Antioxidant, Apoptotic, and Wound Healing Effects of Xantolis cambodiana Extracts in Normal Human Dermal Fibroblasts.

Xantolis cambodiana has demonstrated significant antioxidant properties; however, the mechanisms underlying its protective effects against oxidative stress in cellular systems remain unexplored. This work investigated the efficacy of methanolic extracts in exhibiting oxidative damage and examined their mechanisms. The methanolic extract had a high phenolic content (116.89 ± 29.01 mg GAE/g FW) and exhibited scavenging of 2,2-diphenyl-1-picrylhydrazyl radicals with an IC50 value of 42.35 ± 9.20 µg/ml. In addition, it had the highest antioxidant activity based on ferric-reducing antioxidant power (467.45 ± 50.74 mg AA/100 g). Normal human dermal fibroblast (NHDF) cells were pretreated with the methanolic extract in a hydrogen peroxide (H2O2; 500 mM)-induced oxidative stress model, which resulted in a significant decrease in apoptosis and autophagy. Not only did the methanolic extract reduce mitochondrial membrane potential, it also stimulated NHDF cell migration and reduced reactive oxygen species production through mitochondrial dysfunction in the H2O-induced stress model. These findings suggested that the methanolic extract (25 µg/ml) attenuated H2O2-induced oxidative stress in NHDF cells, significantly reducing apoptosis, autophagy, and mitochondrial dysfunction. Thus, this extract has the potential to support the wound healing process due to its antioxidant activity.

Microwave Sensor for the Determination of DMSO Concentration in Water-DMSO Binary Mixture.

This research aims to develop a microwave sensor to accurately measure the concentration of dimethyl sulfoxide (DMSO) in water-DMSO binary mixtures. The proposed sensor will utilize microwave frequency measurements to determine the DMSO concentration, providing a non-invasive and efficient method for analyzing DMSO solutions. The research will involve the design, fabrication, and testing of the sensor, as well as the development of an appropriate calibration model. The outcomes of this study will contribute to improved monitoring and quality control in various fields, including pharmaceuticals, chemical synthesis, and biomedical research. The binary mixtures of dimethyl sulfoxide (DMSO) and water with varying concentrations were investigated in the frequency range of 1 GHz to 5 GHz at room temperature using a microwave sensor. The proposed microwave sensor design was based on an interdigital capacitor (IDC) microstrip antenna loaded with a hexagonal complementary ring resonator (HCRR). The performance of the sensor, fabricated using the print circuit board (PCB) technique, was validated through simulations and experiments. The reflection coefficient (S11) and resonance frequency (Fr) of binary mixtures of DMSO and water solutions were recorded and analyzed for DMSO concentrations ranging from 0% v/v to 75% v/v. Mathematical models were developed to analyze the data, and laboratory tests showed that the sensor can detect levels of DMSO/water binary mixtures. The sensor is capable of detecting DMSO concentrations ranging from 0% v/v to 75% v/v, with a maximum sensitivity of 0.138 dB/% for S11 and ΔS11 and 0.2 MHz/% for Fr and ΔFr at a concentration of 50% v/v. The developed microwave sensor can serve as an alternative for detecting DMSO concentrations in water using a simple and cost-effective technique. This method can effectively analyze a wide range of concentrations, including highly concentrated solutions, quickly and easily.

Reactogenicity and immunogenicity of the intradermal administration of BNT162b2 mRNA vaccine in healthy adults who were primed with an inactivated SARS-CoV-2 vaccine.

Because of the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), safe and effective vaccines are urgently required. The shortage of effective vaccines is a major challenge in many developing countries. We studied intradermal (ID) fractional dose BNT162b2 mRNA (Comirnaty®, Pfizer-BioNTech) as a booster dose in healthy adults who were previously immunized with an inactivated SARS-CoV-2 vaccine. This is a retrospective cohort study that included healthy adults who were immunized with two doses of inactivated SARS-CoV-2 vaccine and received a booster dose with ID fractional dose or intramuscular (IM) full-dose BNT162b2 mRNA between August 1 to August 15, 2021. The primary endpoint was safety that included local and systemic adverse reactions. The secondary endpoints were levels of SARS-CoV-2 spike protein receptor-binding domain IgG antibody (anti-S-RBD IgG) and neutralizing antibody activity against the Delta variant (B.1.617.2) using surrogate viral neutralization test (sVNT) 3 weeks after the booster dose. A total of 43 healthy adults (median age of 31 years) were included in the study; among them, 23 participants received ID fractional dose (6 µg) BNT162b2 mRNA, and 20 participants received IM full-dose (30 µg) BNT162b2 mRNA. No serious adverse reactions were observed. Local adverse reactions occurred more frequently in the ID group. No differences were observed in the baseline level of anti-S-RBD IgG (289 vs 286 AU/mL, p > 0.9, in the ID and IM groups, respectively). After booster, anti-S-RBD IgG titer increased to 13294 (9255-19573) AU/mL in the ID group and 23456 (16943-38539) AU/mL in the IM group. All participants in the IM group and 95.6 % of participants in the ID group had seroconversion evaluated by sVNT (≥68 % inhibition to the Delta variant). ID administration of BNT162b2 mRNA was safe and well-tolerated and generated a robust immune response. Therefore, ID delivery of the BNT162b2 mRNA vaccine has the potential for a dose-sparing strategy.

The seed extract of Oroxylum indicum suppresses cell proliferation, migration and promotes apoptosis in cervical cancer cells.

The herb Oroxylum indicum has been used for treating several diseases and also has anticancer activity. This research examined the anticancer effects of ethanolic extract of O. indicum seeds on HeLa cervical cancer cells and investigates underlying mechanisms. The data indicated that the extract inhibited HeLa cells growth with low IC50 values and arrested the cell cycle at G0/G1 phase at a dose of 50 µg/mL. Moreover, this extract produced a marked increase in the apoptosis of cancer cells which was demonstrated by acridine orange/ethidium bromide (AO/EB) staining and was significant at a dose of 250 µg/mL for 24 h. Correlating with apoptotic data by flow cytometric analysis, the results indicated that viable cells were significantly reduced and late apoptotic cells were induced starting at the dose of 50 µg/mL of the extract. These extracts significantly induced cancer cell apoptosis which was mediating through reduction of mitochondrial function by stimulating intracellular reactive oxygen species (ROS) formation. Furthermore, the extract caused inactivation of cervical cancer cell migration and was detected with the wound healing method. The data in this study strongly indicated that O. indicum seed extract has powerful activity against cervical cancer.

Effect of cold plasma and elicitors on bioactive contents, antioxidant activity and cytotoxicity of Thai rat-tailed radish microgreens.

BACKGROUND: Raphanus sativus var. caudatus or Thai rat-tailed radish (RTR) contains glucosinolates and isothiocyanates with chemopreventive effects; however, only mature plants have been investigated to date. Thus, the present study aimed to determine isothiocyanates, phenolic compounds and flavonoid compounds, antioxidant activity, cytotoxicity, and antiproliferative activity of RTR microgreens grown from seeds treated with cold plasma (21 kV for 5 min), organic elicitor (160 mmol L-1 NaCl, 10 mmol L-1 CaCl2 or 176 mmol L-1 sucrose) or both in combination. Seeds were germinated on vermiculite and sprayed with deionized water or elicitor for 7 days before harvest. RESULTS: Cold plasma had insignificant effect on growth, whereas NaCl and CaCl2 increased fresh weight. Plasma with CaCl2 led to the highest total isothiocyanate (ITC) content [1.99 g kg-1 dry weight (DW)] in RTR microgreens containing raphasatin as the only ITC detected. Plasma treatment gave the highest total phenolic content (7.56 mg gallic acid equivalents g-1 DW), antioxidant activity from a 2,2-diphenyl-1-picrylhydrazyl assay (7.70 mg trolox equivalents g-1 DW) and ferric reducing antioxidant power assay (21.72 mg Fe2+ g-1 DW). Microgreen extracts from plasma showed an IC50 value of 29.28 and 13.83 µg mL-1 towards MCF-7 and HepG2, respectively, with inhibitory properties on matrix metalloproteinase (MMP)-2 and MMP-9 proteins. Plasma enhanced Bax and Caspase-3 gene expression but reduced Bcl-2 and MMP-9 expression, indicating activation of apoptosis. CONCLUSION: Cold plasma shows promise as an innovative tool to enhance bioactive compounds with chemopreventive benefits in microgreens. © 2020 Society of Chemical Industry.

Inhibition of cell proliferation and migration by Oroxylum indicum extracts on breast cancer cells via Rac1 modulation.

In this study, we investigated how Oroxylum indicum leaf and fruit extracts affect the viability and migration of MCF-7 breast cancer cells and the mechanisms of action responsible for these effects. MCF-7 cells treated with the extracts were examined using the sulforhodamine B, colony formation and caspase 3 activity assays, and by Western blotting. O. indicum extracts were found to inhibit MCF-7 cell growth in a concentration- and time-dependent manner, with 48 h IC50 values of 57.02 ± 2.85 µg/mL and 131.3 ± 19.2 µg/mL for leaf and fruit extracts, respectively. Further, the O. indicum leaf extract caused a reduction in MCF-7 cell viability, induction of MCF-7 cell apoptosis and ROS formation, and an increase in caspase 3 activity. Also, the two extracts inhibited MCF-7 cell migration and reduced both MMP 9 and ICAMP1 gene expression and MMP9 protein expression. Additionally, O. indicum extracts greatly reduced expression of the cell cycle regulatory protein Rac1 in the mevalonate pathway. In summary, O. indicum leaf and fruit extracts reduce breast cancer cell growth, cell viability and cell migration. O. indicum constituents could, therefore, be useful for augmenting the activity of chemotherapeutic drugs employed to treat breast cancer.

Comparative Study on the Effect of Aqueous and Ethanolic Mycelial Extracts from Polycephalomyces nipponicus (Ascomycetes) against Human Breast Cancer MCF-7 Cells.

Recently, the entomopathogenic fungus Polycephalomyces nipponicus has been determined to be a prolific producer of bioactive compounds that have both antibacterial and antimarial activities, but the anticancer effects of the mycelial extracts have not been well studied. The present study investigates the effects and mechanisms of action of P. nipponicus extracts that are responsible for cell death in the human breast cancer MCF-7 cell line. The results showed that the 50% ethanol extract had greater anticancer activity than the aqueous extract. The 50% ethanolic extract inhibited cell growth at concentrations of just 109.75 ± 3.54 µg mL-1 for 72 h. The aqueous extract's activity was > 400 µg mL-1 against MCF-7 cells. Similar results were obtained from the colony formation assay. Moreover, the 50% ethanolic extract caused a significant increase in the distribution of cells at the G1 phase in a dose-dependent manner. The aqueous extract induced MCF-7 cells to arrest at the G2/M phase in a dose-dependent manner. These were supported by the reduction in the cyclin-dependent kinase (cdk)2, cdk4, and cdk6 genes' expression levels along with an induction of the cyclin-dependent kinase inhibitor p21 in MCF-7 cells after treatment with the extracts. In conclusion, P. nipponicus may be useful for breast cancer prevention and treatment in the future.

Inhibition of growth and migration of cholangiocarcinoma cells by pamidronate.

Pamidronate has been hypothesized to effectively inhibit cancer cell growth and metastasis in bone tissue. Furthermore, pamidronate (Pami) exerts various direct effects against several cancer cell types, including growth and migration. The present study aimed to determine the underlying mechanism of Pami's effect on the proliferation and migration of cholangiocarcinoma (CCA) cells. KKU-100 cells were used to determine the effects of Pami on cell death and migration. The following were assessed: Sulforhodamine B, colony formation, apoptosis via flow cytometry, reactive oxygen species (ROS) production and caspase-3 activity. In addition, the effects of the test compound on the mevalonate (MVA) signaling pathway were determined via western blotting and reverse transcription-quantitative PCR. Cell migration was observed via wound healing, Matrigel and gelatin zymography. The results indicated that Pami induced CCA cell death and inhibited colony formation in a dose-dependent manner, with IC50 values of 444.67±44.05 µM at 24 h and 147.33±17.01 µM at 48 h. Furthermore, Pami treatment suppressed colony formation at a lower concentration than growth inhibition with IC50 values of 5.36±0.31 µM. The mechanism of growth inhibition was determined to potentially be associated with increased ROS generation and stimulated caspase-3 enzyme activity, leading to the induction of apoptosis. Furthermore, Pami treatment interfered with the MVA signaling pathway by reducing Rac1 protein levels and modulating the gene and protein expression of RhoA. Furthermore, Pami suppressed CCA cell migration by decreasing matrix metalloproteinase (MMP)2 and MMP9 levels. Additionally, Pami treatment activated CCA cell death and inhibited CCA migration at low concentrations. Pami significantly decreased the protein expression levels of Rac1 in the MVA signaling pathway and may therefore be beneficial for developing a novel chemotherapeutic method for CCA.

Antiproliferative and antimigratory activities of bisphosphonates in human breast cancer cell line MCF-7.

Bisphosphonates (BPs) are antiresorptive drugs that act as effective inhibitors of cancer cell proliferation. However, not all bisphosphonates are equally effective against breast cancer cells in vitro. The present study investigated the extent to which three BPs decrease the viability of MCF-7 human breast cancer cells, stimulate cell apoptosis and inhibit cell migration by modulating proteins in the mevalonate pathway. The three BPs exerted direct anticancer effects against MCF-7 cells in a dose- and time-dependent manner, with pamidronate demonstrating the highest efficacy. In addition, the BPs inhibited colony formation ability. The activity of BPs against MCF-7 cells was inhibited by the mevalonate product geranylgeranyl pyrophosphate, which was potentiated by doxorubicin. It was also identified that BPs modulated Ras-related C3 botulinum toxin substrate 1, Ras homolog gene family member A and cell division control protein 42 homolog gene expression. Consistent with the observed growth inhibitory effects, BPs also inhibited the cell cycle by promoting G1 phase arrest and the downregulation of cyclin D1 and upregulation of p21. Additionally, BPs were revealed to induce reactive oxygen species expression, caspase-3 activity and increase the mitochondrial transmembrane potential, which was associated with apoptosis. BP-induced cancer cell apoptosis was detected by acridine orange/ethidium bromide staining and flow cytometry analysis, and was identified to be associated with the induction of caspase-3 and cytochrome c protein expression. Furthermore, BPs significantly decreased cancer cell migration in a dose-dependent manner and reduced matrix metallopeptidase-9 protein expression. In summary, the current study demonstrated that BPs exhibited a direct anticancer effect and an antimigratory effect on MCF-7 cells. These findings suggest that BPs may be developed as a therapeutic option for breast cancer and may serve as sensitizing chemotherapeutic agents.

Oroxylum indicum (L.) extract protects human neuroblastoma SH­SY5Y cells against ß­amyloid­induced cell injury.

It has been reported that amyloid ß peptide, the major component of senile plaques, serves a critical role in the development and progression of Alzheimer's disease (AD) by generating reactive oxygen species (ROS), leading to oxidative stress. The aim of the present study was to investigate the protective effect of Oroxylum indicum (L.) extract against Aß25­35­induced oxidative stress and cell injury using SH­SY5Y cells as a model, and at exploring the underlying mechanisms. The results revealed that the exposure of cells to 20 µM Aß25­35 significantly increased cellular oxidative stress, as evidenced by the increased ROS levels. Aß25­35 treatment also increased caspase­3/7 activity and lactate dehydrogenase (LDH) release, and caused viability loss. Oroxylum indicum treatment not only attenuated the generation of ROS and suppressed caspase­3/7 activity but also reduced the neurotoxicity of Aß25­35 in a concentration­dependent manner, as evidenced by the increased cell viability and decreased LDH release. Treatment with Oroxylum indicum also increased superoxide dismutase (SOD) and catalase (CAT) activity, increased the phosphorylation of Akt and cAMP­responsive element binding protein (CREB), and contributed to the upregulation of Bcl­2 protein. In combination, these results indicated that Oroxylum indicum extract could protect SH­SY5Y cells against Aß25­35­induced cell injury, at least partly, by inhibiting oxidative stress, increasing SOD and CAT activity, attenuating caspase 3/7 activity and promoting the cell survival pathway, Akt/CREB/Bcl­2. The approach used in the present study may also be useful for preventing the neurotoxicity induced by Aß in AD and related neurodegenerative diseases. Further studies investigating the activity of Oroxylum indicum extract in vivo are now required.

Identification of ortho-hydroxy anilide as a novel scaffold for lysine demethylase 5 inhibitors.

Fe(II)/α-ketoglutarate-dependent lysine demethylases (KDMs) are attractive drug targets for several diseases including cancer. In this study, we designed and screened ortho-substituted anilides that are expected to function as Fe(II) chelators, and identified ortho-hydroxy anilide as a novel scaffold for KDM5A inhibitors. Treatment of human lung cancer A549 cells with a prodrug form of 4-carboxy-2-hydroxy-formanilide (9c) increased trimethylated lysine 4 on histone H3 level, suggesting KDM5 inhibition in the cells.

Evaluation of Antibacterial and Anticancer Activities of the Medicinal Fungus Ophiocordyceps sobolifera (Ascomycetes) from Thailand.

Eleven strains of an entomopathogenic fungus, isolated and identified as Ophiocordyceps sobolifera, were screened for activity against 5 strains of Gram-negative and 5 strains of Gram-positive bacteria. Four of the isolates, Cod-KK1634, Cod-KK1643, Cod-KS1601, and Cod-SN1626, had activity against the test strains of Grampositive bacteria. Of these 4 extracts, the Cod-KK1643 extract had the lowest minimum inhibitory and bactericidal concentrations. The Cod-KK1643 extract exhibited both concentration- and time-dependent bactericidal activity. Moreover, the Cod-KK1643 extract induced morphological alterations in bacterial cells, including decreased cell size, a crushed appearance, and cell lysis. It is surprising to note that the extracts also inhibited MCF-7 cell proliferation, with a half-maximal inhibitory concentration of 47.09 ± 33.64 mg/mL after 72 hours. The extracts also inhibited MCF-7 cell migration, with the lowest percentage of relative closure achieved with Cod-KK1643. These findings represent what is to our knowledge the first information on the activities of O. sobolifera mycelial extracts against bacteria and the human breast cancer MCF-7 cell line.

Simvastatin potentiates doxorubicin activity against MCF-7 breast cancer cells.

Simvastatin is a low density lipoprotein-lowering drug that is widely used to prevent and treat cardiovascular disease by inhibiting the mevalonate pathway. Simvastatin also exhibits inhibitory effects on a number of types of cancer. In the present study, the effects of simvastatin on the activity of doxorubicin in the breast cancer MCF-7 cell line, and the mechanisms by which this interaction occurs were investigated. The effect of simvastatin and doxorubicin treatment, alone and in combination, on the growth of MCF-7 cells was evaluated by a sulforhodamine B and colony formation assay. To delineate the mechanisms of cell death, the following parameters were measured: Reactive oxygen species (ROS) production using the fluorescence probe dihydroethidium; caspase 3 activity by the fluorometry method; gene expression by quantitative polymerase chain reaction; and apoptotic- and proliferative-related protein levels by western blotting. MCF-7 cell proliferation was significantly suppressed by 24-48 h treatment with simvastatin alone. Doses of 10-50 µM simvastatin also enhanced the cytotoxicity of doxorubicin against MCF-7 cells in a dose-dependent manner, and decreased the colony-forming ability of MCF-7 cells. Simvastatin alone or in combination with doxorubicin significantly increased ROS levels. Combination treatment significantly decreased expression of the cell cycle regulatory protein Ras-related C3 botulinum toxin substrate 1 and numerous downstream proteins including cyclin-dependent kinase (Cdk) 2, Cdk4 and Cdk6. Additionally, simvastatin in combination with doxorubicin significantly induced expression of the cyclin-dependent kinase inhibitor p21, increased cytochrome c and caspase 3 expression and reduced cyclin D1 expression. In conclusion, simvastatin acts synergistically with the anticancer drug doxorubicin against MCF-7 cells, possibly through a downregulation of the cell cycle or induction of apoptosis. Although additional studies are required, simvastatin and doxorubicin combination may be a reasonable regimen for the treatment of breast cancer.

Cytotoxic and antimigratory effects of Cratoxy formosum extract against HepG2 liver cancer cells.

The aim of the present study was to investigate the molecular mechanisms underlying Cratoxylum formosum (CF) Dyer-induced cancer cell death and antimigratory effects in HepG2 liver cancer cells. The cytotoxic, antiproliferative and antimigratory effects of CF leaf extract on human liver cancer HepG2 cell lines were evaluated using sulforhodamine B, colony formation, and wound healing assays. In addition, apoptosis induction mechanisms were investigated via reactive oxygen species (ROS) formation, caspase 3 activities, and mitochondrial membrane potential (ΔΨm) disruption. Gene expression and apoptosis-associated protein levels were measured by reverse transcription-quantitative polymerase chain reaction and western blotting. CF induced HepG2 cell death in a time- and dose-dependent manner with half maximal inhibitory concentration values of 219.03±9.96 and 124.90±6.86 µg/ml at 24 and 48 h, respectively. Treatment with CF caused a significant and dose-dependent decrease in colony forming ability and cell migration. Furthermore, the present study demonstrated that CF induced ROS formation, increased caspase 3 activities, decreased the ΔΨm, and caused HepG2 apoptosis. CF marginally decreased the expression level of the cell cycle regulatory protein, ras-related C3 botulinum toxin substrate 1 (rho family, small GTP binding protein Rac1) and the downstream protein, cyclin dependent kinase 6. Additionally, CF significantly enhanced p21 levels, reduced cyclin D1 protein levels and triggered cancer cell death. CF leaf extracts induced cell death, stimulated apoptosis and inhibited migration in HepG2 cells. Thus, CF may be useful for developing an anticancer drug candidate for the treatment of liver cancer.

Cratoxy formosum leaf extract inhibits proliferation and migration of human breast cancer MCF-7 cells.

In this study we investigated how Cratoxy formosum (CF) leaf extract affects the viability and migration of human breast cancer cells including the mechanism(s) responsible. Our results showed that CF leaf extract strongly induced MCF-7 cell death in a concentration- and time-dependent manner, with IC50 values of 85.70±4.52µg/mL and 53.74±3.02µg/mL at 24h and 48h, respectively. Additionally, CF leaf extract potentiated the activity of 4 anticancer drugs with the greatest synergy occurring between CF and 5-FU. CF leaf extract also caused a dose-dependent decrease in colony forming ability with IC50 values of 36.37+1.80 µg/mL and cell migration, with IC50 values of 43.68±0.86µg/mL. Moreover, CF significantly induced ROS formation, increased caspase 3 activities, and reduced the mitochondrial membrane potential, leading to cancer cell apoptosis and cell death. In addition, the extract inhibited cancer cell migration at 25µg/mL by reducing MMP 2 and MMP 9 protein expression. Moreover, CF leaf extracts strongly decreased expression of the cell cycle regulatory protein Rac1 and downstream protein, cdk6. CF leaf extract significantly stimulated p21 and this correlated with a reduction in cyclin D1 protein levels. In summary, CF leaf extract can inhibit cell proliferation, induce cell apoptosis, and reduce cell migration in the MCF-7 cell line. It could also be beneficial for enhancing the activity of anticancer drugs used to treat breast cancer.

Effects of Ethyl Acetate Extracts from the Polycephalomyces nipponicus Isolate Cod-MK1201 (Ascomycetes) against Human Pathogenic Bacteria and a Breast Cancer Cell Line.

This study aimed to identify a suitable organic solvent for extracting bioactive compounds from Polycephalomyces nipponicus and to evaluate the antibacterial and anticancer activities of the extracts obtained. Only extracts obtained with ethyl acetate exhibited antibacterial activity, so ethyl acetate was chosen for large-scale extraction. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Gram-positive and Gram-negative human pathogenic bacteria of the 3 ethyl acetate-derived extracts-ethyl acetate extract from P. nipponicus (PN-ME), ethyl acetate extract after defatting (PN-ME*), and ethyl acetate extract after refluxation (PN-ME')-were determined. PN-ME' exhibited the most potent activity, inhibiting 12 of the 18 test bacteria, especially Bacillus cereus ATCC 11778 and Vibrio cholera (O1) DMST 9700, with low MIC and MBC values. PN-ME* showed greater inhibitory activity than PN-ME. The effects of the extracts on bacterial cell morphology were also determined. After 120 minutes of treatment with PN-ME* or PN-ME', B. cereus ATCC 11778 exhibited an abnormal rod-shaped cell structure, with some cells elongated to multiple times their original size and others appearing collapsed. V cholera (O1) DMST 9700 cells showed shrinkage and the formed subsurface cavities. PN-ME* and PN-ME' also inhibited the growth of MCF-7 breast cancer cells. In conclusion, the fungal isolate P. nipponicus Cod-MK1201 represents a source of antibacterial and anti-breast cancer compounds.

Simvastatin and atorvastatin as inhibitors of proliferation and inducers of apoptosis in human cholangiocarcinoma cells.

AIMS: In this study, we investigated whether statins induce human cholangiocarcinoma (CCA) cell death and apoptosis, and examined the mechanism by which statins act on cells. MAIN METHODS: Four CCA cell lines, KKU-100, KKU-M055, KKU-M214, and KKU-M156 CCA cell lines were examined for HMGCR mRNA expression by the RT-PCR method. Two CCA cell lines, with low and high HMGCR mRNA expression, were used to evaluate the sensitivity to two statins, simvastatin and atorvastatin. Cytotoxic activity, antiproliferative activity, and cell migratory effects of the statins on CCA cells were evaluated using sulforhodamine B (SRB) and acridine orange/ethidium bromide (AO/EB), the colony formation assay, and wound healing assay, respectively. ROS formation was measured and apoptosis-related proteins were analyzed by Western blotting. KEY FINDINGS: Both statins induced KKU-100 and KKU-M214 cell death in a time- and dose-dependent manner. Statins induced cell death more potently in the KKU-100 cells exhibiting low HMGCR expression than the KKU-M214 cells which had high HMGCR expression. Simvastatin was more potently cytotoxic than atorvastatin with lower IC50 values. Treatment with statins also caused a concentration-dependent decline in colony forming ability and cell migration. Both statins induced reactive oxygen species (ROS) formation in KKU-100 cells, but not in KKU-M214 cells. Simvastatin enhanced the release of cytochrome c, caspase 3, and increased p21 levels, especially for the KKU-100 cells. SIGNIFICANCE: Statins induced CCA cell death, inhibited cell migration, and induced apoptosis. Cell death was probably induced via the mitochondrial pathway. Statins could potentially be developed as novel chemotherapeutic agents for CCA.

Cytoprotective effects of ferritin on doxorubicin-induced breast cancer cell death.

Ferritin is a major iron storage protein and essential for iron homeostasis. It has a wide range of functions in the body including iron delivery, immunosuppression, angiogenesis, and cell proliferation. Ferritin is overexpressed in many cancer cells, but its precise role in cancer is unclear. In the present study, we examined the functional roles of ferritin in protecting the MCF-7 breast cancer cell line against treatment with the chemotherapeutic agent doxorubicin. The effects of ferritin (human liver ferritin) and doxorubicin on the human MCF-7 breast cancer cell line were evaluated using the cell viability assay. The impact of decreasing ferritin light chain (FTL) and ferritin heavy chain (FTH) expression on doxorubicin sensitivity was assessed using siRNA. Reactive oxygen species (ROS) was also measured using the fluorescence probe CM-H2DCFDA. The mechanism of modulated chemosensitivity was evaluated by western blot analysis. Ferritin treatment activated MCF-7 cell proliferation in a concentration- and time-dependent manner. While treatment with doxorubicin alone significantly increased intracelullar ROS production, the addition of ferritin decreased this ROS formation, thereby reducing doxorubicin­induced MCF-7 cell death. The inhibition of FTL and FTH by siRNA sensitized cells to doxorubicin. Treatment with doxorubicin alone significantly induced the cell cycle­dependent kinase inhibitor protein p21, whereas ferritin reduced p21 expression. Thus, ferritin plays a critical role in protecting MCF-7 cells against the chemotherapeutic drug doxorubicin. A targeted reduction of ferritin may be a useful strategy for overcoming chemoresistance in breast cancer.

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.

NQO1 expression correlates with cholangiocarcinoma prognosis.

Cholangiocarcinoma (CCA) is a rare type of liver cancer with a very poor prognosis. The prevalence of CCA is markedly variable with the highest incidence in the northeast Thailand, followed by other parts of Southeast Asia and China. Currently, there is still no reliable biomarker for diagnosis or treatment. NADPH-quinone oxidoreductase 1 (NQO1) is a xenobiotic metabolizing enzyme detoxifying chemical stressors and antioxidants, thereby providing cytoprotection in normal tissues. However, NQO1 is over-expressed in some cancers, suggesting roles in carcinogenesis and tumor progression. In this study, we examined NQO1 activity in surgical specimens from CCA patients and found much higher values than in the adjacent normal tissues. Immunohistochemical analysis revealed strong staining in tumor epithelial elements, whereas the non-tumor bile ducts and liver parenchyma were weakly stained. NQO1 mRNA expression in tumor tissues was widely varied among 43 patients. A significant association was observed between high level of NQO1 expression and short overall survival time by the Cox proportional hazard ratio of 2.40, p<0.05. By histological classification, non-papillary adenocarcinoma was an independent predictor for poor prognosis with the hazard ratio of 2.79, p<0.05. NQO1 expression may serve as a prognostic biomarker for the CCA.