• Nutrition Research and Practice
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• 제9권2호
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• pp.111-116
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• 2015

BACKGROUND/OBJECTIVES: Inonotus obliquus (I. obliquus, Chaga mushroom) has long been used as a folk medicine to treat cancer. In the present study, we examined whether or not ethanol extract of I. obliquus (EEIO) inhibits cell cycle progression in HT-29 human colon cancer cells, in addition to its mechanism of action. MATERIALS/METHODS: To examine the effects of Inonotus obliquus on the cell cycle progression and the molecular mechanism in colon cancer cells, HT-29 human colon cancer cells were cultured in the presence of $2.5-10{\mu}g/mL$ of EEIO, and analyzed the cell cycle arrest by flow cytometry and the cell cycle controlling protein expression by Western blotting. RESULTS: Treatment cells with $2.5-10{\mu}g/mL$ of EEIO reduced viable HT-29 cell numbers and DNA synthesis, increased the percentage of cells in $G_1$ phase, decreased protein expression of CDK2, CDK4, and cyclin D1, increased expression of p21, p27, and p53, and inhibited phosphorylation of Rb and E2F1 expression. Among I. obliquus fractions, fraction 2 (fractionated by dichloromethane from EEIO) showed the same effect as EEIO treatment on cell proliferation and cell cycle-related protein levels. CONCLUSIONS: These results demonstrate that fraction 2 is the major fraction that induces $G_1$ arrest and inhibits cell proliferation, suggesting I. obliquus could be used as a natural anti-cancer ingredient in the food and/or pharmaceutical industry.

• Asian Pacific Journal of Cancer Prevention
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• 제13권4호
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• pp.1333-1340
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• 2012

Epithelial ovarian cancer represents the most lethal gynecological cancer, and the high mortality rate makes this malignancy a major health concern. Poor prognosis results from an inability to detect ovarian cancers at an early, curable stage, as well as from the lack of an effective therapy. Thus, effective and novel strategies for prevention and treatment with non-toxic agents merit serious consideration. Resveratrol, obtained from grapes, berries, peanuts and red wine, has been shown to have a potent growth-inhibitory effect against various human cancer cells as well as in in vivo preclinical cancer models. The objective here was to evaluate potential antitumor effects of resveratrol in both in vitro and in vivo NuTu-19 ovarian cancer models. In vitro an invasion assay was performed. After 48 h, the numbers of viable cells that invaded the extracellular matrix layer were reduced by 94% with resveratrol in comparison to control. For the in vivo anti-tumor assessment, 10 rats were injected with NuTu-19 cells into the ovarian bursa. Thereafter, half were provided with a diet mixed with a dose of 100 mg resveratrol/kg body weight/day for 28 days. Following sacrifice, anticancer effects were assessed by histological evaluation of ovarian as well as surrounding tissues, and immunohistochemical detection of cell proliferation and apoptosis, but there were no observable differences between the control and resveratrol-treated groups for any of the biological endpoints. While resveratrol is effective in suppressing the in vitro cellular invasion of NuTu-19 ovarian cancer cells, these effects do not appear to impact on in vivo NuTu-19 ovarian cancers in rats.

• 동의생리병리학회지
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• 제22권2호
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• pp.282-289
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• 2008

We evaluated the effect of SHBCS on adhesion and invasion of colon L5-26 adenocarcinoma cell line in vitro in vitro and experimental liver metastasis in vivo. SHBCS showed little inhibitory effect on colon 26-L5 cell proliferation. At the concentration of up to 500 mg/ml of SHBCS 80% of cells were viable. SHBCS showed no inhibitory effect on adhesion and invasion of colon 26-L5 cells, which were placed on matrigel. In a dose dependent manner, oral administration of SHBCS showed a significantly inhibitory effect on liver metastasis from colon 26-L5 injected mice. When mice were depleted of NK cells or macrophages before tumor inoculation, SHBCS significantly decreased liver metastasis fromf the tumor injected mice. Compared with the control mice, SHBCS increased the populations of macrophages and NK cells by 30%, 18%(10 mg/mouse, 50 mg/mouse) and 5%, 1% (10 mg/mouse, 50 mg/mouse) respectively. Compared with the control mice, SHBCS increased the populations of CD4 cells by 5%, 18% (10 mg/mouse, 50 mg/mouse) respectively. Spelenocytes from mice administerd with SHBCS were stimulated with LPS plus ConA, proliferation of splenocytes from mice administerd with SHBCS was 140%, 146%(10 mg/mouse, 50 mg/mouse) compared with th control mice. In conclusion, the present study suggests that SHBCS may have an inhibitory effect on liver metastasis through immunopotentiating activity which is associated with macrophages and NK cells.

• 한국수의병리학회지
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• 제2권2호
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• pp.127-138
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• 1998

Transmissible venereal tumor(TVT) is a naturally occurring contagious neoplasm which can be transmitted by mechanical contact during mating in dogs and transplanted as intact viable cells to dogs and other members of canine family such as coyotes, jackals, wolves, and foxes. The incidence of this tumors tends to increase in Korean native Jindo dogs. This is probably due to the high density and unrestrained management system. With time, TVT reaches the maximum size and then tends to regress spontaneously unless individuals are immunologically compromised. It consists of different types of cells depending on the stage. In this study, 10 tumors were selected from Jindo dogs. These were histologically calssified into three stages; progressive, steady-state, and regressive. Mitotic figures were counted, and their histological appearance at each stage is compared with their DNA ploidy. Histologically, 5 tumor cases were calssed as the progressors, 3 cases as the steady-state tumors, and 2 cases as regressors. Progressors were composed of round cells with large nuclei containing conspicuous nucleoli and frequent mitotic figures. A few spindle-shaped cells and inflammatory cells including mainly lymphocytes, a few neutrophils and macrophages were also seen. In the steady-state tumors, there was an increased number of spindle shaped cells and mitotic figures were rare. Six tumors were diploid and four were aneuploid with the variation coefficient of 7.02. Two of five progressive tumors were aneuploid. Two of three steady-state tumors were aneuploid while both tumors at the regressive stage were diploid. Progressive and steady-state tumors had a much larger S/G2M fraction and a higher mitotic index than regressive tumors. Two tumors which persisted for more than one year were aneuploid. These results suggest that the progressive and steady-state tumors had more active cell division than the regressive neoplasms.

• 대한미생물학회지
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• 제22권4호
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• pp.435-443
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• 1987

A total of 30 strains of Candida albicans were examined for susceptibility to amphotericin B and ketoconazole using Sabouraud's dextrose broth, Kimmig broth and Supplemented yeast nitrogen base broth media. Furthermore, the growth curve and colony forming units were checked for use of stationary-phase cells and 2-hour incubation cells in the absence of atifungal agents. The viable counts were determined periodically during incubation by standard plate count techniques. The minimum inhibitory concentrations of amphotericin B for use of stationary phase cells were as follows: SDB, $0.09{\sim}0.97mcg/ml$(0.39mcg/ml); Kimmig broth, $0.19{\sim}0.39mcg/ml$(0.42 mcg/ml) and SYNB, $0.19{\sim}0.39mcg/ml$mcg/ml(0.23mcg/ml). In ketoconazole, MICs were value SDB, $3.12{\sim}25.0mcg/ml$(12.5mcg/ml); Kimmig broth, $12.5{\sim}25.0mcg/ml$ (22.5mcg/ml) and SYNB, $3.12{\sim}12.5mcg/ml$(6.71mcg/ml). The MICs of amphotericin B(0.2mcg/ml cone.) for use of 2-hour incubation cells in absence of AMB were, SDB, $0.04{\sim}0.39mcg/ml$(0.11mcg/ml); Kimmig broth, $0.09{\sim}0.39mcg/ml$(0.18mcg/ml) and SYNB, $0.09{\sim}0.19mcg/ml$(0.14mcg/ml) and in KTZ, the value of MICs were SDB, $3.12{\sim}25.0mcg/ml$(12.22mcg/ml); Kimmig broth, $0.78{\sim}25.0mcg/ml$(11.01mcg/ml) and SYNB, $1.56{\sim}12.5mcg/ml$(3.90mcg/ml). The two-log reductions in CFU per milliliter observed when 2 hour preincubation cells were treated with 0.2mcg/concentrations of AMB and 25.0mcg/ml of KTZ. However, AMB treated cells were restored to growth activity, it suggested that the AMB has no active antifungal activity.

• Journal of Acupuncture Research
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• 제36권3호
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• pp.140-146
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• 2019

Background: This study investigated the effects of Vipera lebetina turanica snake venom (SV) on cerebral infarction induced by middle cerebral artery occlusion in mice. Methods: Following cerebral infarction, SV was injected intravenously or added to BV2 cell culture. Tissue injury was detected using triphenyltetrazolium chloride (TTC) staining, neurological deficit score, NO, ROS, and GSH/GSSG assays, qPCR, Western blot, and cell viability. Results: Cerebral infarction caused by middle cerebral artery occlusion as observed by TTC staining, showed SV inhibited cell death, reducing the number of brain cells injured due to infarction. SV treatment for cerebral infarction showed a significant decrease in abnormal behavior, as determined by the neurological deficit score. The oxidation and inflammation of the cells that had cerebral infarction caused by middle cerebral artery occlusion (NO assay, ROS, GSH/GSSG assay, and qPCR), showed significant protection by SV. Western blot of brain infarction cells showed the expression of iNOS, COX-2, p-IkB-${\alpha}$, P38, p-JNK, p-ERK to be lower in the SV group. In addition, the expression of IkB increased. BV2 cells were viable when treated with SV at $20{\mu}g/mL$ or less. Western blot of BV2 cells, treated with 0.625, 1.5, $2.5{\mu}g/mL$ of SV, showed a significant decrease in the expression of p-IkB-${\alpha}$, p-JNK, iNOS, and COX-2 on BV2 cells induced by LPS. Conclusion: SV showed anti-inflammatory and anti-oxidant effects against cerebral infarction and inflammation.

• Journal of Veterinary Science
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• 제23권4호
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• pp.47.1-47.11
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• 2022

Background: In lipopolysaccharide-induced RAW264.7 cells, Aster tataricus (AT) inhibits the nuclear factor kappa-light-chain-enhancer of activated B cells and MAPKs pathways and critical pathways of osteoclast development and bone resorption. Objectives: This study examined how aster saponin A2 (AS-A2) isolated from AT affects the processes and function of osteoclastogenesis induced by receptor activator of nuclear factor kappa-B ligand (RANKL) in RAW264.7 cells and bone marrow macrophages (BMMs). Methods: The cell viability, tartrate-resistant acid phosphatase staining, pit formation assay, polymerase chain reaction, and western blot were carried out to determine the effects of AS-A2 on osteoclastogenesis. Results: In RAW264.7 and BMMs, AS-A2 decreased RANKL-initiated osteoclast differentiation in a concentration-dependent manner. In AS-A2-treated cells, the phosphorylation of ERK1/2, JNK, and p38 protein expression were reduced considerably compared to the control cells. In RAW264.7 cells, AS-A2 suppressed the RANKL-induced activation of osteoclast-related genes. During osteoclast differentiation, AS-A2 suppressed the transcriptional and translational expression of NFATc1 and c-Fos. AS-A2 inhibited osteoclast development, reducing the size of the bone resorption pit area. Conclusion: AS-A2 isolated from AT appears to be a viable therapeutic therapy for osteolytic illnesses, such as osteoporosis, Paget's disease, and osteogenesis imperfecta.

• Journal of Photoscience
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• 제9권2호
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• pp.225-228
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• 2002

Bcl-2 is a member of large bcl-2 family and protects cells from apoptosis. Using bcl-2-expressing adenovirus vector (Ad-bc1-2), we investigated the effect of bc1-2 on UVB-induced apoptosis. Adenovirus vector efficiently introduced bc1-2 gene in cultured normal mouse keratinocytes (NMK cells); almost all NMK cells (lx10$^{6}$ ) were transfected at Ixl0$^{8}$ PFU/ml. Bcl-2-transfected NMK cells were significantly resistant to UVB-induced apoptosis with the suppressive effect dependent on bcl-2-expression level. Following UVB irradiation caspase 8, 3, 9 activities were stimulated in NMK cells, while in bc1-2-transfected cells, only caspase 8, but not caspase 3 or 9 activities were stimulated. In order to investigate the effect of bc1-2 in vivo, topical application of Ad-bc1-2 on tape-stripped mouse skin was performed. Following the application, Bc1-2 was efficiently overexpressed in almost all viable keratinocytes. The expression was transient with the maximal expression of Bc1-2 at 1st day following the application of lxl0$^{9}$ PFU in 200ml. The introduced Bc1-2 remained at least for 6 days. UVB irradiation (1250 J/m$^2$) induced apoptosis within 12 h and the maximal effect was observed at 24 h in control mouse skin. Bc1-2-transfected mice skin were resistant to UVB-induced apoptosis. Topical application of empty adenovirus vector alone had no effect on Bc1-2 expression or UVB-induced apoptosis. These results indicate that adenovirus vector is an efficient gene delivery system into keratinocytes and that Bcl-2 is a potent inhibitor of UVB-induced apoptosis both in vitro and in vivo.

• Toxicological Research
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• 제11권1호
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• pp.1-8
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• 1995

Fumonisins are a family of mycotoxins produced by the fungus Fusarium moniliforme which is a common contaminant in corn. Fumonisins are potent inhibitors of sphingosine and sphinganine N-acyltransferase (ceramide synthase), key enzymes in sphingolipid metabolism. The purpose of this study was to provide the evidence that the elevated levels of free sphingoid bases (primarily sphinganine) and depletion of complex sphingolipids were closely related to the inhibition of cell growth in LLC-$PK_1$ cells exposed to fumonisin $B_1$$(\leq 35 {\mu}M)$. Concentrations of fumonisin $B_1$ between 10 and $35 {\mu}M$ were known to inhibit cell growth without cytotoxicity in $LLC-PK_1$ cells (Yoo et al. Toxicol. Appl. Pharmacol. 114, 9-15, 1992). Cells exposed to 35$\mu M$ fumonisin B$_1$ for 48 and 72 hr developed a fibroblast-like (elongated and spindle-shaped) appearance and were less confluent than normal cells. At between 24 and 48 hr after exposure to fumonisin $B_1$ cells were beginning to show the inhibition of cell growth and at 72 hr the number of viable cells in fumonisin-treated cultures was about 50% of concurrent control cultures. During the 24 hr lag period preceding inhibition of cell growth, the free sphinganine levels in cells exposed to $35 {\mu}M$ fumonisin $B_1$ were highly elevated (approximately 230 fold higher than normal cells). The elevated levels of free sphinganine were $435\pm14$$pmoles/{10^6}$ cells at 48 hr and approximately TEX>$333\pm11$$pmoles/{10^6}$ cells in cells exposed to $35{\mu}M$ fumonisin$B_1$ at 72 hr, while the levels of free sphinganine in normal cells were less than 2$pmoles/{10^6}$ cells. Under the same condition, depletion of intracellular complex sphingolipids as a consequence of fumonisin inhibition of de novo sphingolipid biosynthesis and turnover pathway was appeared. Content of free sphingold bases in dividing cells was more elevated than in confluent cells at 24-48 hr after cells were exposed to $20{\mu}M$ fumonisin $B_1$. The dividing cells were showing the inhibition of cell growth at 48-72 hr and $20{\mu}M$ fumonisin $B_1$. The results of this study support the hypothesis that the inhibition of cell growth is very well related to the disruption of sphingolipid metabolism in $LLC-PK_1$ cells.

• 한국식품영양학회지
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• 제15권1호
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• pp.42-50
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• 2002

Skim milk에 매실 착즙액을 수준별로 첨가하고 4종(Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei)의 젖산균을 단독균주 또는 혼합균주로 접종하여 매실 착즙액의 첨가가 젖산균의 산생성 및 생육에 미치는 영향과 저장성을 조사하였고, 매실 착즙액으로 사용한 매실의 일반 성분을 분석하였다. 매실의 일반 성분은 조회분 0.4%, 식이섬유 4.1%, 구연산 4.66%, 총당 0.264%, vitamin C 405.34mg%이었고, 매실 착즙액의 첨가는 젖산균의 증식을 촉진시켰으며 산생성도 증가하였다. 실험구 중 Str. thermophilus, Lac. acidophilus와 Lac. casei의 혼합균주에 3% 매실 착즙액을 첨가한 실험구가 가장 많은 양의 젖산(1023%)을 생성하였고, 가장 높은 생성균수(3.6$\times$$10^{11}$ cfu/mL)를 나타내었다. 매실 착즙액 3% 첨가 호상 요구르트를 대조구와 함께 4$^{\circ}C$와 2$0^{\circ}C$에서 30일 동안 저장한 결과, 세 개의 혼합균주(Str. thermophilus, Lac. acidophilus)를 사용한 호상 요구르트가 4$^{\circ}C$에서 pH는 4.25, 생균수는 4.1$\times$$10^{9}$ cfu/mL 이었고, 2$0^{\circ}C$에서 pH는 3.76, 생균수는 8.7$\times$$10^{8}$ cfu/mL으로 대조구인 매실 무첨가 호상 요구르트에 비하여 높은 저장성을 나타내었다. 이상의 결과로 보아 매실 착즙액을 첨가한 호상 요구르트의 제조가 가능할 것으로 사료된다.