• Maxillofacial Plastic and Reconstructive Surgery
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• 제27권2호
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• pp.103-109
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• 2005

In spite of the ongoing advances, standard therapies for oral cancer still has some limitations in efficacy and in ability to prolong survival rate of advanced disease and result in significant functional defect and severe cosmetic deformity. Currently gene therapy using tumor suppressor gene is considered as a potent candidate for new therapeutic approaches that can improve efficacy and reduce complications. The purpose of this research is to identify the role of adenoviral vector to transfer HCCS-1 tumor suppressor gene in oral cancer cells and to find out whether there is a possibility for it to serve in the field of gene therapy. The human SCC-25 cell line was used for transfection. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transduced with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the tranfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1). DNA extracted from Ad5CMV-HCCS-1 revealed HCCS-1 gene is incorporated. The transduction efficiencies were over than 50% of SCC-25 cells with a MOI of 2 and over 95% with a MOI of 50. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transduced SCC-25 cells. There was no or very low transcription HCCS-1 mRNA in wild and Ad5CMV-LacZ transduced SCC-25 cells. Cells transduced with Ad5CMV-HCCS-1 showed significant growth inhibition. By day 6, Ad5CMV-HCCS-1 treated cell count was decreased to 30% of mock-infected cells, while that of Ad5CMV-LacZ treated cells was 90% of mock-infected cells (p<0.05). Finally, these result suggest that the Ad5CMV-HCCS-1 has potential as a gene therapy tool for oral cancer.

• KSBB Journal
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• 제30권4호
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• pp.148-154
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• 2015

Onion (Allium cepa) is one of the flavonoids-rich materials in human diet and onion peel, which is the onion by-products, contains over 20 times more quercetin than the flesh. In this study, to examine the anti-inflammatory effects of onion peel hot water extract (OPHWE), the cell viability, nitric oxide (NO), pro-inflammatory cytokines, such as interluekin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and IL-$1{\beta}$, were measured using the murine macrophage cell line RAW 264.7 cells. The Balb/c mice were used for an in vivo acute toxicity test and ICR mice were used for measurement of inhibition effects of croton oil-induced mouse ear edema. As a result, NO levels decreased in a dose-dependent manner. The production of IL-6, TNF-${\alpha}$, and IL-$1{\beta}$ was suppressed by 38%, 41%, and 34% respectively, compared with that of the LPS only group, without any cytotoxicity. The edema formation in the ICR mouse ear was also reduced compared to that in control. Moreover, there were no mortalities occurred in mice administered 5,000 mg/kg body weight of OPHWE. These results suggest that OPHWE has considerable anti-inflammatory activities and can be regarded as a potent candidate material to treat inflammatory diseases.

• BMB Reports
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• 제37권2호
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• pp.185-191
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• 2004

Tissue transglutaminase (tTGase) regulates various biological processes, including extracellular matrix organization, cellular differentiation, and apoptosis. Here we report the protective role of tTGase in the cell death that is induced by the tumor necrosis factor $\alpha$ (TNF-$\alpha$) and ceramide, a product of the TNF-$\alpha$ signaling pathway, in human neuroblastoma SH-SY5Y cells. Treatment with retinoic acid (RA) induced the differentiation of the neuroblastoma cells with the formation of extended neurites. Immunostaining and Western blot analysis showed the tTGase expression by RA treatment. TNF-$\alpha$ or $C_2$ ceramide, a cell permeable ceramide analog, induced cell death in normal cells, but cell death was largely inhibited by the RA treatment. The inhibition of tTGase by the tTGase inhibitors, monodansylcadaverine and cystamine, eliminated the protective role of RA-treatment in the cell death that is caused by TNF-$\alpha$ or $C_2$-ceramide. In addition, the co-treatment of TNF-$\alpha$ and cycloheximide ecreased the protein level of tTGase and cell viability in the RA-treated cells, supporting the role of tTGase in the protection of cell death. DNA fragmentation was also induced by the co-treatment of TNF-$\alpha$ and cycloheximide. These results suggest that tTGase expressed by RA treatment plays an important role in the protection of cell death caused by TNF-$\alpha$ and ceramide.

• Journal of Ginseng Research
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• 제24권4호
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• pp.196-201
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• 2000

흰쥐 소뇌로부터 과립신경세포를 배양하여 NaCN으로 유도되는 신경세포손상에 대한 ginsenosides의 보호효과를 검토하였다. NaCN(I~10 M)을 배양된 세포에 1시간 동안 처리하면 농도 의존적으로 신경세포사를 일으켰다. Ginsenosides(Rb$_1$, Rc, Re, Ri, Rg$_1$)(0.5, 5 $\mu\textrm{g}$/ml를 세포에 전처치하면 10 mM NaCN으로 유도되는 세포사가 현저히 감소되었다. Rb$_1$과 Rc(5$\mu\textrm{g}$/ml)는 5 mM NaCN에 의하여 배양액 중으로 유리되는 glutamate의 증가를 현저히 억제하였으며, 1 mM N3CN에 의하여 유발되는 세포내 $Ca^{2+}$농도의 증가를 억제하였다. NaCN으로 유발되는 세포독성은 또한 MK-801, verapamil, NAME에 의하여도 억제되었다. 따라서, NaCN으로 유도되는 신경세포사는 glutamate release를 통한 NMDA수용체의 활성화와 그에 따른 $Ca^{2+}$의 세포내유입에 의한 것임을 알수 있고, ginsenosides, 특히 Rb$_1$과 Rc는 $Ca^{2+}$의 유입을 억제하므로서 NaCN에 의한 신경세포사를 억제하는 것으로 생각된다.

• Archives of Pharmacal Research
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• 제29권8호
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• pp.633-644
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• 2006

In last couple of decades the use of natural compounds like flavonoids as chemopreventive agents has gained much attention. Our current study focuses on identifying chemopreventive flavonoids and their mechanism of action on human prostate cancer cells. Human prostate cancer cells (PC3), stably transfected with activator protein 1 (AP-1) luciferase reporter gene were treated with four main classes of flavonoids namely flavonols, flavones, flavonones, and isoflavones. The maximum AP-1 luciferase induction of about 3 fold over control was observed with $20\;{\mu}M$ concentrations of quercetin, chrysin and genistein and $50\;{\mu}M$ concentration of kaempferol. At higher concentrations, most of the flavonoids demonstrated inhibition of AP-1 activity. The MTS assay for cell viability at 24 h showed that even at a very high concentration $(500\;{\mu}M)$, cell death was minimal for most of the flavonoids. To determine the role of MAPK pathway in the induction of AP-1 by flavonoids, Western blot of phospho MAPK proteins was performed. Four out of the eight flavonoids namely kaempferol, apigenin, genistein and naringenin were used for the Western Blot analysis. Induction of phospho-JNK and phospho-ERK activity was observed after two hour incubation of PC3-AP1 cells with flavonoids. However no induction of phospho-p38 activity was observed. Furthermore, pretreating the cells with specific inhibitors of JNK reduced the AP-1 luciferase activity that was induced by genistein while pretreatment with MEK inhibitor reduced the AP-1 luciferase activity induced by kaempferol. The pharmacological inhibitors did not affect the AP-1 luciferase activity induced by apigenin and naringenin. These results suggest the possible involvement of JNK pathway in genistein induced AP-1 activity while the ERK pathway seems to play an important role in kaempferol induced AP-1 activity.

• 한방안이비인후피부과학회지
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• 제25권3호
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• pp.1-12
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• 2012

Objectives : Fagopyrum esculentum(FE) has been used for removal of inflammation of internal organs and treatment of sore and ulcer by heat toxin in Korean herbal medicines. In this study, To investigated the protective effect of FE on allergic response, we determined whether FE inhibits allergic response. Methods : The effect of FE was analyzed by ELISA, RT-PCR and Western blot in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase, as a marker of degranulation, cytokne, and intracellular ROS and MAPK and NF-${\kappa}B$ signaling. Results : We found that FE suppressed ${\beta}$-hexosaminidase release, the production of IL-4 and TNF-${\alpha}$ and intracellular ROS level in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. FE also significantly inhibited cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, p38 and $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ signal transduction pathway. Conclusions : Our results indicate that FE protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and ROS via the suppression MAPK and NF-${\kappa}B$ of signal transduction. Abbrevations : FE, Fagopyrum esculentum; RBL-2H3, rat basophilic leukemia cell line; ROS, reactive oxygen species; MAPK, Mitogen-activated protein kinase; $NF{\kappa}B$, nuclear factor ${\kappa}B$; $TNF{\alpha}$, Tumor necrosis factor alpha; GM-CSF, Granulocyte macrophage colony-stimulating factor; ERK, extracellular-signal-regulated kinase; JNK, c-Jun NH2-terminal kinase; p38, p38 MAP kinase; $I{\kappa}B{\alpha}$, inhibitory-kappa B alpha.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1988년도 학술대회지
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• pp.115-121
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• 1988

인삼에서 분리한 항염성분이 일찍이 파낙스 사포닌(${\beta}-{\beta}'$(Rc는 아님)는 chemotaxis에 억제효과($27.6-42.1\%$)를 나타내었는데 이는 스테로이드계 항염증제인 DXM으로 관찰한 것과 같거나 또는 더 강력하였다.($29.6\%$). DXM이 PMNL 화학발광을 억제하는 반면에 중성 사포닌에서는 효과가 관찰되지않았다. 인삼뿌리에서 추출한 몇 종류의 중성 dammarane계 사포닌이 PMNL 기능을 조절하는 효과가 있었으며 그 효과는 항염증제인 dexamethasone 과는 작용면에서는 다른 것으로 사료된다.

• 생명과학회지
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• 제17권11호
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• pp.1596-1600
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• 2007

본 연구에서 봉독의 apoptosis 유도에 의한 항암기전 효능을 A549 인체폐암세포를 대상으로 조사하였으며, apoptosis 유발에 관여할 것으로 예상되는 중요한 유전자들의 발현 및 활성에 미치는 봉독의 영향을 조사하였다. A549 세포의 생존율은 봉독의 처리 농도 증가에 따라 강력하게 억제되었으며, 이는 염색질 응축 현상을 동반한 apoptosis 유발에 의한 것임을 알 수 있었다. 봉독 처리에 의한 apoptosis 유발은 Bax 및 Bcl-xL의 발현 변화 없이 Bcl-2의 발현 감소에 따른 상대적인 Bax의 발현 증가와 IAP family에 속하는 인자들의 발현 감소 및 caspase의 활성화와 연관성이 있었다.

• 미생물학회지
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• 제42권3호
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• pp.165-171
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• 2006

ERM protein 은 23S rRNA의 $A_{2058}$에 dimethylation시킴으로써 $MLS_B$계 항생제의 부착을 저해하여 항생제의 활성을 억제하는 내생인자 단백질이다. ERM 단백질의 하나인 ErmSF의 N-말단부위(N-termimal end region, NTER)에 존재하는 1-25번째 아미노산을 함유하는 펩타이드의 활성에서의 역할을 알아보기 위해 이률 제거한 변이 단백질을 표현하는 유전자를 클로닝하고 대장균에서 수용성 단백질로 12.65 mg/L culture의 수율로 대량생산하였다. 이렇게 대량생산된 단백질의 활성을 in vivo와 in vitro에서 확인하였다. 그 결과 in vitro에서 야생형(wild type)의 단백질에 비해 15%의 활성이 감소한 것을 확인하였고 이는 제거된 펩타이드가 기질과 상호작용하여 효소의 활성에 영향을 미친다는 것을 시사하고 있다. 이렇게 감소된 효소의 활성은 생체 내(in vivo) 활성에도 적용되어 처음에는 변이 단백질을 함유하는 세포가 항생제의 작용에 의하여 성장억제를 받지만 시간의 경과와 함께 내성을 회복하여 밤샘 배양하였을 경우는 야생형 단백질을 함유한 세포와 동일한 내성 즉 항생제에 의한 성장억제지역(inhibition zone)을 전혀 나타내지 않는 것으로 밝혀졌다.

• 생명과학회지
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• 제15권4호
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• pp.626-632
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• 2005

폴리아민은 모든 진핵세포에서 발견되는 다가 양이온성의 저분자 물질이며 세포성장에 필수적인 것으로 알려져 있다. 본 논문에서는 폴리아민의 역할 중에서 산화적인 스트레스에 대한 세포보호 효과를 연구하였다. 쥐의 간세포주인 $Ac_2F$에 산화 스트레스를 유발하기 위하여 2,2'-azobis(2-amidinopropane)dehydrochloride (AAPH)를 처리하였을 때, 세포증식은 농도 의존적으로 감소하였다. 배지에 폴리아민을 첨가하였을 때 세포성장은 농도 의존적으로 증가하였으며 ROS 발생은 현저히 감소하였다. 폴리아민 가운데 특히 spermidine과 spermine이 뚜렷한 세포증식효과를 보였다. Spermine의 경우, $20{\mu}M$농도에서 AAPH에 의해 유도된 ROS발생을 $45\%$나 감소시켰다. 산화 스트레스에 관여하는 효소들 가운데 주된 효소인 superoxide dismutate (SOD)와 catalase (CAT)의 세포 내 단백질을 Western blotting으로 조사한 결과, AAPH는 이 두 가지 단백질의 생성을 억제한 것으로 나타났다. 그러나 spermine을 처리하였을 때 두 단백질의 생산은 모두 정상적으로 회복이 되었다. 또한 세포주기의 중요한 조절 단백질인 cyclin E 역시 AAPH에 의하여 생성이 억제되었다. 이는 AAPH에 의하여 생성된 ROS가 세포주기의 S phase의 진행을 억제한 것으로 생각된다. AAPH에 의한 cyclin E의 억제는 spermine에 의하여 정상적으로 회복되었다. 위와 같은 Spermine의 항산화 효과는 ethidium bromide와 acridine orange를 이용하여 형태학적으로도 증명되었다.