• Journal of Nutrition and Health
• /
• v.35 no.2
• /
• pp.192-200
• /
• 2002

Conjugated linoleic acid (CLA) is a collective term for positional and geometric isomers of octadecadienoic acid in which the double bonds are conjugated. CLA has anticancer activity in a variety of animal cancer models, and cis-9, trans-11 (c9t11) and trans-10, cis-12(t10c12) CLA are the most predominant isomers present in the synthetic preparations utilized in these animal studies. To compare the ability of c9t11, t10c12 and an isomeric mixture of CLA to inhibit TSU-Prl cell growth, cells were incubated in a serum-free medium with various concentrations of these fatty acids. The isomeric mixture inhibited cell growth in a dose-dependent manner (1-3 $\mu$M) with a 41 $\pm$ 1% inhibition observed at 3 $\mu$M concentration after 48 hours. T10c12 also inhibited cell proliferation in a dote-dependent manner, However, the efficacy and potency of this isomer was much greater than that of the isomeric mixture with a 49 $\pm$ 2% inhibition observed at 0.3 $\mu$M concentration after 48 hours. By contrast, c9t11 slightly increased cell proliferation. To determine whether the growth-inhibiting effect of CLA is related to the changes in production of insulin-like growth factors (IGF) and IGF-binding proteins (IGFBP) by these cells, serum-free conditioned media were collected. Immunoblot analysis of conditioned media using a monoclonal anti-IGF-II antibody showed that both the isomeric mixture and t10c12 inhibited secretion of both mature 7,500 Mr and higher Mr forms of pro IGF-II, whereas c9t11 had no effect. Ligand blot analysis with 125I-IGF-II revealed the presence of two types of IGFBPs : 24,000 Mr IGFBP-4 and 30,000 Mr IGFBP-6. The production of IGFBP-4 slightly decreased at the highest concentrations of the isomeric mixture and t10c12. These results indicate that CLA inhibits human prostate cancer cell growth, an effect largely due to the action of t10c12. The growth inhibition may result, at least in part, from decreased production of IGF-II and IGFBP-4 by these cells.

• Korean Journal of Plant Resources
• /
• v.24 no.2
• /
• pp.130-138
• /
• 2011

In this study, we evaluated the protective effects of the hot water extract from red bean (Phaseolus angularis) against oxidative DNA and cell damage induced by hydroxyl radical. The antioxidant activities were evaluated by hydroxyl radical and hydrogen peroxide scavenging assay, and $Fe^{2+}$-chelating assay. Although the extract with hot water didn't scavenge the hydroxyl radical, it removed and chelated hydrogen peroxide and ferrous iron necessary for the induction of hydroxyl radical by 71% and 64% at 200 ${\mu}g/ml$, respectively. Its protective effect on oxidative DNA damage was carried using ${\Psi}$X-174 RF I plasmid DNA comparing the conversion level of supercoiled form of the plasmid DNA into open-circular form and linear form and the expression level of phospho-H2AX in NIH 3T3 cells. In ${\Psi}$X-174 RF I plasmid DNA cleavage assay, it inhibited oxidative DNA damage by 96% at 200 ${\mu}g/ml$. Also, it decreased the expression of phospho-H2AX by 50.1% at 200 ${\mu}g/ml$. Its protective effect against oxidative cell damage was measured by MTT assay and the expression level of p21 protein in NIH 3T3 cells. In MTT assay for the protective effect against the oxidative cell damage, it inhibited the oxidative cell death and the abnormal cell growth induced by hydroxyl radical. Also, it inhibited p21 protein expression by 98% at 200 ${\mu}g/ml$. In conclusion, the results of the present studies indicate that hot water extract from red bean exhibits antioxidant properties and inhibit oxidative DNA damage and the cell death caused by hydroxyl radical.

• BMB Reports
• /
• v.53 no.2
• /
• pp.82-87
• /
• 2020

Circular RNAs (circRNAs), one kind of non-coding RNA, have been reported as critical regulators for modulating gene expression in cancer. In this study, microarray analysis was used to screen circRNA expression profiles of bladder cancer (BC) 5637 cells, T24 cells and normal control SV-HUC-1 cells. The data from the microarray showed that hsa_circ_0075828 (named circCASC15) was most highly expressed in 5637 and T24 cells. circCASC15 was highly expressed in BC tissues and cells. Overexpression of circCASC15 was closely associated with BC tumor stage and promoted cell proliferation significantly in vitro and in vivo. Mechanistically, circCASC15 could act as miR-1224-5p sponge to activate the expression of CREB1 to promote cell proliferation in BC. In short, circCASC15 promotes cell proliferation in BC, which might be a new molecular target for BC diagnosis and therapy.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.3
• /
• pp.1721-1724
• /
• 2013

Objective: To investigate the effect of epirubicin on soluble CD25 (sCD25) secretion by CD4+CD25+ regulatory T (Treg) cells isolated from diffuse large B-cell lymphoma (DLBCL) patients. Methods: Treg cells were isolated from the peripheral blood mononuclear cells isolated from the newly diagnosed DBLCL patients. The concentration of sCD25 in the supernatant was determined with a commercial sCD25 (IL-2R) enzyme-linked immunosorbent assay (ELISA) kit. The fluorescence intensity of CD25 was detected by flow cytometry. Results: Cell survival rate was significantly decreased along with the increase of epirubicin concentration after treatment for 24 h. There was also a significant difference in the concentration of sCD25 between the epirubicin group and the control group (P<0.01). A positive correlation between the Treg cells survival rate and the concentration of sCD25 was detected (r=0.993, P<0.01). When equal numbers of CD4+CD25+ Treg cells of the epirubicin group and the control group were cultured for another 24 h without epirubicin the CD25 fluorescence intensity on the surface of Treg cells was obviously higher in the epirubicin group than that in the control group (P<0.01), while the sCD25 concentration in the supernatant in the epirubicin group was significantly lower than that in the control group (P<0.05). Conclusion: Epirubicin may improve the body's immune functions by inhibiting the sCD25 secretion by Treg cells in DLBCL patients.

• Molecules and Cells
• /
• v.41 no.3
• /
• pp.188-197
• /
• 2018

Benzidine, a known carcinogen, is closely associated with the development of bladder cancer (BC). Epithelial-mesenchymal transition (EMT) is a critical pathophysiological process in BC progression. The underlying molecular mechanisms of mitogen-activated protein kinase (MAPK) pathway, especially extracellular regulated protein kinases 5 (ERK5), in regulating benzidine-induced EMT remains unclarified. Hence, two human bladder cell lines, T24 and EJ, were utilized in our study. Briefly, cell migration was assessed by wound healing assay, and cell invasion was determined by Transwell assay. Quantitative PCR and western blot were utilized to determine both gene expressions as well as protein levels of EMT and MAPK, respectively. Small interfering RNA (siRNA) was transfected to further determine ERK5 function. As a result, the migration and invasion abilities were enhanced, epithelial marker expression was decreased while mesenchymal marker expression was increased in human BC cell lines. Meanwhile, benzidine administration led to activation of ERK5 and activator protein 1 (AP-1) proteins, without effective stimulation of the Jun N-terminal kinase (JNK) or p38 pathways. Moreover, Benzidine-induced EMT and ERK5 activation were completely suppressed by XMD8-92 and siRNAs specific to ERK5. Of note, ERK1/2 was activated in benzidine-treated T24 cells, while benzidine-induced EMT could not be reversed by U0126, an ERK1/2 inhibitor, as indicated by further study. Collectively, our findings revealed that ERK5-mediated EMT was critically involved in benzidine-correlated BC progression, indicating the therapeutic significance of ERK5 in benzidine-related BC.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.5
• /
• pp.2253-2255
• /
• 2012

Objective: The objective of our present study was to assess the role of serum amyloid A (SAA) in stages and prognosis of renal cell carcinoma. Material and Methods: It was a hospital based retrospective study carried out in the Department of Medicine and Biochemistry of Manipal Teaching Hospital, Pokhara, Nepal between $1^{st}$ January 2008 and $31^{st}$ December 2011. The variables collected were SAA, CRP. Approval for the study was obtained from the institutional research ethical committee. Quantitative analysis of human SAA and C-reactive protein (CRP) was performed by radial immune diffusion (RID) assay for all cases. Results: Of the 422 total cases of renal cell carcinoma, 218 patients had normal and 204 abnormal SAA. SAA levels were grossly elevated in T3 stage ($122.3{\pm}SD35.7$) when compared to the mean for the T2 stage ($84.2{\pm}SD24.4$) (p value: 0.0001). Similarly, SAA levels were grossly elevated in M1 stage ($190.0{\pm}SD12.7$) when compared to the M0 stage ($160.9{\pm}SD24.8$) (p: 0.0001). There was no significant association with elevated CRP levels ($209.1{\pm}SD22.7$, normal $199.0{\pm}SD19.5$). Conclusion: The validity of SAA in serum as being of independent prognostic significance in RCC was demonstrated with higher levels in advanced stage disease.

• Korean Journal of Acupuncture
• /
• v.28 no.1
• /
• pp.61-69
• /
• 2011

Objectives : The purpose of this study is to investigate effects of White Ginseng-Ejung-tang acupuncture solution (EJ) on nitric oxide (NO) and of hydrogen peroxide production in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). Methods : Cell viability was measured by modified MTT assay. NO production was measured by Griess reagent assay. Hydrogen peroxide production was measured by dihydrorhodamine 123 (DHR) assay. Significant differences were examined by using a Student's t-test. Results : The results of the experiment are as follows. 1. EJ did not show cell toxicity against RAW 264.7 cells for 24 hr incubation at the concentrations of up to $200\;{\mu}g$/mL in RAW 264.7 cells. 2. EJ significantly inhibited NO production for 24 hr incubation in RAW 264.7 cells (p <0.05). 3. EJ significantly inhibited the LPS-induced production of NO for 24 hr incubation in RAW 264.7 cells (p <0.05). 4. EJ significantly inhibited the LPS-induced production of hydrogen peroxide for 16, 24, 40, 48, 64, and 72 hr incubation in RAW 264.7 cells (p <0.05). Conclusions : These results suggest that EJ has an anti-inflammtory property related with its inhibition of NO and hydrogen peroxide production in LPS-induced macrophages.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.5
• /
• pp.2015-2020
• /
• 2012

Oleanolic acid (OA) is a naturally occurring triterpenoid in food materials and is a component of the leaves and roots of Olea europaea, Viscum album L., Aralia chinensis L. and more than 120 other plant species. There are several reports validating its antitumor activity against different cancer cells apart from its hepatoprotective activity. However, antitumor activity against skin cancer has not beed studied well thus far. Hence the present study of effects of OA against HaCaT (immortalized keratinocyte) cells - a cell-based epithelial model system for toxicity/ethnopharmacology-based studies - was conducted. Radical scavenging activity ($DPPH{\cdot}$) and FRAP were determined spectrophotometrically. Proliferation was assessed by XTT assay at 24, 48 and 72 hrs with exposure to various concentrations (12.5-200 ${\mu}M$) of OA. Apoptotic induction potential of OA was demonstrated using a cellular DNA fragmentation ELISA method. Morphological studies were also carried out to elucidate its antitumor potential. The results revealed that OA induces apoptosis by altering cellular morphology as well as DNA integrity in HaCaT cells in a dose-dependent manner, with comparatively low cytotoxicity. The moderate toxicity observed in HaCaT cells, with induction of apoptosis, possibly suggests greater involvement of programmed-cell death-mediated mechanisms. We conclude that OA has relatively low toxicity and has the potential to induce apoptosis in HaCaT cells and hence provides a substantial and sound scientific basis for further validation studies.

• The Korea Journal of Herbology
• /
• v.38 no.1
• /
• pp.21-30
• /
• 2023

Objective : Gardeniae Fructus (GF) has bitter and cold nature. Thus, it has been traditionally prescribed in processed form roasted with ginger juice for patients with a weak stomach. This study investigated the effects of processed GF in tert-butyl hydroperoxide (tBHP)-treated gastric epithelial cells. Methods : Processed GF was made by applying 40% ginger juice or 10% ethanol for 24 h and then roasting at 150℃ for 5 minutes. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Mitochondrial membrane potential (MMP) was monitored by flow cytometry using the membrane permeable fluorescent dye Rh123. Protein expression was measured by Western blot analysis. Results : Cell viability was reduced by tBHP and restored by ethanol extract of GF (GFE). In the TUNEL assay, it was found that cell death by tBHP was due to apoptosis, and GFE had an anti-apoptotic effect. Processed GF roasted with ginger juice showed the best anti-apoptotic effect. Processed GF also inhibited MMP loss and restored tBHP-induced changes in expression levels of apoptosis-related proteins. Increased ROS production and GSH depletion after tBHP treatment were significantly reduced by processed GF. In addition, tBHP-induced activation of mitogen-activated protein kinase (MAPK) was inhibited by processed GF. Conclusion : These results demonstrate that the processed GF is able to protect gastric epithelial cells from oxidative stress-induced cell death with antiapoptotic and antioxidant activity. In addition, it shows that the processing of GF, which have been traditionally used for gastrointestinal protection, partially have scientific validity.

• The Korean Journal of Pesticide Science
• /
• v.6 no.2
• /
• pp.96-104
• /
• 2002

This study was carried out to investigate cytotoxicity of paraquat on NIH 3T3 fibroblasts, toxicity of paraquat and compensatory effects of 3-methylcholanthrene (3-MC) on the rat lung. In order to conduct MIT [3-(4,5-Dimethylthiazol-2-yl) -2,5-diphenyl -2H-tetrazolium-bromide] and NR (Neutral red) assay, the $5.0{\times}10^4cell/ml$ of NIH 3T3 fibroblast in each well of 24 multi-dish were cultured. After 24 hours, the cells were treated with solution of paraquat (1, 25, 50 and $100{\mu}M$ respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours. MIT and NR assay were performed to evaluate the cytotoxicity of cell organelles. $MTT_{50}\;and\;NR_{50}$ of paraquat were $1668.97{\mu}M\;and\;1030.85{\mu}M$, respectively. These $IC_{50}$ of Paraquat were decided as a low cytotoxicity by Borenfreund and Puemer (1984). In order to observe the toxicity and compensatory effects of paraquat on the rat lung, Spraque Dawley male rats were used as experimental animals and were divided into paraquat only treated group and simultaneous application group of paraquat and 3-MC, at 30 min and 1, 3, 6, 12, 24, 48 and 96 hrs interval after each treatment. The animals were sacrificed by decapitation and their or the lungs were immediately removed, immersed in fixatives, and were processed with routine method for light microscopic study. Paraffin sections were stained with H&E and iron hematoxylin of Verhoeff. Under the light microscopy, erythrocytes were full in alveolar capillaries at 3 hrs and congested at 24 hrs after paraquat administration. The great alveolar cells (Type II cell) were increased and mitosis of great alveolar were observed in interalveolar septa. Many lymphocytes, macrophages and polymorphonuclear (PMN) cells were observed in connective tissue surrounding lung tissue and germinal center in lymph follicles of terminal bronchiole. Alveolar macrophages were increased in interalveolar septa and alveoli at 48 hrs. And observed many alveolar macrophages at 96 hrs. In iron hematoxylin stain of Verhoeff, Collagen fiber were increased in respiratory bronchiole, interalveolar septa and alveoli and breath of alveoli, and alveolar pore were broaden. But, in paraquat plus 3-MC treated group, morphological changes were mild in lung tissue. These results indicate that 3-MC has a compensatory effects against toxicity of paraquat by conjugation with oxygen.