• Asian Pacific Journal of Cancer Prevention
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• 제16권2호
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• pp.741-746
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• 2015

Purpose: To investigate the influence of exogenous p53 upregulated modulator of apoptosis (PUMA) expression on cell proliferation and apoptosis in human non-small cell lung cancer A549 cells and transplanted tumor cell growth in nude mice. Materials and Methods: A549 cells were divided into the following groups: control, non-carrier (NC), PUMA (transfected with pCEP4-(HA) 2-PUMA plasmid), DDP ($10{\mu}g/mL$ cisplatin treatment) and PUMA+DDP (transfected with pCEP4-(HA)2-PUMA plasmid and $10{\mu}g/mL$ cisplatin treatment). The MTT method was used to detect the cell survival rate. Cell apoptosis rates were measured by flow cytometry, and PUMA, Bax and Bcl-2 protein expression levels were measured by Western blotting. Results: Compared to the control group, the PUMA, DDP and PUMA+DDP groups all had significantly decreased A549 cell proliferation (p<0.01), with the largest reduction in the PUMA+DDP group. Conversely, the apoptosis rates of the three groups were significantly increased (P<0.01), and the PUMA and DDP treatments were synergistic. Moreover, Bax protein levels significantly increased (p<0.01), while Bcl-2 protein levels significantly decreased (p<0.01). Finally, both the volume and the weights of transplanted tumors were significantly reduced (p<0.01), and the inhibition ratio of the PUMA+DDP group was significantly higher than in the single DDP or PUMA groups. Conclusions: Exogenous PUMA effectively inhibited lung cancer A549 cell proliferation and transplanted tumor growth by increasing Bax protein levels and reducing Bcl-2 protein levels.

• 한국균학회지
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• 제11권2호
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• pp.69-77
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• 1983

한국산(韓國産) 담자균류(擔子菌類)의 항암(抗癌) 성분(成分)을 탐색하기 위하여, 팽나무버섯의 자실체(字實體)를 온수(溫水)로 추출한 후, visking tube로 투석 및 냉동건조하여 단백성 다당류를 분리하였다. 이 성분을 ICR마우스에 이식한 sarcoma 180에 대한 종양억제작용을 실험하였던 바, 10mg/kg/day 용량(用量)을 10일간 투여한 군(群)에서 그 종양억제율이 대조군에 비하여 62.3%이었다. 처치군의 10마리중 3마리에서 그 종양이 완전히 퇴행하였다. 이 항암성분을 분석하였던 바, 42.4%의 다당류와 24.5%의 단백질을 함유하고 있었다. 그 다당류를 가수분해하여 분석하여 본 바, glucose, galactose, mannose, xylose 및 fucose의 5종의 단당류를 검출하였다. 그 단백질을 가수분해하여 16종의 아미노산을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제28권2호
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• pp.255-261
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• 2018

Aglycon protopanaxatriol (APPT) has valuable pharmacological effects such as memory enhancement and tumor inhibition. ${\beta}$-Glycosidase from the hyperthermophilic bacterium Dictyoglomus turgidum (DT-bgl) hydrolyzes the glucose residues linked to APPT, but not other glycoside residues. ${\beta}$-Glycosidase from the hyperthermophilic bacterium Pyrococcus furiosus (PF-bgl) hydrolyzes the outer sugar at C-6 but not the inner glucose at C-6 or the glucose at C-20. Thus, the combined use of DT-bgl and PF-bgl is expected to increase the biotransformation of PPT-type ginsenosides to APPT. We optimized the ratio of PF-bgl to DT-bgl, the concentrations of substrate and enzyme, and the reaction time to increase the biotransformation of ginsenoside Re and PPT-type ginsenosides in Panax ginseng leaf extract to APPT. DT-bgl combined with PF-bgl converted 1.0 mg/ml PPT-type ginsenosides in ginseng leaf extract to 0.58 mg/ml APPT without other ginsenosides, with a molar conversion of 100%. We achieved the complete biotransformation of ginsenoside Re and PPT-type ginsenosides in ginseng leaf extract to APPT by the combined use of two ${\beta}$-glycosidases, suggesting that discarded ginseng leaves can be used as a source of the valuable ginsenoside APPT. To the best of our knowledge, this is the first quantitative production of APPT using ginsenoside Re, and we report the highest concentration and productivity of APPT from ginseng extract to date.

• 한국균학회지
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• 제12권1호
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• pp.35-43
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• 1984

애기졸각버섯의 균사를 액내 진탕 배양법으로 배양하여 항종양성 성분을 생성시키고 이 성분의 특징을 밝히기위하여 위의 방법으로 배양된 균사체를 증류수로 가온 추출하였으며 그 추출물을 이온 교환수지 및 겔여과크로마토그라피법으로 정제하였다. 이 정제과정에서 분리된 5개의 분획에 대하여 각각 항종양 실험을 시행하였던 바, 그 정제의 정도에 비례하여 항암작용이 증가하였으며, 최종 분획 E를 1회 10mg/kg용량으로 백서에 투여하였을 때 75%의 종양 억제율을 나타내었으며, 다당류 81% 및 단백질 4%를 함유하고 있었다. 이 성분은 백서의 복강내 마크로훼이지의 수를 증가시켰으며 이 성분을 laccaran으로 명명한다.

• 한국식품영양학회지
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• 제32권6호
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• pp.651-661
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• 2019

Suaeda asparagoides (Miq.) is a salt marsh plant, long been prescribed in traditional medicine for the treatment of hypertension and liver toxification in Asian countries. The powder of S. asparagoides was added at the ratio of 0, 5, and 10%, respectively, of grain-type Meju to manufacture Doenjang in brine according to the salt concentration (8 and 12%). After 24 weeks of fermentation, the Doenjang samples were determined to have an anti-inflammatory effect on RAW 264.7 cells. Evaluation of the anti-inflammatory effect of Doenjang added S. asparagoides powder extracted using 80% EtOH, was performed to study the inhibition of pro-inflammatory factors such as NF-κB (nuclar factor κB), NO (nitric oxide), TNF-α (tumor necrosis factor alpha), IL-6 (interleukin-6), iNOS (inducible nitric oxide synthase), and COX-2 (cyclooxygenase-2) in lipopolysaccharide (LPS)-induced RAW 264.7 cells. The results showed that the Doenjang extracts reduced the production of NO, IL-6, COX-2, and iNOS increased in the LPS-stimulated RAW cell without cytotoxicity. In the case of the NF-κB and TNF-α there was no significant difference between the control and samples. In conclusion, these results suggest that Doenjang added with the S. asparagoides powder acts as functional fermented food with anti-inflammation effect.

• 한국균학회지
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• 제12권4호
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• pp.129-132
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• 1984

한국산 담자균류로부터 새로운 항종양 성분을 개발하기 위하여 표고의 일개 균주인 Lentinus edodes DMC7을 인공배지에서 진탕배양하여 그 균사 배양물을 얻었다. 그중 배양 여액으로부터 수용성 고분자 분획을 분리하여 LF-3라고 명명하였다. LF-3는 ICR 마우스에 이식된 Sarcoma 180세포에 대하여 항암작용을 나타내었으며 그 저지율은 53. 1%였다.

• 한국균학회지
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• 제20권4호
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• pp.324-336
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• 1992

영지버섯 Ganoderma lucidum과 잔나비걸상버섯 Ganoderna applanatum의 원형질체 융합균주 5개에 대하여 항암실험을 실시하여 그 중에서 다른 것에 비해 항암력이 높은 융합균주 F-2를 선발하였다. 그 항암성분을 조사하기 위하여 융합체의 균사를 액내 배양하고 열수추출물로부터 얻은 단백 다당체를 DEAE cellulose ion exchange column chromatography와 Sepharose CL-4B gel filtration을 이용하여 5가지 분획(Fr. I-V)으로 분리, 정제하여 모균주들과의 차이점을 비교, 분석하였다. 모균주와 분리한 각 성분들을 20mg/kg/day 용량으로 마우스의 복강에 투여하였을 때 sarcoma 180 고형암에 대하여 균주보다 융합체 성분(Fr. ll)의 종양억제율이 최고 1.5배로 증가되었다. 그리고 마우스의 면역에 영향을 미치는 항암성분을 연구한 결과, 대조군에 비해 활성화된 대식세포에서 분비되는 superoxide anion의 양을 1.2배, 비장세포중의 용혈반 형성 세포수를 4.3배로 증가시켰다. 화학분석에 의하면 이 성분은 glucose, galactose, mannose, fucose와 xylose로 구성된 다당류가 85.2%이며 15종의 아미노산으로 구성된 단백질이 0.39%이었고, hexosamine이 0.39%로 구성되었으며 분자량은 $5.6{\times}10^4$ dalton이었다.

• The Korean Journal of Physiology
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• 제1권1호
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• pp.33-41
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• 1967

The metabolic patterns of C-1 and C-6-carbon atoms of glucose were observed in the tissue homogenates of the Ehrlich ascites tumor tissue which was incubated for 3 hours in the Dubnuff metabolic shaking incubator. $C^{14}-1-and\;C^{14}-6-glucose$ were used as tracers. The glucose media in which tissue homogenate was incubated was kept at a concentration of 200mg% glucose of carrier and appropriate amount of $C^{14}-1-or\;C^{14}-6-tracer$. At the end of 3 hour incubation, respiratory $CO_2$ samples trapped by alkaline which is placed in the tenter well of incubation flask were analyzed for the total $CO_2$ production rates and their radioactivities. The tissue homogenate samples after incubation were analyzed for their concentrations of glucose, lactate, pyruvate and glycogen and calculations were made on the glucose consumption rate, pyruvate and lactate accumulation rates. The following results were obtained. Data obtained in each group are as follows: 1. In the tissue homogenate, which was incubated with $C^{14}-1-glucose as a substrate, total $CO_2$ production rate averaged $19.0{\pm}5.0{\mu}M/hr/gm$ and the mean specific activity of respiratory $CO_2$ was $840{\pm}296\;cpm/mgC.$ Relative specific activity (RSA) which means the fraction of $CO_2$ derived from medium $C^{14}-1-glucose$ to total $CO_2$ production rate was calculated by ratio of SA of respiratory $CO_2$ and medium $C^{14}-1-glucose.$ RSA was $14.3{\pm}5.0%,$ Accordingly actual $CO_2$ production rate from medium $C^{14}-1-glucose$ showed a mean value of $2.79{\pm}1.35\;{\mu}m$ of which amount was equivalent to the mean value of total glucose consumption rate $(RGDco_2)$, namely, $5.1{\pm}1.3%.$ Lactate and pyruvate appearance rates averaged $7.13{\pm}1.26\;and\;0.21{\pm}0.02{\mu}M/hr/gm,$ respectively. Assuming that these 3 carbon compounds appeared in the medium were derived from glucose, calculations were made that relative glucose disappearance rate into lactate $(RGD_L)$ was $38.0{\pm}5.4%\;and\;RGD_P$ was $1.23{\pm}0.03%.$ Therefore, about 43.3% of the total glucose consumed were accounted for by conversion into the respiratory $CO_2$, lactate and pyruvate. 2. In the second group, which was incubated with $C^{14}-1-glucose$ as a substrate, glucose consumption rate, lactate and pyruvate appearance rates showed almost the same order as the values of the $C^{14}-1-glucose$ substrate group. However, RSA was remarkably decreased showing a mean value of $1.02{\pm}0.13%.$ This fact means that the C-6 carbon of glucose take the minor part in the oxidative metabolism of glucose. The glycogen level in both substrate tissue homogenate showed less than 0.3% of tissue weight. These low value suggested that there was an inhibition of carbohydrate synthesis in the Ehrlich ascites tumor tissue. 3. The catabolic pathway of glucose in the tumor tissue were analyzed on the basis of Bloom's principle from the values of RSA. It was found that in the tumor tissue more than 90% of $CO_2$ derived from glucose were oxidized via the alternate pathway other than principal EMP-TCA cycle such as hexose monophosphate pathway (HMP). From the data described above, it was assumed that in the Ehrlich tumor tissue anaerobic glycolysis proceeds normally although, the oxidation of products of anaerobic glycolysis via the TCA cycle is inhibited resulting in the accumulation of lactate and almost all of oxidative energy from glucose is released by oxidative pathway such as HMP.

• 한국균학회지
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• 18권2호
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• pp.58-69
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• 1990

담자균류인 영지 Ganoderma lucidum의 자실체를 열수추출한 후 얻은 단백 다당체를 DEAE cellulose 이온교환수지와 Sepharose CL-4B 젤 여과법을 이용하여 분리 정제하였다. 정제한 분획들을 각각 CR, IN, IA, GL과 GH로 명명하였다. 각각의 성분을 20mg/kg/day 용량으로 투여하였을 때 sarcoma 180 고형암에 대하여 Fr. GL이 81%의 가장 높은 종양 억제을을 나타냈으며 그 분자량은 47,000 dalton이었다. 그 구성성분은 82% 다당류, 8% 단백질 및 0.9% hexosamine 이었다. 그 다당류를 구성하는 단당류를 분석한 결과 glucose, galactose, mannose, fucose순으로 함유되어 있었다. 항암작용의 기전을 밝히기 위하여 마우스의 면역에 미치는 항암 성분의 영향을 연구한 결과 활성화된 마크로파지에서 분비되는 superoxide anion 양을 1.6배로, 그리고 hemolytic plaque assay에서의 용혈반 형성 세포수를 6배로 각각 증가시켰다. 이 결과로 항암작용은 고형암에 직접적으로 세포독성을 나타내는 것이 아니라, 면역세포를 활성화 혹은 강화시키는 작용임을 알 수 있다.

• Journal of Periodontal and Implant Science
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• 제24권2호
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• pp.321-339
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• 1994

The cytokines released by osteoblasts induce bone resorption via the differentiation of osteoclast precursors. In this process, $interleukin-1{\beta}$($IL-1{\beta}$)-induced bone resorption is mediated by granulocyte macrophage-colony stimulation factor(GM-CSF), interleukin-6 (IL-6), and tumor necrosis factor ${\alpha}$($TNF-{\alpha}$) released from osteoblasts. Since these cytokines (GM-CSF, IL-6, $TNF-{\alpha}$) are produced by not only osteoblasts but also monocytes, and interleukin-10(I1-10) inhibits the secretion of these cytokines from monocytes, it may be speculated that IL 10 could modulate the production of GM-CSF, IL-6, and $TNF-{\alpha}$ by osteoblasts, then control $IL-1{\beta}-induced$ bone resorption. Therefore, the aims of the present study were to examine the effects of IL-10 on bone resorption. The sixten or seventeen-day pregnant ICR mice were injected with $^{45}Ca$ and sacrificed one day after injection. Then fetal mouse calvaria prelabeled with $^{45}Ca$ were dissected out. In order to confirm the degree of bone resorption, mouse calvaria were treated with Lipopolysaccharide(LPS), $TNF-{\alpha}$, $IL-1{\alpha}$, IL-8, $IL-1{\beta}$, and $IL-1{\alpha}$, Then, IL-10 and $interferon-{\gamma}$ ($IFN-{\gamma}$) were added to calvarial medium, in an attempt to evaluate the effect of $IL-1{\beta}-induced$ bone resorption. In addition, osteoclasts formation in bone marrow cell cultures, and the concentration of IL-6, $TNF-{\alpha}$, and GM-CSF produced from mouse calvarial cells were investigated in response to $IL-1{\beta}$ alone and simultaneously adding f $IL-1{\beta}$ and IL-10. The degree of bone resorption was expressed as the ratio of $^{45}Ca$ release(the treated/the control). The osteoclasts in bone marrow cultures were indentified by tartrate resistant acid phosphatase(TRAP) stain and the concentration of the cytokines was quantified using enzyme linked immunosorbent method. As results of these studies, bone resorption was induced by LPS(1 ng/ml ; the ratio of $^{45}Ca$ release, $1.14{\pm}0.07$). Also $IL-1{\beta}$(1 ng/ml), $IL-1{\alpha}$(1 ng/ml), and $TNF-{\alpha}$(1 ng/ml) resulted in bone resorption(the rations of $^{45}Ca$ release, $1.61{\pm}0.26$, $1.77{\pm}0.03$, $1.20{\pm}0.15$ respectively), but IL-8 did not(the ratio of $^{45}Ca$ release, $0.93{\pm}0.21$). The ratios of $^{45}Ca$ release in response to IL-10(400 ng/ml) and $IFN-{\gamma}$(100 ng/ml) were $1.24{\pm}0.12$ and $1.08{\pm}0.04$ respectively, hence these cytokines inhibited $IL-1{\beta}$(1 ng/ml)-induced bone resorption(the ratio of $^{45}Ca$ release $1.65{\pm}0.24$). While $IL-1{\beta}$(1 ng/ml) increased the number of TRAP positive multinulcleated cells in bone marrow cultures($20{\pm}11$), simultaneously adding $IL-1{\beta}$(1 ng/ml) and IL-10(400 ng/ml) decreased the number of these cells($2{\pm}2$). Nevertheless, IL-10(400 ng/ml) did not affect the IL-6, GM-CSF, and $TNF-{\alpha}$ secretion from $IL-1{\beta}$(1 ng/ml)-activated mouse calvarial cells. From the above results, it may be suggested that IL-10 inhibites $IL-1{\beta}-induced$ osteoclast differntiation and bone resorption. However, the inhibitory effect of IL-10 on the osteoclast formation seems to be mediated not by the reduction of IL-6, GM-CSF, and $TNF-{\alpha}$ production, but by other mechanisms.