Antiproliferative Effect of Urera baccifera Leaves Against Ovarian Carcinoma Cell Line (OVCAR-3)

Abstract Natural products, especially phytochemicals, have been extensively studies and have exhibited important antiproliferative effects. The American native species Urera baccifera (L.) Gaudich. ex Wedd. (Urticaceae) is widely distributed in Brazil, where it is known as urtiga-vermelha or urtigão. The leaves are popularly used as anti-inflammatory, antirheumatic and in the treatment of gastric disorders. However, the antiproliferative potential of this plant against human tumor cells remain to be elucidated. In this study, we evaluated the antiproliferative effects of U. baccifera leaves extracts and fractions against a panel of human tumor cell lines in vitro besides a chemical evaluation of the most active sample by mass spectrometry (ESI-IT-MSn). The hydroalcoholic extract was inactive while dichloromethane extract showed moderate cytostatic activity against ovarian carcinoma cell line (OVCAR-3, GI50 = 1.5 μg/mL). More, the ethyl acetate and n-butanol fractions did not show important activity against tumour cell while the dichloromethane and hexane fractions showed moderate cytostatic activity against ovarian tumor cell line (OVCAR-3, GI50 = 12.7 and 9.4 μg/mL, respectively). Finally, the chemical profile evaluated by mass spectrometry (ESI-IT-MSn) allowed the detection of flavonoids in the HEU and hydroxylated fatty acid in DEU that can explain partially the biological effects observed. This is the first report of the antiproliferative effects of U. baccifera, and DEU has shown potential as a promising source of bioactive compounds.

Antiproliferative effects of pinostrobin and 5,6-dehydrokavain isolated from leaves of Alpinia zerumbet

Abstract Natural products are a major source of drugs for the treatment of cancer. The species Alpinia zerumbet (Pers.) B.L. Burtt & R.M. Sm, Zingiberaceae, is widely distributed in Brazil where it is known as "colônia". The leaves are commonly used in the treatment of hypertension and dyspepsia, however, the effects of A. zerumbet extracts and isolated substances on human cancer cells remain to be elucidated. This study was designed to identify the chemical constituents of hydroalcoholic and dichloromethane extracts from A. zerumbet leaves and to investigate their in vitro antiproliferative activity. The isolated phytochemicals included kaempferol, dihydro-5,6-dehydrokavain, 5,6-dehydrokavain, and pinostrobin. The hydroalcoholic extract inhibited cellular proliferation only at high concentrations, while the dichloromethane extract showed a moderate antiproliferative effect against leukemia and lung tumor cell lines. 5,6-Dehydrokavain showed potent cytostatic activity against glioblastoma cells and a moderate effect on all other tumor cell lines. Pinostrobin showed potent activity against leukemia and breast tumor cell lines and moderate cytostatic effect against ovarian cell. Furthermore, this is the first report on the isolation of kaempferol and pinostrobin from A. zerumbet leaves. Moreover, the purification process described in this study was effective. These results suggest that A. zerumbet leaves are a promising source of anticancer compounds.

Dynamic Contrast-Enhanced MRI Using a Macromolecular MR Contrast Agent (P792): Evaluation of Antivascular Drug Effect in a Rabbit VX2 Liver Tumor Model

OBJECTIVE: To evaluate the utility of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) using macromolecular contrast agent (P792) for assessment of vascular disrupting drug effect in rabbit VX2 liver tumor models. MATERIALS AND METHODS: This study was approved by our Institutional Animal Care and Use Committee. DCE-MRI was performed with 3-T scanner in 13 VX2 liver tumor-bearing rabbits, before, 4 hours after, and 24 hours after administration of vascular disrupting agent (VDA), using gadomelitol (P792, n = 7) or low molecular weight contrast agent (gadoterate meglumine [Gd-DOTA], n = 6). P792 was injected at a of dose 0.05 mmol/kg, while that of Gd-DOTA was 0.2 mmol/kg. DCE-MRI parameters including volume transfer coefficient (K(trans)) and initial area under the gadolinium concentration-time curve until 60 seconds (iAUC) of tumors were compared between the 2 groups at each time point. DCE-MRI parameters were correlated with tumor histopathology. Reproducibility in measurement of DCE-MRI parameters and image quality of source MR were compared between groups. RESULTS: P792 group showed a more prominent decrease in K(trans) and iAUC at 4 hours and 24 hours, as compared to the Gd-DOTA group. Changes in DCE-MRI parameters showed a weak correlation with histologic parameters (necrotic fraction and microvessel density) in both groups. Reproducibility of DCE-MRI parameters and overall image quality was not significantly better in the P792 group, as compared to the Gd-DOTA group. CONCLUSION: Dynamic contrast-enhanced magnetic resonance imaging using a macromolecular contrast agent shows changes of hepatic perfusion more clearly after administration of the VDA. Gadolinium was required at smaller doses than a low molecular contrast agent.

A novel DNA vaccine constructed by heat shock protein 70 and melanoma antigen-encoding gene 3 against tumorigenesis.

Melanoma antigen-encoding gene 3 (MAGE-3) is an ideal candidate for a tumor vaccine although its potency need to be increased. Heat shock proteins (HSPs) represents a potential approach for increasing the potency of DNA vaccines. In the present study, a fusion DNA vaccine composed of Mycobacterium tuberculosis HSP70 and MAGE-3 was constructed and used to immunize C57BL/6 mice against B16 or B16-MAGE-3 tumor cells. The results show that the HSP70-MAGE-3 fusion DNA vaccine enhanced the frequency of MAGE-3-specific cytotoxic T-cells as compared to the MAGE-3 DNA vaccine or the HSP70/MAGE-3 cocktail DNA vaccine (P<0.05). In conclusion, the results indicate that the HSP70-MAGE-3 fusion DNA vaccine can strongly activate MAGE-3 specific cellular immunological reactions and thus significantly inhibit the growth of B16-MAGE-3 tumors, improving the survival of tumor-bearing mice, and the HSP70-MAGE-3 fusion DNA vaccine has a significant therapeutic effect on the tumors that express MAGE-3 antigens.

Effects of Antitumor Agents to the DNA Synthesis of Cecal Mucosa of Mouse / 체질인류학회지

This experiment was performed to evaluate the morphological responses of the cecal mucosa of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of 5-fluorouracil, mitomycin C or adriamycin. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups. In the experimental groups, each mouse was inoculated with 1 x 10(7) Ehrlich carcinoma cells subcutaneous in the inguinal area. From next day, 0.2 mL of saline, 5-fluorouracil (30 mg/kg), mitomycin C (400 microgram/kg) or adriamycin (2 mg/kg) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of anticancer drugs, each mouse was injected with a single dose of 0.7 micro Ci/gm of methyl-3H-thymidine (25Ci/mmol, Amersham Lab, England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. The number of the labeled epithelial cells of the cecal crypts (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and evaluated. On histological study, in the experimental control and mitomycin C-treated mice, general morphology of the cecal mucosae was similar. And in the 5-fluorouracil-treated mice, slightly swelled epithelial cells and expanded lumen of the intestinal crypts were observed. But in the adriamycin-treated groups, slightly disrupted intestinal crypts, a large number of basophilic epithelial cells and the expanded lumen of the intestinal crypts were observed. On autoradiographic study, number of the labeled cells of normal control, experimental control, 5-fluorouracil treated, mitomycin C-treated, or adriamycin-treated groups were 362.2+/-56.12, 350.7+/-71.13, 215.7+/-80.55, 144.2+/-34.60 and 125.0+/-37.45, respectively. In the adriamycin and mitomycin C-treated groups, poorly-labeled cells containing only a few silver grains were observed more frequently than in those of the normal and experimental control groups. From the above results, adriamycin and mitomycin C suppressed the DNA synthesis of the epithelial cells of the cecal mucosa more severely as compared with 5-fluorouracil did. Especially, adriamycin was more harmful than mitomycin C and 5-fluorouracil on the cecal mucosae.

Effects of Antitumor Agents on Ultrastructure of the Splenic White Pulp of the Mouse Implanted with Ehrlich Carcinoma Cells / 대한해부학회지

This experiment was performed to study the morphological responses of the splenic white pulp, the lymphatic tissue of the spleen, of Ehrlich carcinoma cell-implanted mice to three different anticancer drugs (5-fluorouracil, mitomycin C and AG60). Healthy adult ICR mice weighing 20 g each were divided into normal and experimental groups. In the experimental groups, each of mice was inoculated with 1X10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 ml of saline solution, 5-fluorouracil (30 mg/kg), mitomycin C (400 microgram/kg) or AG60 (30 mg/kg, Taerim Pharm. Co., Seoul, Korea) were injected subcutaneously every other day, and animals were sacrificed at 14th day following the f irst injection. Pieces of the tissues were taken from the spleen, and prefixed with phosphate buffered 2.5% paraformaldehyde-1.5% glutaraldehyde solution (pH 7.3) followed by post-fixation with phosphate buffered 1% osmium tetroxide solution (pH 7.3). Fixed tissue blocks were dehydrated, and embedded in araldite mixture. Ultrathin sections stained with uranyl acetate and lead citrate were observed with a JEM 100CX-II electron microscope. In the experimental control group (carcinoma cell-inoculated mouse), splenic white pulp did not show pronounced morphological alterations, but myelin figures were frequently observed in the cytoplasm of some lymphocytes and reticular cells than those of normal control mice. In the AG60 treated group, splenic white pulp did not show specific morphological defect, but nuclear bodies and severe invaginations of the nuclear envelope of the lymphocytes and reticular cells were observed occasionally. In the mitomycin C treated group, myelin figures, severe invaginations of the nuclear envelope, nuclear protrusions, nuclear bodies and interchromatin granules were frequently observed in the lymphocytes and reticular cells of the white pulp. In the 5-f luorouracil treated group, myelin f igures, severe invaginations of the nuclear envelope, nuclear protrusions, nuclear bodies and interchromatin granules were observed more frequently in the lymphocytes and reticular cells of the white pulp, as compared with those of mitomycin C treated mice. From the above results, 5-f luorouracil or mitomycin C may suppress the splenic immune function of cancerinoculated mice, since they suppress the process of differentiation and maturation of splenic lymphocyte and reticular cells, and 5-fluorouracil was more harmful on the spleen than mitomycin C. Whereas AG60 does not affect remarkably the process of differentiation and maturation of lymphocytes and reticular cells in the splenic white pulp.