Cytotoxic effect of Alpinia scabra (Blume) Náves extracts on human breast and ovarian cancer cells

BACKGROUND: Alpinia scabra, locally known as 'Lengkuas raya’, is an aromatic, perennial and rhizomatous herb from the family Zingiberaceae. It is a wild species which grows largely on mountains at moderate elevations in Peninsular Malaysia, but it can also survive in the lowlands like in the states of Terengganu and Northern Johor. The present study reports the cytotoxic potential of A. scabra extracts from different parts of the plant. METHODS: The experimental approach in the present study was based on a bioassay-guided fractionation. The crude methanol and fractionated extracts (hexane, chloroform and water) from different parts of A. scabra (leaves, rhizomes, roots and pseudo stems) were prepared prior to the cytotoxicity evaluation against human ovarian (SKOV-3) and hormone-dependent breast (MCF7) carcinoma cells. The identified cytotoxic extracts were then subjected to chemical investigations in order to identify the active ingredients. A normal human lung fibroblast cell line (MRC-5) was used to determine the specificity for cancerous cells. The cytotoxic extracts and fractions were also subjected to morphological assessment, DNA fragmentation analysis and DAPI nuclear staining. RESULTS: The leaf (hexane and chloroform) and rhizome (chloroform) extracts showed high inhibitory effect against the tested cells. Ten fractions (LC1-LC10) were yielded after purification of the leaf chloroform extract. Fraction LC4 which showed excellent cytotoxic activity was further purified and resulted in 17 sub-fractions (VLC1-VLC17). Sub-fraction VLC9 showed excellent cytotoxicity against MCF7 and SKOV-3 cells but not toxic against normal MRC-5 cells. Meanwhile, eighteen fractions (RC1-RC18) were obtained after purification of the rhizome chloroform extract, of which fraction RC5 showed cytotoxicity against SKOV-3 cells with high selectivity index. There were marked morphological changes when observed using phase-contrast inverted microscope, DAPI nuclear staining and also DNA fragmentations in MCF7 and SKOV-3 cells after treatment with the cytotoxic extracts and fractions which were indicative of cell apoptosis. Methyl palmitate and methyl stearate were identified in the hexane leaf extract by GC-MS analysis. CONCLUSIONS: The data obtained from the current study demonstrated that the cell death induced by cytotoxic extracts and fractions of A. scabra may be due to apoptosis induction which was characterized by apoptotic morphological changes and DNA fragmentation. The active ingredients in the leaf sub-fraction VLC9 and rhizome fraction RC5 may lead to valuable compounds that have the ability to kill cancer cells but not normal cells

Synthesis of a DNA-targeting nickel (II) complex with testosterone thiosemicarbazone which exhibits selective cytotoxicity towards human prostate cancer cells (LNCaP)

Testosterone thiosemicarbazone, L and its nickel (II) complex 1 were synthesized and characterized by using FTIR, CHN, (1)H NMR, and X-ray crystallography. X-ray diffraction study confirmed the formation of L from condensation of testosterone and thiosemicarbazide. Mononuclear complex 1 is coordinated to two Schiff base ligands via two imine nitrogens and two tautomeric thiol sulfurs. The cytotoxicity of both compounds was investigated via MTT assay with cisplatin as positive reference standard. L is more potent towards androgen-dependent LNCaP (prostate) and HCT 116 (colon). On the other hand, complex 1, which is in a distorted square planar environment with L acting as a bidentate NS-donor ligand, is capable of inhibiting the growth of all the cancer cell lines tested, including PC-3 (prostate). It is noteworthy that both compounds are less toxic towards human colon cell CCD-18Co. The intrinsic DNA binding constant (Kb) of both compounds were evaluated via UV-Vis spectrophotometry. Both compounds showed Kb values which are comparable to the reported Kb value of typical classical intercalator such as ethidium bromide. The binding constant of the complex is almost double compared with ligand L. Both compounds were unable to inhibit the action topoisomerase I, which is the common target in cancer treatment (especially colon cancer). This suggest a topoisomerase I independent-cell death mechanism

Artocarpus heterophyllus Lam. stem bark inhibits melanogenesis through regulation of ROS, cAMP, and MAPK Pathways

Natural-based skin-lightening cosmeceutical products are attracting high popularity nowadays due to their relatively high bioavailability upon application. Artocarpus species have been highlighted with such potential, and our previous studies have reported that Artocarpus heterophyllus Lam. stem bark extract exhibited a potent anti-melanogenic activity by reducing melanin content and inhibiting cellular tyrosinase activity in B16F10 melanoma cells. Hence, this study aimed to identify the bioactive fraction from A. heterophyllus Lam. stem bark and determine its anti-melanogenic mechanisms in B16F10 melanoma cells. Our results showed that a fraction (H-3) demonstrated the most pronounced anti-melanogenic effect at 12.00 µg/mL by reducing melanin content to 22.86 ± 2.90% and inhibiting cellular tyrosinase activity at treatment concentration 33-fold lower than kojic acid, without being cytotoxic against B16F10 melanoma cells. Moreover, treatment with H-3 for 24 and 48 h substantially scavenged intracellular reactive oxygen species (ROS) of hydrogen peroxide-challenged B16F10 melanoma cells by 1.8 and 4.4%, respectively. Based on the microarray profiling and qPCR analysis, H-3 downregulated Creb3l1, Creb3l2, Creb3l3, Mitf, Tyr, Tyrp1, and Dct genes in B16F10 melanoma cells, whereas the expression of Map3k20, Mapk14 (p38), and Foxo3 genes was markedly increased. Altogether, these results demonstrated that H-3 exhibited its anti-melanogenic activity in B16F10 melanoma cells through scavenging ROS and concurrent inhibition of the cAMP and activation of the p38/MAPK signaling pathways. These findings indicate that H-3 has the potential to be used as a skin lightening cosmeceutical agent in the treatment of skin hyperpigmentation

Association analyses of East Asian individuals and trans-ancestry analyses with European individuals reveal new loci associated with cholesterol and triglyceride levels

Large-scale meta-analyses of genome-wide association studies (GWAS) have identified >175 loci associated with fasting cholesterol levels, including total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and triglycerides (TG). With differences in linkage disequilibrium (LD) structure and allele frequencies between ancestry groups, studies in additional large samples may detect new associations. We conducted staged GWAS meta-analyses in up to 69,414 East Asian individuals from 24 studies with participants from Japan, the Philippines, Korea, China, Singapore, and Taiwan. These meta-analyses identified (P < 5 × 10-8) three novel loci associated with HDL-C near CD163-APOBEC1 (P = 7.4 × 10-9), NCOA2 (P = 1.6 × 10-8), and NID2-PTGDR (P = 4.2 × 10-8), and one novel locus associated with TG near WDR11-FGFR2 (P = 2.7 × 10-10). Conditional analyses identified a second signal near CD163-APOBEC1. We then combined results from the East Asian meta-analysis with association results from up to 187,365 European individuals from the Global Lipids Genetics Consortium in a trans-ancestry meta-analysis. This analysis identified (log10Bayes Factor ≥6.1) eight additional novel lipid loci. Among the twelve total loci identified, the index variants at eight loci have demonstrated at least nominal significance with other metabolic traits in prior studies, and two loci exhibited coincident eQTLs (P < 1 × 10-5) in subcutaneous adipose tissue for BPTF and PDGFC. Taken together, these analyses identified multiple novel lipid loci, providing new potential therapeutic targets

The genetics of blood pressure regulation and its target organs from association studies in 342,415 individuals

To dissect the genetic architecture of blood pressure and assess effects on target-organ damage, we analyzed 128,272 SNPs from targeted and genome-wide arrays in 201,529 individuals of European ancestry and genotypes from an additional 140,886 individuals were used for validation. We identified 66 blood pressure loci, of which 17 were novel and 15 harbored multiple distinct association signals. The 66 index SNPs were enriched for cis-regulatory elements, particularly in vascular endothelial cells, consistent with a primary role in blood pressure control through modulation of vascular tone across multiple tissues. The 66 index SNPs combined in a risk score showed comparable effects in 64,421 individuals of non-European descent. The 66-SNP blood pressure risk score was significantly associated with target-organ damage in multiple tissues, with minor effects in the kidney. Our findings expand current knowledge of blood pressure pathways and highlight tissues beyond the classic renal system in blood pressure regulation

Bioactivity and chemical investigations of Pereskia Bleo and Pereskia Grandifollia / Sim Kae Shin

Ethnopharmacological data has been one of the common useful criteria in drug discovery. Pereskia bleo and Pereskia grandifolia (Cactaceae), commonly known as ‘Jarum Tujuh Bilah’ in Malay and ‘Cak Sing Cam’ in Chinese have long been used as natural remedies in Malaysia. The experimental approach in the present study was based on bioassay-guided fractionation. The crude methanol and fractionated extracts of both Pereskia spp. were initially investigated for their biological activities such as antioxidant, antimicrobial and cytotoxic effect against five human cancer cell lines, namely nasopharyngeal epidermoid carcinoma cell line (KB), cervical carcinoma cell line (CasKi), colon carcinoma cell line (HCT 116), hormone-dependent breast carcinoma cell line (MCF7), lung carcinoma cell line (A549) and the non-cancer human fibroblast cell line (MRC-5) using in vitro cytotoxicity assay, in order to identify the bioactive extracts of both Pereskia spp.The hexane and ethyl acetate extracts of both Pereskia spp. generally showed stronger antioxidant activities than the other extracts. Ethyl acetate extracts of both Pereskia spp. also showed some mild antimicrobial activities against the tested bacteria.In the cytotoxicity assay, both Pereskia spp. exerted no damage to MRC-5 normal cells.The ethyl acetate extracts of both Pereskia spp. in general gave higher inhibition and stimulation values against various cancerous cell lines compared to other extracts. The cell deaths of the selected cancer cells elicited by the cytotoxic active extracts of both Pereskia spp. were found to be apoptotic in nature based on a clear indication of DNA fragmentation, which is a hallmark of apoptosis. In addition, the LUX RT-qPCR [realiii time reverse transcriptase–polymerase chain reaction (RT-qPCR) using LUX (Light Upon eXtension) primers] analysis showed that apoptosis elicited by the cytotoxic extracts on selected cancer cells was mediated by p53, caspase-3 and c-myc activation in different expression levels. Methyl palmitate, methyl linoleate, methyl α-linolenate and phytol were identified from the hexane extract of Pereskia bleo by GCMS analysis whilst methyl palmitate, methyl linoleate, methyl α-linolenate and methyl stearate were identified from the hexane extract of Pereskia grandifolia. From the results of the biological screenings, it is observed that the ethyl acetate extracts generally have stronger biological activities than other extracts. Further chemical investigations were thus directed to the ethyl acetate extracts of both Pereskia spp.2,4-Di-tert-butylphenol (1), α-tocopherol (2), phytol (3), ß-sitosterol (4),dihydroactinidiolide (5) and a mixture of sterols containing campesterol (6), stigmasterol (7) and β-sitosterol (4) were isolated from Pereskia bleo whilst 2,4-di-tertbutylphenol (1), α-tocopherol (2), ß-sitosterol (4) and mixture containing methyl palmitate (9), methyl oleate (10), methyl stearate (11) and 2,4-di-tert-butylphenol (1) were isolated from Pereskia grandifolia. It is interesting to note that 2,4-ditertbutylphenol(1), α-tocopherol (2) and ß-sitosterol (4) were isolated from the ethyl acetate extracts of both Pereskia spp. The cytotoxic activities of the isolated constituents were evaluated against the above human cell lines and further studies on their mode of action suggested that these activities are connected with induction of apoptosis.In addition, the toxicity of both Pereskia spp. was evaluated in vivo and were considered safe in acute oral toxicities in experimental mice. The screening of the locally grown Pereskia bleo and Pereskia grandifolia indicated the presence ofalkaloids, but the concentration was very low. The findings of Pereskia bleo and Pereskia grandifolia in the present study provided scientific validation on the use of the leaves of both Pereskia spp. for the treatment of cancer. Further studies on the mutagenic and toxicity effect over a longer period of time involving detection of effects on vital organ functions should be carried out to ensure that the plants are safe for human consumption

Antimelanogenesis and Anti-Inflammatory Activity of Selected Culinary-Medicinal Mushrooms

Five culinary-medicinal mushrooms are commonly available in the Malaysian market: Agaricus bispo-rus (white and brown), Ganoderma lucidum, Hypsizygus marmoreus, Pleurotusfloridanus, and P. pulmonarius. These species were selected for use in the current study, the aim of which was to investigate the antimelanogenesis and anti-inflammatory activity ofthese mushrooms in an attempt to evaluate their potential use in cosmeceuticals. Mushroom fruiting bodies were extracted with hot water, and the extracts were freeze-dried before testing. The antimelanogenesis activity of the extracts was determined by cell viability assay, measurement of intracellular melanin content, and cellular tyrosinase assay with B16F10 melanoma cells. The anti-inflammatory activity of the mushroom extracts was tested by measuring the levels of nitric oxide (NO), tumor necrosis factor (TNF)-a, and interleukin-10 excreted by RAW264.7 macrophages. Brown A. bisporus reduced intracellular melanin content to the largest extent-up to 57.05 ± 3.90%-without a cytotoxic effect on B16F10 melanoma cells. This extract also reduced cellular tyrosinase activity to 17.93 ± 2.65%, performing better than kojic acid, the positive control. In parallel, the extract from brown A. bisporus, at the highest concentration tested, has appreciable anti-inflammatory activity through reductions ofNO and TNF-a levels. The other 5 extracts showed moderate antimelanogenesis and anti-inflammatory activities. In summary, our findings show that A. bisporus (brown) extract has the potential to be used as an ingredient in whitening skincare products and to sooth the inflammatory response on the skin

Evaluation of antioxidant properties, cytotoxicity and acute oral toxicity of Gynura procumbens (Compositae)

Gynura procumbens which is locally known as ‘Sambung nyawa’ in Malay and ‘Feng Wei Jian’ in Chinese, belongs to the botanical family of Compositae. In this study, antioxidant property of G. procumbens extracts was evaluated using DPPH radical scavenging, metal chelating and β-carotene bleaching assays. To the best of our knowledge, this is the first report that evaluated the cytotoxicity of G. procumbens extracts on human colon cancer cells (HT-29, HCT 116, HCT-15, SW480, Caco-2) and human normal colon cells (CCD-18Co). The results showed that ethyl acetate extract contained the highest total phenolic content (172.68 mg of GAEs/g of extract) compared to methanol, hexane and water extracts. Methanol extract possessed better overall antioxidant activities while ethyl acetate extract demonstrated better cytotoxic activity. At 24 h treatment, ethyl acetate extract demonstrated selective cytotoxicity against HT-29 and HCT 116 cells with IC50 values of 35.7 and 42.6 µg/mL, respectively. In addition, methanol extract showed negligible level of toxicity when administered orally. All the results indicated that G. procumbens may provide benefits in prevention and treatment of cancer

Caffeoylquinic acids induce cell death and cell cycle arrest on HCT 116 cells via formation of extracellular H2O2and quinones

Caffeoylquinic acids are frequently present in variety of medicinal plants, fruits, vegetables and beverages. These phenolic acids are widely accepted as antioxidants but have pro-oxidant activity under certain condition. The cell death and cell cycle arrest on HCT 116 human colon carcinoma cells were observed by flow cytometry when treated with 5-O-caffeoylquinic acid (CQ) and 3,5-dicaffeoylquinic acid (DCQ) at 1 and 2 mg/ml. The addition of cell impermeable catalase and reduced glutathione (GSH) could protect HCT 116 cells from cell death and cell cycle arrest effect of CQ and DCQ. It was observed that CQ and DCQ generated extracellular hydrogen peroxide (H2O2) and quinones in the culture medium. These in vitro data showed that caffeoylquinic acids have to undergo oxidative mechanism in order to induce anti-proliferation effect on colon cancer cells and this may be beneficial in prevention or treatment of colorectal cancer