• 생명과학회지
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• 제24권1호
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• pp.1-7
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• 2014

진핵세포의 유전자 발현은 genomic DNA 부위의 프로모터라고 불리우는 지역에 전사인자와 RNA 중합효소가 자리하면서 시작되는 기작이다. 유전자 내의 프로모터를 동정하는 여러종류의 실험 방법들이 있지만, 많은 시간과 노동력이 요구되어진다. 본 연구에서는 Ensembl, NCBI, CpG plot 등과 같은 생물정보학 관련 프로그램들을 활용하여 SETDB1 유전자의 프로모터를 동정하여 클로닝하고자 하였다. PCR 증폭을 수행한 후 얻은 약 2 kb DNA 조각을 SETDB1-P1이라 명명하였으며, PCR 산물은 TA 벡터로 클로닝 후 확인하였으며, 이를 다시 제한 효소 절단을 통하여 pGL3-luc 벡터로 클로닝하였다. 클로닝된 pGL3-SETDB1-P1-luc 플라스미드를 H1299 폐암세포주에 transfection 시킨 후 여러 가지 항암제를 처리하였을 때, taxol, 5-FU, doxorubicin 처리군에서 SETDB1 프로모터 활성이 감소하는 것을 확인하였다. 이러한 결과는 웨스턴 블롯 및 RT-PCR 실험을 통해 항암제 처리 후 SETDB1 유전자 발현이 조절됨을 확인하였다. 그러므로 bioinformatics 프로그램을 통한 프로모터 동정 및 클로닝 방법을 다른 유전자들에도 적용시킨다면, 유전자 발현 연구에 매우 유용할 것으로 사료된다.

• KSBB Journal
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• 제13권2호
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• pp.174-180
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• 1998

유럽주목(Taxus baccata Pendula)현탁 세포 배양에서(10-deacetyl) baccatin III 생산을 증진시키는 연구를 하였다. 높은 초기 당 농도가(10-deacetyl) baccain III 생산을 증진시키는 연구를 하였다. 6% 의 초기 포도당과 자당의 경우 1O-deacetyl baccatin III 생산을 각각 3.배와 2.5배씩 증가시켰다. Baccain III도 초기 포도당 8%에서,초기 자당 6%에서 최대값을 보였다. 일리시터로서 methyl jasmonate는(10-deacetyl) baccatin III 의 생성은 $50{\mu}$M에서 최대치를 보였는데 이는 대조구에 비하여 4.5배 증가한 값이었다.Baccatin III 의 경우 methyl jasmonate $100{\mu}$M에서 생성된 Baccatin III 최대치는 대조구에 비하여 7.배 증가한 값이었다. Methyl jasmonate elicitation 에 의한(1O-deacetyl) baccatin III.과 각 taxane의 시간별 생성 경향을 보면 baccatin III와 10-deacetyl baccatin III는 투여 후 1일 까지 생성이 증가하다 그 후부터 감소하였고, paclitaxel, 10-deacetyl taxol 및 cephalomanine 의 생성이 이어졌다. (10-deacetyI) baccatin III의 생합성을 증가시키기 위하여 전구 물질을 투여하였다. Benzoic acid를 $500{\mu}M$로 투여 하였을 때 10-deacetyl baccatin III 및 baccatin III의 생산은 대조구의 비하여 각각 10배와 13배 증가 하였다. Lysine도 $500{\mu}M$로 투여하였을 때 baccatin III 의 생성량을 대조구 보다 8배 증가시켰다. 증가시켰다.

• 대한한의학회지
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• 제34권3호
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• pp.126-142
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• 2013

Objectives: Previous reports have shown that Bogijetong-Tang (BJT) is effective in peripheral neuropathy induced by taxol and crush injury. In this study, we researched the effects of BJT on diabetic neuropathy induced by STZ in the mouse. Methods: We performed both in vitro and in vivo experiments to verify the effects of BJT on diabetic neuropathy induced by STZ in mice. Changes in axonal recovery were observed with immunofluorescence staining using NF-200, Hoechst33258, $S100{\beta}$, caspase 3 and anti-cdc2. Proliferation and degeneration of Schwann cells were investigated by immunofluorescence staining and western blot analyses. Results: BJT showed considerable effects on neurite outgrowth and axonal regeneration in diabetic neuropathy. BJT contributed to the creation of NF-200, GAP-43, Cdc2, phospho-vimentin, ${\beta}1$, active ${\beta}1$, ${\beta}3$ integrin, phospho-Erk1/2 protein. Conclusions: Through this study, we found that BJT is effective for enhanced axonal regeneration via dynamic regulation of regeneration-associated proteins. Therefore, BJT had a pharmaceutical property enhancing recovery of peripheral nerves induced by diabetic neuropathy and could be a candidate for drug development after more research.

• Asian Pacific Journal of Cancer Prevention
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• 제13권2호
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• pp.563-567
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• 2012

In our era there has been several anti-cancer drugs which have undergone both experimental and clinical trials; however, due to their poor solubilities, numerous side effects, insufficient bioavailability and poor compliance, many have resulted into poor outcomes. Therefore, our aim was to investigate the effects of novel hydrophilic taxanes analogues CQMU-0517 and CQMU-0519 on growth of A549 lung, SKVO3 ovary and MCF7 breast carcinoma cell lines. Different concentrations of original paclitaxel, CQMU-0517, original docetaxel and CQMU-0519 were utilized on three cell lines, where cell growth was assessed using cell culture kit-8 and flow cytometry analysis. The results unveiled that CQMU-0517 and CQMU-0519 suppressed cell growth in the three particular cell lines, cell cycle arrest being evident in the G2/M phase. Hence, the results showed that these new taxane analogues have potential and warrant future clinical trials.

• BMB Reports
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• 제49권4호
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• pp.238-243
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• 2016

The efficacy of anticancer drugs depends on a variety of signaling pathways, which can be positively or negatively regulated. In this study, we show that SETDB1 HMTase is down-regulated at the transcriptional level by several anticancer drugs, due to its inherent instability. Using RNA sequence analysis, we identified FosB as being regulated by SETDB1 during anticancer drug therapy. FosB expression was increased by treatment with doxorubicin, taxol and siSETDB1. Moreover, FosB was associated with an increased rate of proliferation. Combinatory transfection of siFosB and siSETDB1 was slightly increased compared to transfection of siFosB. Furthermore, FosB was regulated by multiple kinase pathways. ChIP analysis showed that SETDB1 and H3K9me3 interact with a specific region of the FosB promoter. These results suggest that SETDB1-mediated FosB expression is a common molecular phenomenon, and might be a novel pathway responsible for the increase in cell proliferation that frequently occurs during anticancer drug therapy.

• 생약학회지
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• 제26권4호
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• pp.344-348
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• 1995

Preliminary screening test of modulators for multidrug resistance with 400 medicinal plants was carried out by using human multidrug resistance cell line, KB-V1. Among active medicinal plants, the unripe fruits of Evodia officinalis showed a potent modulating activity of MDR. From MeOH extract of this plant, we isolated two indole alkaloids, rutaecarpine (1) and evodiamine (2), by repeated silicagel column chromatography. Rutaecarpine increased the cytotoxicities of vinblastine and taxol against multidrug resistance cells, but evodiamine showed no modulating activity in spite of its potent cytotoxic activities.

• Asian Pacific Journal of Cancer Prevention
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• 제16권13호
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• pp.5169-5173
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• 2015

To determine the efficacy of postoperative adjuvant chemotherapy with paclitaxel plus cisplatin (Taxol + DDP, TP therapy) for stage IIA esophageal squamous cell carcinoma (ESCC) and to investigate the expression of RUNX3 in lymph node metastasis-negative esophageal cancer and its relationship with medical prognosis, a retrospective summary of clinical treatment of 143 cases of stage IIA esophageal squamous cell carcinoma patients was made. The patients were divided into two groups, a surgery alone control group (52 patients) and a chemotherapy group that received postoperative TP therapy (91 patients). The disease-free and 5 year survival rates were compared between the groups and a multivariate analysis of prognostic factors was performed. The same analysis was performed for cases classified as RUNX3 positive and negative, with post-operative specimens assessed by immunohistochemistry. Although the disease-free and 5 year survival rates in control and chemotherapy groups did not significantly differ and there was no significance in RUNX3 negative cases, postoperative adjuvant chemotherapy in the chemotherapy group was shown to improve disease-free and 5 year survival rate compared to the control group in RUNX3 positive cases. On Cox regression multivariate analysis, postoperative adjuvant chemotherapy (P<0.01) was an independent prognostic factor for RUNX3 positive cases, suggesting that postoperative TP may be effective as adjuvant chemotherapy for stage IIA esophageal cancer patients with RUNX3 positive lesions.

• KSBB Journal
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• 제14권4호
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• pp.408-413
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• 1999

빠른 분석시간과 우수한 분리도를 가진 분석기기로서 크로마토그래피가 분취용으로 이용되기 위해서는 많은 시료양을 취급할 수 있어야 한다. 이 경우 충전물과 관의 크기가 커지게 되면 관의 효율은 분석용에 비해서 떨어지게 된다. 순도와 수율에 미치는 변수가 서로 복잡하게 연관되어 있기때문에 특히 분취용 조업조건의 최적화는 많은 경험과 실험이 요구된다. 또한 많은 상관된 실험변수들이 있고 분리하고자 하는 물질의 종류가 다양하기 때문에 수학적 모델을 적용하여 시료가 관내에서 거동 (behavior)을 예측하는 것이 쉽지 않다. 이러한 문제점을 해결하기 위해서 외국에서는 최소한의 실험을 통하여 고분가 가치 물질을 효율적이고 경제적으로 얻기 위한 크로마토그래피용 프로그램을 활발하게 이용하기 시작하였다. 인하대학교 고순도분리연구실에서 개발된 HCI 프로그램은 단지 LC에 국한된 것이 아니라 GC,SFC (supercritical fluid chromatography) 뿐만아니라 연속식 시스템인 SMB (simulated moving bed)까지 확장될 예정이다.

• Animal cells and systems
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• 제1권1호
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• pp.63-69
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• 1997

We have previously reported that NF-kB is involved in the regulation of nitric oxide synthase gene expression during differentiation of chick embryonic myoblasts. However, how NF-kB is timely activated during myogenesis remains elusive. One of the most prominent events in myogenesis is myoblast membrane fusion, which is accompanied with massive cytoskeletal reorganization. Here we show that the activity of NF-kB markedly increases in L6 rat myogenic cells that have just initiated morphological changes by treating nocodazole, a microtubule-disrupting agent. Furthermore, the induction of NF-kB activation was closely correlated with the myoblast fusion. In addition, a variety of agents that disrupt microtubules stimulated the myoblast fusion as well as the induction of NF-kB activation. In contrast, taxol, a microtubule-stabilizing agent, suppressed the induction of NF-kB activation and inhibited spontaneous differentiation of L6 cells as well. In addition, we found that the NF-KB in the cells consists of p50/p65 heterodimers. These results support the idea that reorganization of microtubule at early stages of differentiation plays a role as a signal for NF-KB activation during myogenesis.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권3호
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• pp.193-201
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• 2006

Squamous cell carcinoma(SCC) of head and neck(SCCHN) is the sixth most common human malignant tumor. However, despite advances in prevention and treatment of SCC, the five-year survival rates for patients remain still low. To improve the outcome for patients with SCCHN, novel treatment strategies are needed. Overexpression of the epidermal growth factor(EGF) and activation of its receptor(EGFR) are associated with progressive growth of SCCHN. Vascular endothelial growth factor(VEGF) signaling molecules are related with neoangiogenesis and vascular metastasis of SCC. In this study, we determined the therapeutic effect of AEE788(Novartis Pharma AG, Basel, Switzerland), which is a dual inhibitor of EGFR/ErbB2 and VEGFR tyrosine kinases, on human oral SCC. At first, we screened the expression of EGFR, c-ErbB2(HER-2) and VEGFR-2 in a series of human oral SCC cell lines. And then we evaluated the effects of AEE788 on the phosphorylation of EGFR and VEGFR-2 in a oral SCC cell line expressing EGFR/HER-2 and VEGFR-2. We also evaluated the effects of AEE788 alone, or with paclitaxel(Taxol) on the oral SCC cell growth and apoptosis. As a result, all oral SCC cells expressed EGFR and VEGFR-2. Treatment of oral SCC cells with AEE788 led to dose-dependent inhibition of EGFR and VEGFR-2 phosphorylation, growth inhibition, and induction of apoptosis. Moreover, AEE788 sensitizes the cells to paclitaxel-mediated toxicity and apoptosis. These data mean EGFR and VEGFR-2 can be reliable targets for molecular therapy of oral SCC, and therefore warrant clinical use of EGFR/VEGFR inhibition in the treatment of patients with recurrent or metastatic oral SCC.