Extracts of edible, medicinal Thai plants inhibit the human breast cancer cells

Purpose: To evaluate the effects of ten edible, medicinal Thai plant extracts on MCF-7 cell viability and cell migration, as well as their mechanism(s) of action. Methods: Ethanolic plant extracts of ten edible, medicinal plants were tested for their cytotoxicity against MCF-7 cells using sulforhodamine B (SRB). To investigate the cytotoxic mechanism(s) of action of these extracts, the study was examined gene expression and protein expression by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. Cell migration was studied by wound healing assay. Results: Four of the ten test extracts were potently cytotoxic, Careya sphaerica (CS), Azadirachta indica (AI), Piper nigrum (PN) and Oroxylum indicum (OI) with half maximal inhibitory concentrations (IC50) less than 100 μg/mL. All four extracts stimulated ROS overgeneration, increased caspase 3 activity and decreased growth-related gene expression including cdk2, cdk4, cdk6, cyclin D1 and cyclin E. Furthermore, the extracts significantly enhanced cyclin-dependent kinase inhibitor (CDKI) p21 levels and activated cancer cell death. The four extracts, CS, AI, PN and OI, also significantly reduced cancer cell migration, with PN being the most potent. Conclusion: Extract of the edible plants CS, AI, PN and OI have in vitro anticancer activity and are promising starting points for the development of breast cancer drugs. Keywords: Careya sphaerica (CS), Azadirachta indica (AI), Piper nigrum (PN), Oroxylum indicum (OI), Breast cancer, Cell deat

Alendronate blocks human cholangiocarcinoma cell proliferation and migration

Purpose: To explore the effect of alendronate on cell death and migration of cholangiocarcinoma (CCA). Methods: Migration and cell death of CCA cells were determined using sulforhodamine B (SRB), colony formation, wound healing, and gelatin zymography assays. The mechanism of action of alendronate was studied with reverse-transcriptase polymerase reaction (RT-PCR) for gene expression and by Western blotting analysis for protein expression. Results: Alendronate stimulated KKU-100 cell death in dose- and time-dependent manner, with low IC50 value, and significantly inhbited colony formation at doses of 5 - 100 µM. Moreover, alendronate at doses of 250 - 1000 µM significantly stimulated CCA apoptosis via reactive oxygen species (ROS) generation, and enhanced caspase 3 activity at a dose of 1000 µM. Moreover, at a dose of 250 µM, it significantly inhibited cell growth through induction of caspase 3 and p53, and reduction of protein expression levels of NF-ĸB. Furthemore, alendronate altered mevalonate (MVA) pathway via downregulation of Rac1 protein expression. In contrast, it significantly inhibited CCA cell migration, and reduced MMP 2 and MMP 9 levels at doses of 25 - 100 µM. Conclusion: Alendronate may be useful as a novel drug for prevention and chemotherapy of CCA

Effects of mycelial extract and crude protein of the medicinal mushroom, Ophiocordyceps sobolifera, on the pathogenic fungus, Candida albicans

Purpose: To investigate the antifungal effect of the mycelial extracts and crude proteins of the medicinal mushroom, Ophiocordyceps sobolifera on Candida albicans.Methods: The antifungal activities of the mycelial extracts and crude proteins of seven strains of the entomopathogenic fungus, Ophiocordyceps sobolifera, were screened against Candida albicans strain NCYC854 using an agar well diffusion method. Minimum inhibitory concentrations (MIC) and minimum fungicidal concentration (MFC) were determined using broth microdilution method. The kinetics of fungal death was elucidated via time-kill assays, and while ultrastructural alteration changes to fungal cells were investigated by scanning electron microscopy (SEM).Results: The antifungal activities of the mycelial extracts were superior to those of the crude proteins. Among the isolates, Cod-KK1643 exhibited the highest activity with the lowest MIC against the test strains. Therefore, it was chosen for further investigations. The isolate Cod-KK1643 exhibited concentration- and time-dependent fungistatic activity in the time kill assay. However, these activities were absent in the crude protein of the isolate. Moreover, Cod-KK1643 mycelial extract induced morphological alterations in fungal cells, such as decreased cell size, and crushed or cracked appearance. Slight alterations in cell morphology (decreased cell size or crushed appearance) were observed in the crude protein treatment.Conclusion: The mycelial extracts of the fungus, O. sobolifera (especially isolate Cod-KK1643) exert potent antifungal activity against human pathogenic fungus, C. albicans.Keywords: Anti-fungal activity, Candida albicans, Entomopathogenic fungus, Ophiocordyceps sobolifer

Antibacterial and anti-breast cancer cell line activities of Sanghuangporus sp.1 extracts

Purpose: To evaluate the activity of extracts of Sanghuangporus sp.1 fungus against pathogenic bacteria and a breast cancer cell line.Methods: The wild fruiting body and mycelium of Sanghuangporus sp.1 were extracted with water and ethanol by ultrasonication extraction. The activity of the extracts against pathogenic bacteria was determined by agar-well diffusion method while minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were obtained by agar-well diffusion and broth macrodilution methods. The cytotoxicity of the extract against human breast cancer cell line MCF-7 was determined by sulforhodamine B assay.Results: Only the ethanol mycelial extract exhibited antibacterial activity. Activity was detected against 6 of the 17 test strains of bacterial pathogen. The MICs and MBCs against these 6 strains were quite low, especially for B. cereus ATCC 11778 (2.5 and 2.5 mg/mL) and S. aureus (MSSA, DMST 2933, 2.5 and 5.0 mg/mL). The ethanol mycelial extract was a more potently cytotoxic against MCF-7 cells than either the aqueous mycelial extract or the ethanol wild fruiting body extract,  inhibiting cell growth at a concentration of 250 μg/mL. The aqueous wild fruiting body extract was inactive against MCF-7 cells when compared with untreated control groups.Conclusion: The ethanol mycelial extract of the medicinal mushroom  Sanghuangporus sp.1, obtained by ultrasonication extraction method, exhibited potent antibacterial and anticancer activity and seems to be a substitute for wild Sanghuangporus sp.Keywords: Antibacterial activity, Anticancer activity, Sanghuangporus sp.1, Mycelia

Crucial Role of Heme Oxygenase-1 on the Sensitivity of Cholangiocarcinoma Cells to Chemotherapeutic Agents

Cancer cells acquire drug resistance via various mechanisms including enhanced cellular cytoprotective and antioxidant activities. Heme oxygenase-1 (HO-1) is a key enzyme exerting potent cytoprotection, cell proliferation and drug resistance. We aimed to investigate roles of HO-1 in human cholangiocarcinoma (CCA) cells for cytoprotection against chemotherapeutic agents. KKU-100 and KKU-M214 CCA cell lines with high and low HO-1 expression levels, respectively, were used to evaluate the sensitivity to chemotherapeutic agents, gemcitabine (Gem) and doxorubicin. Inhibition of HO-1 by zinc protoporphyrin IX (ZnPP) sensitized both cell types to the cytotoxicity of chemotherapeutic agents. HO-1 gene silencing by siRNA validated the cytoprotective effect of HO-1 on CCA cells against Gem. Induction of HO-1 protein expression by stannous chloride enhanced the cytoprotection and suppression of apoptosis caused by anticancer agents. The sensitizing effect of ZnPP was associated with increased ROS formation and loss of mitochondrial transmembrane potential, while Gem alone did not show any effects. A ROS scavenger, Tempol, abolished the sensitizing effect of ZnPP on Gem. Combination of ZnPP and Gem enhanced the release of cytochrome c and increased p21 levels. The results show that HO-1 played a critical role in cytoprotection in CCA cells against chemotherapeutic agents. Targeted inhibition of HO-1 may be a strategy to overcome drug resistance in chemotherapy of bile duct cancer

Investigation of antibacterial and anti-cancer activities of Streptomyces sp SRF1 culture filtrate

Purpose: To evaluate the antibacterial activity and cytotoxic effects of Streptomyces sp. SRF1 culture filtrate extract against breast cancer cell line.Methods: The activity of the extract against Gram-positive and Gram-negative bacteria was initially screened by an agar-well diffusion method. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were measured by broth microdilution method. Time-kill assays were also performed, and extract-induced morphological and ultrastructural changes to bacterial cells were investigated. Sulforhodamine B (SRB) assay was performed to determine the cytotoxicity of the extract against the human breast cancer cell line, MCF-7.Results: Antibacterial activity by the extract was detected against four strains of Gram-positive pathogens including one strain of methicillin-susceptible Staphylococcus aureus (MSSA) and 3 strains of methicillin-resistant Staphylococcus aureus (MRSA) - with low MIC and MBC values. This activity was bactericidal after 6 h exposure. Morphological alterations were detected on the cell surface of both MSSA and MRSA. The extract also inhibited MCF-7 cell growth with half-maximal concentration (IC50) of 211.67 ± 33.95 μg/mL in 72 h.Conclusions: Streptomyces sp. SRF1 culture filtrate extract exhibits potent antibacterial and anticancer activities and thus, represents a potential source of antibacterial and anticancer drugs.Keywords: Antibacterial activity, Anti-breast cancer, Staphylococcus aureus, Streptomyces sp. SRF

Cytotoxic, colony formation and anti-migratory effects of Spilanthes acmella (Asteraceae) aerial extract on MCF-7 cells and its cream formulation

Purpose: To determine anti-breast cancer activities of Spilanthes acmella (S. acmella) extract.Methods: S. acmella was macerated with 95% ethanol. Phenolic, flavonoid content and antioxidant activity of the extract were assessed using Folin  Ciocalteu method, aluminum chloride (AlCl3) colorimetric method and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, respectively. Cytotoxicity, colony formation and cell migration suppression on MCF-7 cells, representing anti-breast cancer effects, were also evaluated by sulforhodamine B (SRB), clonogenic and wound healing assays, respectively. Creams containing the extract were formulated and then characterized in terms of their physical appearance, viscosity and pH, before and after stability testing.Results: The crude extract contained phenolic content of 62.8 ± 5.2 mg gallic acid equivalent/g and flavonoid content of 375.6 ± 20.1 mg rutin equivalent/g. The results showed that the extract exhibits antioxidant effect with half-maximal inhibitory concentration (IC50) of 1.2 ± 0.1 mg/mL. It showed cytotoxicity on MCF-7 cells with IC50 of 37.1 ± 1.1 μg/mL in 48 h and inhibited colony formation of cells with IC50 of 44.9 ± 1.3 μM. In addition, it demonstrated an anti-migration effect at a concentration of 50 μg/mL. The developed creams displayed good physical appearance and maintained stable physical properties overt a two-month period. Conclusion: S. acmella extract exhibits potential anti-breast cancer activity. The cream containing the extract is promising for the topical treatment of breast cancer. Keywords: Spilanthes acmella, Antioxidant, Anti-breast cancer, Flavonoids, Phenolics, Topical crea

Evaluation of antioxidant and anticancer effects of Piper betle L (Piperaceae) leaf extract on MCF-7 cells, and preparation of transdermal patches of the extract

Purpose: To determine the antioxidant and anticancer effects of Piper betle (P. betle) leaf extract on human breast cancer MCF-7 cells, and to develop transdermal patches containing the extract. Methods: The leaf extract of P. betle was prepared by maceration method, and its antioxidant activity was evaluated using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. Cytotoxicity and suppression of cell migration (indices of anticancer activity) were also assessed in MCF-7 cells by sulforhodamine B (SRB) and wound healing assays, respectively. Transdermal patches were developed using the casting method, and the resultant patches were evaluated with regard to their physical appearance and mechanical properties before and after a stability test. Results: The extract exhibited antioxidant activity with half-maximal inhibitory concentration (IC50) of 30.0 ± 0.1 µg/mL. It also showed cytotoxicity with an IC50 of 114.3 ± 14.9 µg/mL, and significantly suppressed the migration of MCF-7 cells at a dose of 25 µg/mL. Based on desirable characteristics, patch base formulations containing 4.2 % pectin, 0.4 % hydroxyl propyl methylcellulose (HPMC), 0.4 % polyvinyl pyrrolidine K-90 (PVP-K90) and 3 % propylene glycol (PG) were selected for incorporation into the extract. Conclusion: Leaf extract of P. betle exhibits potential anti-breast cancer properties. A transdermal patch containing 0.03 % of the extract can be successfully developed for treatment of breast cancer

Cytotoxic effects of Saccharomyces cerevisiae TC6 and Lactobacillus brevis TBRC 3003 isolated from Thai fermented foods

Purpose: To determine the cytotoxic effect, anti-colony formation effect and antimigratory effect of Saccharomyces cerevisiae TC6 isolated from Thai water kefir, and Lactobacillus brevis TBRC 3003 isolated from picked cabbage. Methods: Crude microbial extracts were obtained from whole cultures (cells and broths) using ethyl acetate as extracting solvent, and the dried extracts were redissolved in ethanol before use. Cytotoxic, antiproliferative and antimigratory effects of the two microbial extracts on MCF-7, HepG2, and HeLa were tested using 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetra zolium bromide (MTT), clonogenic formation and wound healing assays. Results: Lb. brevis TBRC 3003 showed the highest cytotoxicity toward HepG2 cells (IC50 of 669.72 µg/mL), while S. cerevisiae TC6 showed the highest cytotoxicity against MCF-7 (IC50 of 691.49 µg/mL) and HeLa (IC50 of 379.16 µg/mL) based on MTT assay. Anti-colony formation test showed that S. cerevisiae TC6 was most the most effective in inhibiting colony formation of HepG2 (IC50 of 311.12 µg/mL) and HeLa (IC50 of 494.64 µg/mL), while Lb. brevis TBRC 3003 was the most potent in inhibiting colony formation of MCF-7 (IC50 of 267.88 µg/mL). Conclusion: Both microbes can potentially be implemented in functional foods as bio-therapeutics with chemopreventive properties against breast, liver and cervical cancers