• BMB Reports
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• 제40권2호
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• pp.247-260
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• 2007

Cinnamate 4-hydroxylase (C4H) is a key enzyme of phenylpropanoid pathway, which synthesizes numerous secondary metabolites to participate in development and adaption. Two C4H isoforms, the 2192-bp BnC4H-1 and 2108-bp BnC4H-2, were cloned from oilseed rape (Brassica napus). They both have two introns and a 1518-bp open reading frame encoding a 505-amino-acid polypeptide. BnC4H-1 is 57.73 kDa with an isoelectric point of 9.11, while 57.75 kDa and 9.13 for BnC4H-2. They share only 80.6% identities on nucleotide level but 96.6% identities and 98.4% positives on protein level. Showing highest homologies to Arabidopsis thaliana C4H, they possess a conserved p450 domain and all P450-featured motifs, and are identical to typical C4Hs at substrate-recognition sites and active site residues. They are most probably associated with endoplasmic reticulum by one or both of the N- and C-terminal transmembrane helices. Phosphorylation may be a necessary post-translational modification. Their secondary structures are dominated by alpha helices and random coils. Most helices locate in the central region, while extended strands mainly distribute before and after this region. Southern blot indicated about 9 or more C4H paralogs in B. napus. In hypocotyl, cotyledon, stem, flower, bud, young- and middle-stage seed, they are co-dominantly expressed. In root and old seed, BnC4H-2 is dominant over BnC4H-1, with a reverse trend in leaf and pericarp. Paralogous C4H numbers in Brassicaceae genomes and possible roles of conserved motifs in 5' UTR and the 2nd intron are discussed.

• 대한두경부종양학회지
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• 제27권2호
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• pp.190-197
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• 2011

Background and Objective : Radiation resistance(RR) is one of main determinants of treatment outcome in oral squamous cell carcinoma(OSCC), but accurate prediction of RR is difficult. We aim to establish RR OSCC cell lines and identify genes related with RR by a measurement of altered gene expression after inducing RR. Material and Methods : OSCC cell lines, SCC15, SCC25 and QLL1, were treated with 2Gy radiation per session, and parts of them were alive in finally accumulated dosage of 60Gy through 30 times repetition of radiotherapy for inducing RR cell lines. We compared results of cDNA array and proteomics in non-radiated cell lines and RR cell lines to detect changes of gene expression. Western blot was used for the validation of results. Results : cDNA array revealed 265 commonly up-regulated genes and 268 commonly down-regulated genes in 3 RR cell lines comparing their non-radiated counterpart. Among them, 30 cancer related genes were obtained. Proteomics showed 51 commonly altered protein expressions in 3 RR cell lines and 18 cancer related proteins were obtained. Among the detected genes, we found NM23-H1 and PA2G4 were over-expressed in both cDNA array and proteomics. Western blot showed increased expression of NME1 in RR cell lines but not in PA2G4. Conclusion: We concluded that NM23-H1 may be a candidate of RR related gene and over-expression of NM23-H1 could be a biomarker to predict RR in OSCC.

• Animal cells and systems
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• 제2권4호
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• pp.483-488
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• 1998

We previously found that a potent gonadotropin-releasing hormone (GnRH) agonist, buserelin, decreases GnRH promoter activity together with GnRH mRNA level, providing evidence for an autoregulatory mechanism operating at the level of GnRH gene transcription in immortalized GT1-1 neuronal cells. To examine whether agonist-induced decrease in GnRH mRNA level requires the continuous presence of buserelin, we performed a pulse-chase experiment of buserelin treatment. Short-term exposure (15 min) of GT1-1 neuronal cells to buserelin ($10{\mu}M$) was able to decrease GnRH mRNA levels when determined 24 h later. When GT1-1 cells were treated with buserelin ( $10{\mu}M$) for 30 min and then incubated for 1, 3, 6, 12, 24, and 48 h after buserelin removal, a significant decrease in GnRH mRNA levels was observed after the 12 h incubation period. These data indicate that inhibitory signaling upon buserelin treatment may occur rapidly, but requires a long time (at least 12 h) to significantly decrease the GnRH mRNA level. To examine the possible involvement of de novo synthesis and/or mRNA stability in buserelin-induced decrease in GnRH gene expression, actinomycin D ($5{\mu}m/ml$), a potent RNA synthesis blocker, was co-treated with buserelin. Actinomycin D alone failed to alter basal GnRH mRNA Revel, but blocked the buserelin-induced decrease in GnRH mRNA level at 12 h of post-treatment. These data suggest that buserelin may exert its inhibitory action by altering the stability of GnRH mRNA. Moreover, a polvsomal RNA separation by sucrose gradient centrifugation demonstrated that buserelin decreased the translational efficiency of the transcribed GnRH mRNA. Taken together, these results clearly indicate that GnRH agonist buserelin acts as an inhibitory signal at multiple levels such as transcription mRNA stability, and translation.

• Asian Pacific Journal of Cancer Prevention
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• 제15권24호
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• pp.10697-10703
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• 2015

The laryngeal squamous cell carcinoma (LSCC) is one of the most common malignant tumors occurring in the head and neck. Tumor necrosis factor related apoptosis induce ligand (TRAIL) and TRAIL-receptors (DR4, DR5, DcR1, DcR2) are known as important members of TRAIL-mediated biochemical signaling pathway. Associations between polymorphisms in these genes and clinicopathological characteristics of human laryngeal carcinoma are not well defined. This study therefore aimed to investigate a possible relationship among the TRAIL and TRAIL-DR4 polymorphisms and sTRAIL levels in the risk or progression of LSCC. A total of 99 patients with laryngeal cancer and 120 healthy subjects were enrolled in the study. DR4 C626G and TRAIL 1595 C/T genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and sTRAIL levels were measured by ELISA. There were significant differences in the distribution of DR4 C626G genotypes and frequencies of the alleles between laryngeal cancer patients and controls (p<0.001) but not in TRAIL 1595 C/T. We found the increased frequency of the DR4 C626G homozygote CC genotype in patients than in controls (p<0.001). Haplotype analysis revealed that there was also a statistically significant relationship between TRAIL and TRAIL-DR4 polymorphisms and laryngeal cancer. Serum sTRAIL levels in the laryngeal patients with CC genotype who had advanced tumour stage were lower than those of patients with early tumor stage (p=0.014). Our findings suggest that DR4 C626G genotypes and sTRAIL levels might be associated with progression of laryngeal cancer in the Turkish population.

• Journal of Gastric Cancer
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• 제21권4호
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• pp.368-378
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• 2021

Purpose: When patients with early gastric cancer (EGC) undergo non-curative endoscopic submucosal dissection requiring gastrectomy (NC-ESD-RG), additional medical resources and expenses are required for surgery. To reduce this burden, predictive model for NC-ESD-RG is required. Materials and Methods: Data from 2,997 patients undergoing ESD for 3,127 forceps biopsy-proven differentiated-type EGCs (2,345 and 782 in training and validation sets, respectively) were reviewed. Using the training set, the logistic stepwise regression analysis determined the independent predictors of NC-ESD-RG (NC-ESD other than cases with lateral resection margin involvement or piecemeal resection as the only non-curative factor). Using these predictors, a risk-scoring system for predicting NC-ESD-RG was developed. Performance of the predictive model was examined internally with the validation set. Results: Rate of NC-ESD-RG was 17.3%. Independent pre-ESD predictors for NC-ESD-RG included moderately differentiated or papillary EGC, large tumor size, proximal tumor location, lesion at greater curvature, elevated or depressed morphology, and presence of ulcers. A risk-score was assigned to each predictor of NC-ESD-RG. The area under the receiver operating characteristic curve for predicting NC-ESD-RG was 0.672 in both training and validation sets. A risk-score of 5 points was the optimal cut-off value for predicting NC-ESD-RG, and the overall accuracy was 72.7%. As the total risk score increased, the predicted risk for NC-ESD-RG increased from 3.8% to 72.6%. Conclusions: We developed and validated a risk-scoring system for predicting NC-ESD-RG based on pre-ESD variables. Our risk-scoring system can facilitate informed consent and decision-making for preoperative treatment selection between ESD and surgery in patients with EGC.

• 한국환경보건학회지
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• 제43권4호
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• pp.334-348
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• 2017

Objectives: We investigated the impacts of indoor plants on indoor air quality and occupational health, focusing on allergic rhinconjunctivitis and stress among employees in new office buildings. Methods: A total of 34 employees working at new public office buildings were enrolled as subjects (n=17, with indoor plants) and as a control (n=17) group. Before and after introducing indoor plants for three months, indoor air quality measurements including VOCs and aldehydes and questionnaires on sick building syndrome, AR symptoms (ARIA based), stress (DASS 42, KOSS, and SACL), and indoor characteristics were performed and statistically analysed. Results: Among the 34 enrolled subjects, 19 were included in the probable AR subject group (subjects with indoor plants, n=8, control n=11) and completed all questionnaires. Statistical analyses were done for total, AR subject groups, and controls. As a result, it was confirmed that major indoor air pollutants decreased after the introduction of indoor plants (p<0.5). Among major symptoms of allergic rhinoconjunctivitis, watery rhinorrhea, nasal stuffiness, and nasal itching indexes decreased (p<0.5, respectively). A decrease was noted in some areas of work-related stress indexes (mainly KOSS) among the subject group (total and AR) and a decrease of indoor environmental attractiveness among the control group (total and AR) (p<0.5, for all). Conclusions: Indoor plants may help reduce indoor air pollutants and decrease AR symptoms and work-related stress of employees in newly built office buildings. Various further follow-up studies on the mechanism of environmental, physical, and emotional influences and utilization of indoor plants in association with allergic diseases will be needed.

• Asian Pacific Journal of Cancer Prevention
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• 제17권8호
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• pp.4059-4062
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• 2016

Frequencies of polymorphisms of genes BRCA1 and ТР53 in breast cancer (BC) patients with a BC family history and radiation history were assessed and compared in the Semey region of Kazakhstan. The study included 60 women directly irradiated by the activities of the Semipalatinsk test site with a calculated effective equivalent dose of 500 mSv and their first generation descendants (group BC+Her+Exp); 65 women with family BC and absence of radiological history - the effective equivalent dose due to anthropogenic sources not exceeding 50 mSv (group BC+Her-Exp). The comparison group consisted of 65 women patients with breast cancer without family and radiological history (BC-Her-Exp). The control group comprised 60 women without breast cancer and without family and radiological history (nonBC). We carried out the genotyping of the polymorphisms c.2311T>C, c.4308T>C and 5382insC of the BRCA1 gene and rs1042522 of the ТР53 gene. The frequency of the polymorphism c.2311T>C was significantly higher in patients of the group BC+Her+Exp than in healthy women, and of the polymorphism 5382insC in BC+Her+Exp compared to all other groups. The frequency of the rs1042522 polymorphism of ТР53 was significantly higher in all groups of patients with breast cancer compared with the control group. Differences between groups of women with breast cancer were significant only in BC+Her+Exp vs. BC+Her-Exp. Combinations of polymorphisms of the genes BRCA1 and TP53 predominated in women with a family and radiological history.

• 한국작물학회지
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• 제56권1호
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• pp.72-79
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• 2011

본 연구는 간이 수경재배법을 이용하여 보리의 알루미늄 스트레스 내성과 민감성 품종을 간편하고 빠르게 screen하는 방법을 소개하고, 선별된 품종간의 뿌리의 생장, 뿌리 조직의 염색, 알루미늄 함량, 원형질막의 $H^+$-ATPase의 발현 변화를 조사하여 분석하였다. l. 보리 65가지 품종을 간이 수경재배법을 이용하여 $20{\mu}M$ 알루미늄을 24시간 처리 후 뿌리생장의 차이로 내성 세 품종(자예2, 자예6, 모치무기)과 민감성 세 품종(흰쌀, 올쌀, 품2)을 선별하였다. 2. 알루미늄에 내성 품종은 알루미늄 처리 농도(0, 5, 10, $20{\mu}M$)에 따라 뿌리 생장 감소폭이 적었으나, 민감성 세 품종은 상대적으로 낮은 $5{\mu}M$ 농도에서부터 80%의 생장이 억제되었다. 3. 내성인 자예2와 민감성인 품2의 알루미늄 처리 후, 농도별(0, 5, 10, $20{\mu}M$), 시간별(3, 6, 12, 24시간)로 0.2% hematoxylin으로 염색 시 주로 apex에 3시간 이후부터 염색되었으며, 민감성 품2가 내성인 자예2에 비해 농도와 시간에 따라 그 피해 정도가 매우 심각하였다. 4. $20{\mu}M$로 24 시간 처리된 뿌리 apex(10 mm)의 알루미늄 함량을 측정한 결과, 내성인 자예2는 주당 47.1 nmol의 함량을 보여 주었으나, 민감성인 품2는 주당 64.9 nmol의 높은 함량을 보여 주었다. 5. 24시간 동안 $20{\mu}M$ 알루미늄을 처리한 뿌리 원형질막 $H^+$-ATPase 발현을 western blotting을 통해 분석한 결과, 내성인 자예2는 차이가 없었으나, 민감성 품2는 현저히 억제되었다. 이로 보아 원형질막 $H^+$-ATPase가 알루미늄의 내성 기작에 관여하는 것으로 보인다. 6. 본 연구를 통해 간이 수경재배와 hematoxylin을 이용한 염색으로 간단하고 빠르게 보리의 알루미늄 내성과 민감성 품종의 screening을 할 수 있었고, 보리뿐 아니라 쌀, 밀 등의 다른 종자에도 적용할 수 있을 것이다.

• Journal of Ginseng Research
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• 제41권3호
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• pp.247-256
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• 2017

Background: KG-135, a standardized formulation enriched with Rk1, Rg3, and Rg5 ginsenosides, has been shown to inhibit various types of cancer cells; however, the underlying mechanisms are not fully understood. In this study, we explored its effects in A549 human lung cancer cells to investigate the induction of Forkhead Class box O3a (FOXO3a) and autophagy. Methods: Cell viability was determined by sulforhodamine B staining. Apoptosis and cell cycle distribution were analyzed using flow cytometry. The changes of protein levels were determined using Western blot analysis. Autophagy induction was monitored by the formation of acidic vesicular organelles stained with acridine orange. Results: KG-135 effectively arrested the cells in G1 phase with limited apoptosis. Accordingly, a decrease of cyclin-dependent kinase-4, cyclin-dependent kinase-6, cyclin D1, and phospho-retinoblastoma protein, and an increase of p27 and p18 proteins were observed. Intriguingly, KG-135 increased the tumor suppressor FOXO3a and induced the accumulation of autophagy hallmark LC3-II and acidic vesicular organelles without an increase of the upstream marker Beclin-1. Unconventionally, the autophagy adaptor protein p62 (sequestosome 1) was increased rather than decreased. Blockade of autophagy by hydroxychloroquine dramatically potentiated KG-135-induced FOXO3a and its downstream (FasL) ligand accompanied by the cleavage of caspase-8. Meanwhile, the decrease of Bcl-2 and survivin, as well as the cleavage of caspase-9, were also drastically enhanced, resulting in massive apoptosis. Conclusion: Besides arresting the cells in G1 phase, KG-135 increased FOXO3a and induced an unconventional autophagy in A549 cells. Both the KG-135-activated extrinsic FOXO3a/FasL/caspase-8 and intrinsic caspase-9 apoptotic pathways were potentiated by blockade of autophagy. Combination of KG-135 and autophagy inhibitor may be a novel strategy as an integrative treatment for cancers.

• 지질공학
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• 제19권3호
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• pp.331-344
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• 2009

1990년 이후 우리나라의 자연사면에서 발생한 10개 대규모 산사태지역의 강우조건, 지질특성 및 기하형상에 대한 기존 자료를 재조명하였다. 이들 중 화강암 지역이 5곳, 화강암과 변성암류가 같이 분포하는 지역이 4곳이며, 나머지 1곳은 반려암이 분포한다. 총 3,435개의 산사태 중 1,992개는 화강암 분포지에서, 824개는 변성암 지역에서, 15개는 반려암에서, 그리고 나머지 604개의 산사태는 2개 지역에 부분적으로 분포하는 화산암, 퇴적암 등에서 발생하였다. 연구지역의 산사태는 강우기간 보다는 강우강도에 의하여 좌우되었다. 산사태지역의 강우자료를 Caine과 Olivier의 경험식에 적용하면 모두 Caine의 기준치(threshold)와 Olivier의 산사태반응지수(coefficient of final response)를 초과하여, 모든 지역의 강우조건이 산사태 발생조건을 충족하고 있다. 산사태의 형태와 발생 메카니즘은 기반암의 풍화특성에 따라 차이를 보이고 있다. 화강암 지역은 노두가 거의 없고 풍화심도가 고르게 분포하여 소규모의 슬라이드가 우세하다. 반면 변성암 지역은 지형이 다소 험준하고 노두가 능선과 산록에 발달하여 풍화노두가 토층과 함께 파괴, 유동하는 비교적 큰 규모의 토석류가 우세하다.