrient mixture on malignant leukemia P-388 cells. Methods: the nutrient mixture containing lysine, proline, ascorbic acid, green tea extract and other nutrients is formulated to target key pathways in cancer progression. The cells were treated with the mixture, and tested at doses 0, 10, 50, 100, 500 and 1000 mug/ml in triplicates. The effects were evaluated by cell proliferation, Matrigel invasion, cell morphology and apoptosis. The in vivo effect was measured in male nude mice (n = 12) inoculated with P-388 cells. After randomly dividing in two groups, each group was fed regular and the nutrient mixture supplemented diet and the mice were sacrificed after four weeks. The nutrient mixture decreased P-388 cell proliferation at 500 and 1000 mu g/ml. Only 10 % cells were viable at 1000 mu g/ml. Matrigel invasion was significantly inhibited in a dose dependent manner with virtually total inhibition at 1000 mu g/ml. Cell morphological features notably changed with dose increase to 1000 mu g/ml. Analysis of apoptotic cells on live green caspase kit exhibited gradual increase with the increasing dose of the nutrient mixture, and at 1000 mu g/ml 92 % of P-388 cells were in late apoptosis. Tumors in the group of mice supplemented with the nutrient mixture had 50 % lower weight compared to the tumors in control group (p = 0,0105). Histopathologically, both the groups of tumors were similar, yet size of tumors in the group treated with the nutrient mixture was considerably smaller. Conclusion: these results indicate that the nutrient mixture exhibited significant action against multiple targets in P-388 leukemia and may have potential in human leukemia.

Aim - untreated acute promyelocytic leukemia is the most malignant form of acute leukemias, with median surv ival of less than one month. We investigated in vitro and in vivo synergistic effects of a nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea extract, on acute promyelocytic leukemia HL-60 cells. Methods: In vitro, the HL-60 cells were cultured and exposed to NM at doses 0 - 1000 mu g/ml. Cell viability was assessed by Trypan blue dy e exclusion test, matrix metalloproteinases (MMP) expression by gelatinase zymography, invasion through Matrigel and apoptosis by live green Poly Caspase Detection Kit. In vivo studies were carried out in athymic nude mice subcutaneously inoculated with HL-60 cells. In vitro, NM exhibited a dose dependent reduction in cells viability. Zymography revealed matrix MMP-2 and phorbol 12-myristate 13-acetate induced MMP-9 expression. NM inhibited expression of both MMP in a dose dependent manner. Similar step-wise reduction in the Matrigel invasion by HL-60 cells w as also observed by this combination with incremental doses. Gradually increasing doses of NM induced significant apoptosis in HL-60 cells. In vivo, NM inhibited tumor growth by 50 %. Conclusion: the results indicate that NM significantly suppresses tumor growth, decreases cell viability, inhibits MMP expression, Matrigel invasion and induces apoptosis in HL-60 cells.

Індекс рубрикатора НБУВ: Р569.60-52

Рубрики:

Пухлини шкіри та слизових оболонок

Шифр НБУВ: Ж14160 Пошук видання у каталогах НБУВ 

Aim - Fanconi anemia is a rare genetic disorder with high propensity for development of cancers, such as aplastic anemia, leukemia and head and neck cancers. Collagen digesting matrix metalloproteinase (MMP) enzymes have been implicated in for their role in various malignancies and to promote metastasis. Biological agents, that prevent extracellular matrix digestion by the MMPs, have been shown to be promising therapeutic approaches to cancer. In this study we investigated effects of a nutrient mixture (NM) containing, ascorbic acid, lysine, proline and green tea extract, on human FANCA and FANCC lymphoblasts for viability, MMP secretion and invasion. Human FANCA lymphoblasts GM13022 and HCS536 were challenged with NM at concentration range within 10 - 1000 mu g/ml. Cell toxicity was assessed by Trypan blue dye exclusion test. Invasion was evaluated through Matrigel and gelatinase zymography for MMP activity. NM was toxic in dose dependent mode to HCS536 cells but not to GM13022 cells. GM13022 cells but not HCS536 cells exhibited MMP-9 secretion, which was inhibited by NM. Matrigel invasion was inhibited in HCS536 cells at 100 and 500 mu g/ml by 27 % and 93 %, respectively. In GM13022 cells, the NM showed completely blocked Matrigel invasion at 500 mu g/ml. Conclusion: NM inhibited MMP secretion and Matrigel invasion in FANCA and inhibited invasion and induced toxicity in FANCC lymphoblasts. These results suggest that the NM may have therapeutic potential in Fanconi anemia associated neoplasia.

Aim - Fanconi Anemia, an autosomal recessive disorder, is characterized by chromosomal abnormality leading to birth defects, progressive bone marrow failure, and a high probability of developing malignancy at an early age. Head and neck squamous cell carcinoma and myeloid leukemia are the major causes of cancer related morbidity and mortality in Fanconi anemia patients. We investigated the effect of a nutrient mixture on Fanconi Anemia human fibroblast cell lines FA-A:PD20 and FA-A:PD220 on matrix metalloproteinase expression, invasion, cell proliferation, morphology and apoptosis. The cell lines were grown in a modified Dulbecco's Eagle medium and at near confluence were treated with the nutrient mixture at increasing doses: 0; 10; 50; 100; 500; 1000 mu g/ml. The cells were also treated with PMA to induce MMP-9 expression. Results: Zymography demonstrated MMP-2 and PMA-induced MMP-9 activity. The nutrient mixture inhibited expression of both, MMP-2 and MMP-9, in a dose dependent manner with virtually total inhibition observed at 500 mu g/ml. Matrigel invasion was inhibited in both cells lines; with 100 % inhibition for FA-A:PD20 at 500 mu g/ml and 100 % inhibition of FA-A:P220 cells at 100 mu g/ml. H&E staining did not indicate any change in cell morphology and causes apoptosis at higher doses. Conclusion: our data demonstrated that the nutrient mixture inhibited matrix metalloproteinase expression, invasion and induced apoptosis, the important parameters for cancer prevention. The results suggest that the nutrient mixture may have therapeutic potential in Fanconi Anemia associated neoplasia.

Aim - a nutrient mixture (NM) containing ascorbic acid, lysine, proline and green tea extract has exhibited anticancer activity in vitro and in vivo in a number of cancer cell lines. We investigated the effect of NM on human leukemic myeloid U-937 cells in vitro by measuring: cell proliferation, MMP expression, invasion, apoptosis, and COX-2 and COX-1 protein expression. Human leukemic cell line U-937 (ATCC) was cultured in RPMI medium supplemented with fetal bovine serum and antibiotics. After 24 h, the cells were treated with NM at 0, 50, 100, 250, 500 and 1000 mu g/ml, in triplicate at each dose. Phorbol 12-myristate 13-acetate (PMA), 100 mu g/ml was added to cells to induce MMP-9 secretion. Cell proliferation was evaluated by MIT assay, MMP expression by gelatinase zymography, invasion through Matrigel, apoptosis by using live green caspase detection kit (Molecular Probe), and COX-2 and COX-1 expression by Western blot. NM had no effect on U-937 cell growth at a concentration of 250 mu g/ml and exhibited an antiproliferative effect at 500 mu g/ml concentration. Zymography did not demonstrate MMP-2 or MMP-9 secretion in normal cells; however, PMA strongly induced MMP-9, which was inhibited by NM in a dose-dependent manner. Cell penetration through Matrigel was significantly reduced (by 95 %) at 250 mu g/ml NM and completely blocked at 500 mu g/ml NM. NM induced slight apoptosis at 100 mu g/ml and moderate at 500 and 1000 pg/ml concentration. NM inhibited COX-2 expression in a dose-dependent fashion and had no effect on COX-1 expression. Conclusions: our results suggest that NM has potent inhibitory effects on U-937 cell growth and expression of inflammatory mediators, significant parameters in AML progression.

Індекс рубрикатора НБУВ: Р569.627.7

Рубрики:

Пухлини головного мозку та його оболонок

Шифр НБУВ: Ж14160 Пошук видання у каталогах НБУВ 

Індекс рубрикатора НБУВ: Р733.562,2 + Р733.569.402

Шифр НБУВ: Ж14160 Пошук видання у каталогах НБУВ