Antiproliferative and apoptotic effects of high-dose vitamin C in cholangiocarcinoma cell line

Purpose: To evaluate the antiproliferative and apoptotic effects of high-dose vitamin C incholangiocarcinoma cell line (CCA).Methods: Sulforhodamine B colorimetric assay was used to determine cell proliferation in human K100- CCA cell lines. The expression of anti-apoptotic Bcl-2 protein/ pro-apoptotic Bax protein was evaluated by Western blot analysis.Results: Ascorbic acid inhibited the growth of CCA in a dose-dependent manner. The half-maximal inhibitory concentration (IC50) of ascorbic acid on K100 at 24 h was 55 ± 9.4 mM. It was observed that treatment with ascorbic acid resulted in the reduction of glutathione and increased hydrogen peroxide contents in the cells. Moreover, disturbance of mitochondrial membrane potential occurred in ascorbic acid-treated cells in a dose-dependence manner. The ratio of Bcl2/Bax decreased in CCA treated with ascorbic acid.Conclusion: The results show that the anti-proliferation effect of ascorbic acid in CCA may be attributable to the modulation of expressions of Bcl-2 and Bax proteins and dissipation of the mitochondrial electrochemical potential gradient, which is an early event leading to apoptosis.Keywords: Vitamin C, Cholangiocarcinoma, Apoptosis, Glutathione, Mitochondrial membrane potentia

Protective Effects of Streblus asper

This study investigated the effects of Streblus asper leaf extract (SA) on reactive oxygen species (ROS) in SK-N-SH cell culture and on motor functions and behaviors in MPTP-treated C57BL/6 mice. SK-N-SH cell viability after incubation with SA for 24 h was measured by MTT assay. Intracellular ROS levels of SK-N-SH cells were quantified after pretreatment with SA (0, 200, 600, and 1000 µg/mL) in the presence of H2O2 (300 µM). Male C57BL/6 mice were force-fed with water or 200 mg/kg/day SA for 32 days. Intraperitoneal injection of MPTP was used to induce Parkinson’s disease-like symptoms. Catalepsy, beam balance ability, olfactory discrimination, social recognition, and spontaneous locomotor activity were assessed on days 19, 21, 23, 26, and 32, respectively. In cell culture, SA at 200, 600, and 1000 µg/mL significantly decreased ROS levels in H2O2-treated SK-N-SH cells. MPTP-treated C57BL/6 mice showed a significant change in all parameters tested when compared to the control group. Pretreatment and concurrent treatment with 200 mg/kg/day SA could antagonize the motor and cognitive function deficits induced by MPTP. The results show that SA possesses anti-Parkinson effects in MPTP-treated C57BL/6 mice and that reduction in ROS levels might be one of the mechanisms

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

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

Antitumor effects of candidone extracted from Derris indica (Lamk) Bennet in cholangiocarcinoma cells

Purpose: To investigate the antitumor effect of candidone extracted from Derris indica, against human cholangiocarcinoma (CCA) cells.Methods: Candidone was purified from the hexane extract of Derris indica fruit. CCA cell lines, KKUM156 and KKU-M213, were treated with candidone. Sulforhodamine B (SRB) assay and acridine orange/ethidium bromide (AO/EB) staining were used to investigate the effects of candidone on cell proliferation and induction of apoptosis, respectively. The effect on cell migration was assessed by wound healing assay. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was employed to assess the effects of candidone on the expression of genes that regulate proliferation and apoptosis.Results: Candidone exerted strong anticancer effects on CCA cells. The agent suppressed CCA cell proliferation and induced apoptotic cell death. RT-qPCR assay revealed that candidone significantly increased the expression of anti-proliferative and pro-apoptotic genes, including p21 and Bax, and decreased the expression of anti-apoptotic genes, including Bcl-2 and survivin. Moreover, candidone inhibited the migration of CCA cells induced by IGF-1.Conclusion: Candidone exhibits potent antitumor effect on CCA cells. These findings suggest that candidone is potentially suitable for the management of CCA and, therefore, warrants further investigation.Keywords: Candidone, Derris indica, Cholangiocarcinoma, Cytotoxicity, Apoptosi

Suppression of NAD(P)H-quinone oxidoreductase 1 enhanced the susceptibility of cholangiocarcinoma cells to chemotherapeutic agents

La lateralidad de la jugadora con el rol de base puede ser sólo una característica individual, o bien un factor influyente en el juego de su equipo, y por tanto, una herramienta a considerar por parte del equipo rival. El trabajo de investigación se centra en la necesidad de reflejar si la lateralidad de una jugadora con rol de base en baloncesto, al iniciar la acción de juego (bote o pase) influye de manera determinante en la toma de decisiones. El contexto de trabajo utilizado es la segunda categoría de baloncesto femenino nacional, Liga Femenina 2. En la observación han participado varios equipos de dicha categoría, con total de 10 partidos analizados y un total de 170 situaciones de juego analizadas (N=170). Para establecer las relaciones entre las variables se utilizó el Coeficiente de Contingencia. Los resultados muestran una relación estadísticamente significativa de la lateralidad de la base y el lado del campo en el que se empieza a jugar (X2 (1, N=170) = 4.789, p<.05)Laterality of the point-guard can be an individual characteristic, or in the other hand, an important factor in the offensive patterns of the team. In this way, it will be a useful information for the other team. This research has focused in the laterality of the point guard, and it influences in the decision making. The League studied was the second female Spanish basketball league, LF2. 10 matches were observed and 170 game situations analyzed (N=170). Crosstabs Command and the Contingency Coefficient were used to establish the relationship between variables. The results shows a significant relationship between the laterality of the point guard and the side of the field where the attack begin (X2 (1, N=170) = 4.789, p<.05)peerReviewe

Effect of lemongrass water extract supplementation on atherogenic index and antioxidant status in rats

Cymbopogon citratus (DC) Stapf., commonly known as lemongrass, possesses strong antioxidant and cardiotonic properties. Lemongrass water extract contains several polyphenolic compounds including gallic acid, isoquercetin, quercetin, rutin, catechin and tannic acid. Rutin, isoquercetin catechin and quercetin are the flavonoids most abundantly found in the extract. The extract significantly decreased total cholesterol, low-density lipoprotein and atherogenic index in rats after treatment (p < 0.05). Expression of genes and protein of sterol regulatory element binding protein-1c (SREBP1c) and HMG-CoA reductase (HMGR) was also lowered significantly in treated groups (p < 0.05). Moreover, serum antioxidant capacity increased in treated rats in comparison with untreated ones (p < 0.05) and was associated with decreased serum lipid peroxidation

Ferulic Acid Alleviates Changes in a Rat Model of Metabolic Syndrome Induced by High-Carbohydrate, High-Fat Diet

Agricultural Research Development Agency Fund, the Thailand Research Fund, the Khon Kaen University Research Fund and the Invitation Research Fund, Faculty of Medicine, Khon Kaen University. Senaphan K. was supported by a scholarship (PHD/0048/2553) from the Royal Golden Jubilee PhD Program, the Thailand Research Fun