Cytotoxic activity of extracts of marine sponges from NW Spain on a neuroblastoma cell line

Six species of marine sponges collected at intertidal and sublittoral sites of the coast of Galicia (NW Spain) were screened for potential cytotoxic properties on Neuroblastoma BE(2)-M17 cell line. Exposure to Halichondria panicea, Pachymatisma johnstonia, Ophlitaspongia seriata and Haliclona sp. aqueous extracts strongly affected cell appearance, inducing loss of neuron-like morphology and the formation of clumps. Extracts from these species also caused significant rates of cell detachment and decrease of mitochondrial membrane potential. Incubation with P. johnstonia, O. seriata and Suberites massa extracts also decreased the rate of cell proliferation. The increase of incubation time enhanced propidium iodide uptake by neuroblastoma cells. Toxic responses triggered by sponge extracts are compatible with apoptotic phenomena in neuroblastoma cells, even though increasing propidium uptake at long periods of exposure might indicate the induction of secondary necrosis. The cytotoxic properties of the tested extracts suggest the presence of compounds with potential pharmacological or biotechnological applications in the screened sponge species.     

Cytotoxic activity of methanol extracts from Basidiomycete mushrooms on murine cancer cell lines

Crude methanol extracts of 58 mushroom species were screened for their cytotoxic activities against two murine cancer cell lines, L1210 and 3LL, using the tetrazolium assay. A majority of extracts (74%) exhibited IC50 > 100 microg/ml against both cell lines. A most marked activity against one of the cell lines was noted for nine species (14% of the tested species). While Amanitales and Russulales tested were not found active, Polyporales and Boletales gave better results. Four species exhibited a significant cytotoxic activity (IC50 < or = 20 microg/ml) against at least one of the two murine cancer cell lines (Ganoderma lucidum, Meripilus giganteus, Suillus granulatus, S. luteus). The last one had never been investigated for its cytotoxic compounds before.     

Cytotoxic effects of haplamine and its major metabolites on human cancer cell lines

Haplamine, extracted from Haplophyllum perforatum, is widely used in Central Asia for treating various diseases, including testicular cancer. The purpose of the present study was to investigate in vitro the cytotoxic properties of haplamine and its major metabolites (trans/cis-3,4-dihydroxyhaplamine) on human pancreatic cancer, colorectal cancer and hepatic cancer cell lines. The efficacy of haplamine was compared with those of the respective reference drugs for treating digestive cancers (e. g., 5-FU, gemcitabine). Finally, the implication of apoptosis in haplamine-induced cell death was investigated. The IC50 values of of haplamine were 52.5 +/- 2.6, 24.3 +/- 0.7; 41.5 +/- 2.5, 72 +/- 2, 32 +/- 2.2 and 59.7 +/- 2.1 microM in human pancreatic cancer (Capan1 and Capan2), colorectal cancer (LS174T, HT29, and SW620) and hepatic cancer (HepG2) cells, respectively. The IC50 values of trans/cis-3,4-dihydroxyhaplamine were both > 200 microM, thus suggesting that the previously reported cytotoxic efficacy of haplamine was supported by the parent drug only. Besides, our data showed that haplamine leads to cell death through the induction of early/late apoptosis in the target cells. Interestingly, we found that haplamine showed significant antiproliferative efficacy on resistant SW620 colorectal cells, whereas the reference drug 5-FU was ineffective (32 vs. 73 microM, p < 0.01 t- test), thus suggesting that haplamine could be of interest for treating digestive cancers resistant to standard fluoropyrimidines. Similarly, haplamine proved to be significantly more potent in pancreatic cells than gemcitabine, the reference cytotoxic drug for treating pancreatic carcinomas. Overall, these results confirm the anticancer properties of haplamine suggested by its traditional use, and indicate that it could be further considered in various other solid tumours frequently encountered in adults, including those resistant to standard chemotherapy.     

Cytotoxic effects of four Caryophyllaceae species extracts on macrophage cell lines

Context: Saponins have been reported to possess antitumor properties, to inhibit angiogenesis and to induce tumor apoptosis.

Objective: To test the possible cytotoxic effect of crude extracts from four Caryophyllaceae species including Gypsophila paniculata L., Gypsophila trichotoma Wend., Saponaria officinalis L., and Dianthus sylvestris Wulffen on cultured monocyte/macrophage cell lines.

Materials and methods: After acid hydrolysis of the methanol-aqueous extracts, two representative prosaponins of the Caryophyllaceae, gypsogenin 3-O-glucuronide and quillaic acid 3-O-glucuronide were purified using solid-phase extraction (SPE), then identified by ultra-performance liquid chromatography–electrospray/mass spectrometry (UPLC-ESI/MS). Cytotoxic activity of the crude extracts at concentrations ranging from 0.1 to 200?µg/ml was evaluated on rat alveolar macrophage NR8383 and human monocytic THP-1 cell lines. Apoptosis was determined by measuring caspase-3 activity.

Results: Quantitative analysis by reversed-phase high-performance liquid chromatography (RP-HPLC) revealed a high content of gypsogenin 3-O-glucuronide in Gypsophila species roots (0.52–1.13% dry weight). At a concentration ≥10?µg/ml of crude extracts, a significant reduction of NR8383 and THP-1 cell lines viability was evidenced using the Trypan blue exclusion test. D. sylvestris extract exhibited the highest toxicity against THP-1 cells. Caspase-3 activation was evidenced after 4 and 24?h incubation of macrophages with 100?µg/ml of S. officinalis and G. trichotoma extracts, indicating apoptosis induction.

Discussion and conclusion: Crude extracts from the assayed species revealed cytotoxic effects toward macrophage cell lines. In Gypsophila species, gypsogenin 3-O-glucuronide derivatives could be responsible for the observed cytotoxicity. Therefore, crude extract of Caryophyllaceae is worth investigating for the potential development of agents against cancer cells.     

Cytotoxic and pro-apoptotic activities of cynaropicrin, a sesquiterpene lactone, on the viability of leukocyte cancer cell lines

Cynaropicrin, a sesquiterpene lactone from Saussurea lappa, has been reported to possess immunomodulatory effects on cytokine release, nitric oxide production and immunosuppressive effects. In this study, we have examined cytotoxic effect of cynaropicrin against several types of cell lines such as macrophages, eosinophils, fibroblasts and lymphocytes. Cynaropicrin potently inhibited the proliferation of leukocyte cancer cell lines, such as U937, Eol-1 and Jurkat T cells, but some other cells such as Chang liver cells and human fibroblast cell lines were not strongly suppressed by cynaropicrin treatment. The cytotoxic effect of cynaropicrin was due to inducing apoptosis and cell cycle arrest at G1/S phase, according to flow-cytometric, DNA fragmentation and morphological analyses using U937 cells. Evidence that combination treatment with l-cysteine and N-acetyl-l-cysteine, reactive oxygen species scavengers, or rottlerin (1-[6-[(3-acetyl-2,4,6-trihydroxy-5-methylphenyl)methyl]-5,7-dihydroxy-2, 2-dimethyl-2H-1-benzopyran-8-yl]-3-phenyl-2-propen-1-one), a specific protein kinase (PK) Cdelta inhibitor, abolished cynaropicrin-mediated cytotoxicity and morphological change, and that cynaropicrin-induced proteolytic cleavage of PKCdelta suggests that reactive oxygen species and PKCdelta may play an important role in mediating pro-apoptotic activity by cynaropicrin. Taken together, these results indicate that cynaropicrin may be a potential anticancer agent against some leukocyte cancer cells such as lymphoma or leukemia, through pro-apoptotic activity.     

Cytotoxic effects of novel polyoxotungstates and a platinum compound on human cancer cell lines

The cytotoxicity of a new platinum compound Pt1 [2,9-dimethyl-4,7-diphenyl-1,10-phenanthrolinedichloroplatin(II)] and six polyoxometalates (POM1-6) on two neuroblastoma cell lines (SHEP-SF and KCN) and an Ewing's Sarcoma cell line (CADO-ES-1) was studied. Cisplatin [cis-diamminedichloroplatinum(II)] and carboplatin [cis-diammine(cyclobutanedicarboxylato)platinum(II)] were used as reference agents. Using MTT tests, the cytotoxicity (LD50: lethal doses 50%) of the compounds were measured at different concentrations. After 72 h exposure, the LD50 data for the platinum-containing substances ranged between 4.47 x 10(-6) and 1.91 x 10(-4) M. The SHEP-SF cell line displayed the highest sensitivity to cisplatin. The novel platinum agent Pt1 had a similar cytotoxic effect to the reference agent cisplatin. Both cisplatin and Pt1 were more cytotoxic than carboplatin. The POMs reduced cell viability compared to untreated cells at concentrations between 8.4 x 10(-7) and 3.47 x 10(-5) M. POM1 ([(CH3)4N]2Na6.5(NH4)2[SnII1.5(WO2(OH))0.5(WO2)2(SbW9O33)2] x 32H2O) was the most effective polyoxoanion with a mean LD50 value of 8.83 x 10(-6) M in the three cell lines tested. With CADO-ES-1 and KCN cells, POM1 was found to be more effective than the platinum compounds cisplatin, carboplatin and Pt1.     

Cytotoxic activities of 6-arylamino-7-halo-5,8-quinolinediones against human tumor cell lines

6-Arylamino-7-halo-5,8-quinolinediones (4a-4k, 5a-5b) were tested for in vitro cytotoxicity against human solid tumor cell lines such as A 549 (non-small cell lung), SK-OV-3 (ovarian), SK-MEL-2 (melanoma), HCT-15 (colon) and XF 498 (CNS) by SRB assay. The arylamino-7-chloro-5,8-quinolinediones 4 were also evaluated for cyclin-dependent kinase (CDK2 and CDK4) inhibitory effect. Among them, the 5,8-quinolinediones 4a and 5a with 7-(4-fluorophenyl)amino group were found to be potent cytotoxic against HCT 15, SKOV-3 and XF 498, and the compounds 4f and 4i showed inhibitory activities for the CDK4.     

胜红蓟内生真菌乙酸乙酯提取物的抗氧化及抑菌活性研究

越来越多证据表明,内生真菌代谢产物具有重要的生物学活性,胜红蓟(Ageratum conyzoides)是菊科一种具有多种药理活性的入侵草本植物.我们从胜红蓟根、茎、叶和花中分离并鉴定内生真菌,并对其乙酸乙酯提取物的体外抗氧化活性(DPPH、ABTS+和FRAP测定)、抑菌活性及黄酮、多酚含量进行测定.结果 显示,从胜...     

Antiproliferative effects of extracts from Iranian Artemisia species on cancer cell lines

Objective: Different species of Artemisia (Asteraceae) have shown to exhibit antitumor activity. The aim of this study was to identify the antiproliferative effect of some Artemisia species from Iran on cultured human cancer cells.

Materials and methods: Methanol, ethyl acetate, dichloromethane and n-hexane extracts from aerial parts of seven species of Artemisia were prepared and their antiproliferative effects on four cancer (AGS, HeLa, HT-29 and MCF-7) and normal cell line (L929) were determined. Different concentrations of extracts were added to cultured cells and incubated for 72?h. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was employed to assess the cell viability.

Results: Different extracts exert various growth inhibitory effects. In case of AGS cells, dichloromethane and methanol extracts of A. ciniformis Krasch. & Popov ex Poljak. (IC₅₀ values: 35 and 60 µg/ml, respectively) showed the highest growth inhibitory effects. HeLa cells were more sensitive to both A. diffusa Krasch. ex Poljak. dichloromethane (IC₅₀ value: 71 µg/ml) and A. ciniformis ethylacetate (IC₅₀ value: 73 µg/ml) extracts. Dichloromethane extracts of A. diffusa, A. santolina Schrenk and A. ciniformis (IC₅₀ values: 42, 91 and 94 µg/ml, respectively) exhibited more inhibition on HT-29 cells in comparison to other extracts. MCF-7 cells were best inhibited by A. ciniformis dichloromethane (IC₅₀ value: 29 µg/ml) and A. vulgaris L. ethyl acetate (IC₅₀ value: 57 µg/ml) extracts.

Discussion and conclusion: This study shows the antiproliferative effects of Artemisia extracts on malignant cell lines. Artemisia could be also considered as a promising chemotherapeutic agent in cancer treatment.     

Comparative analysis of protein profiles of aqueous extracts from marine sponges and assessment of cytotoxicity on different mammalian cell types

Marine natural products extracted from sponges represent a new source for drug discovery. Here we describe a simple method for preparing aqueous extracts from 7 Mediterranean demosponges, which allowed the extraction of water-soluble compounds, such as proteins by homogenization of sponge tissue in phosphate buffered saline (PBS).The comparative analysis by SDS-PAGE showed differences in number of bands, bandwidth and intensity among the sponges analyzed. The PAS/silver staining revealed a substantial and different glycoprotein assortment among the demosponges studied.To further study the biological activities present in the sponge extracts, we determined the non-cytotoxic doses on four different mammalian cell types demonstrating that the optimal non-cytotoxic doses were cell type- and extract-dependent.In conclusion, the extraction method described in this paper represents a fast and efficient procedure for the extraction of water-soluble proteins from marine sponges. Furthermore, the cell viability data suggest the feasibility of this method for the direct in vitro cell-based assays.     

Cytotoxic and antitumor activity of liposome-incorporated sclareol against cancer cell lines and human colon cancer xenografts.

The aim of this study was to design and prepare liposome-incorporated sclareol--a highly lipophilic natural product-to overcome its water insolubility and develop suitable formulations for in vivo administration. The bioactive labdane-type diterpene sclareol was incorporated into liposomes composed of egg phosphatidylcholine and dipalmitoylphosphatidylglycerol prepared by the thin-film hydration method followed by sonication. A formulation of egg phosphatidylcholine/dipalmitoylphosphatidylglycerol/sclareol (9:0.1:5 molar ratio) was developed and characterized. The lipid recovery and the sclareol to lipid molar ratio were measured using high-performance thin-layer chromatography/flame ionization detection. In vitro drug release was measured in supplemented RPMI-1640 at 37 degrees C. The liposomal and the free sclareol were initially tested in vitro for their activity against human cancer cell lines using the sulphorhodamine B assay. Liposomes incorporating sclareol at a drug to lipid molar ratio of 0:43, suggesting an incorporation efficiency of almost 80%, showed reduced growth rate of human colon cancer tumors (HCT116) developed in SCID mice, without any significant side effects.     

The effect of culture liquid ethyl acetate mycelium extracts of medicinal mushrooms on the viability of human pancreatic cancer cells

Pancreatic cancer, one of the deadliest of all solid malignancies, is one of the leading causes of cancer death worldwide, with 232,000 new cases and 213,000 deaths reported each year. These unfortunate statistics reflect the advanced stage at which most patients with pancreatic cancer are diagnosed and the paucity of effective chemotherapeutic regimens. Fungal metabolites have been gaining scientific interest because of their medicinal properties. In the present study, 31 different mushroom extracts of 12 medicinal mushroom species were screened for their effect on the viability of human pancreatic adenocarcinoma cells. Extraction procedures were executed with organic solvents--ethanol (EAL), ethyl acetate (EAC), and chloroform (CHL). In some cases, culture liquid (CL) extraction was also performed. All extracts were diluted to a concentration of 50 mg/mL dimethyl sulfoxide. Extract effects on cell viability were examined in human pancreatic adenocarcinoma cells HPAF-II (well differentiated) and PL5 (porrly differentiated), using XTT assay and crystal violet assay (CV). Furthermore, extract effects on LDH leakage were also studied in order to exclude necrotic damage of the extract. The screening phase revealed that among the total 31 extracts examined with various treatment doses (50-500 μg/mL) administered for 72 h, the CL extract of the mushroom Cyathus striatus exhibited the most prominent decrease in cell viability. Moreover, exposure of cells to lower concentrations then the above (1, 2.5, 5, 7.5, 10, 15, 20, and 50 μg/mL) for 24, 48, and 72 h showed a significant decrease in cell viability. Crystal violet results support these findings, and LDH levels measured suggest the lack of a necrotic effect of the extract. Our results indicate that C. striatus CL extract inhibits the viability of human pancreatic adenocarcinoma cells; HPAF-II and PL45. Growth inhibition can be achieved in low concentrations of the extract and a short exposure period. This effect can be mediated through apoptosis induction and/or cell cycle arrest; therefore, additional experiments are needed in order to elucidate the extract mechanism of action. These findings may lead to the development of new therapeutic strategies for the treatment of pancreatic cancer.     

Cytotoxic and antimitotic activities in aqueous extracts of eight cyanobacterial strains isolated from the marine sponge Petrosia ficiformis

Marine cyanobacteria are photosynthetic prokaryotes of significant ecological interest, living free or in association with invertebrates. They are also considered as excellent sources of antineoplastic, antibacterial, antiviral and antifungal compounds. In this work, aqueous extracts from eight cyanobacterial strains isolated from the Mediterranean sponge Petrosia ficiformis have been investigated for their bioactive properties. Bioassays with human erythrocytes, Artemia salina nauplii, and Paracentrotus lividus gametes and embryos were performed. Some aqueous extracts exhibited citolytic effect on human erythrocytes and toxic activity against A. salina nauplii. Furthermore antimitotic activity was evidenced during sea urchin embryos development and disorganization of blastomeres with altered cell-cell contact was also induced. Some of the isolated cyanobacterial strains, belonging to Leptolyngbya and Synechococcus genera with an high citotoxic activity, should be further investigated to better characterize their bioactive molecules. Our data confirm cyanobacteria as an interesting source of novel bioactive compounds with potential applications in pharmaceutics.     

Suvanine analogs from a Coscinoderma sp. marine sponge and their cytotoxicities against human cancer cell lines

Archives of Pharmacal Research - Nine suvanine analogs including suvanine phenethylammonium salt and two new compounds were isolated from the marine sponge Coscinoderma sp., collected from Chuuk...     

Cytotoxic activity of sesquiterpenoids from Atractylodes ovata on leukemia cell lines

The rhizome of Atractylodes ovata (Bai Zhu in Chinese) is a widely used traditional Chinese herb in Taiwan as a tonic agent. In this paper, four sesquiterpenoids, namely atractylon, and atractylenolides I, II, and III, were isolated from the n-hexane extract of A. ovata and were evaluated for cytotoxic effects in vitro. Atractylon significantly inhibited the growth of human leukemia cell line HL-60 and mouse leukemia cell line P-388, and showed low cytotoxicity against primary cultures of normal human peripheral blood mononuclear cells at 15 microg/ml for 12 h. Atractylon had a dose-dependent antiproliferative effect on the two tumor cell lines. In accordance with DNA fragment increases and PARP protein decreases, atractylon at 15 microg/ml for 6 h induced apoptosis in HL-60 cells. Moreover, atractylon inhibited the viability of P-388 cells and induced apoptosis after 15 microg/ml treatment for 12 h in an in vitro assay. However, atractylenolide I at 30 microg/ml for 12 h also induced apoptosis in HL-60 and P-388 cells, but atractylenolides II and III showed no significant inhibition effects on tumor cell growth. As the above results suggested, atractylon and atractylenolide I were the major cytotoxic principle constituents of A. ovata on leukemia cell lines.