• BMB Reports
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• 제34권4호
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• pp.365-370
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• 2001

We constructed a comparative expression system in order to produce recombinant human ferritin homo- and heteropolymers in Escherichia coli. Human ferritin H-(hfH) and L-chain (hfL) genes were expressed without amino acid changes under the control of a tac promoter. Ferritin heteropolymers of varying subunit composition were also produced by combining two different expression systems, a bicistronic expression system and a coplasmid expression system. As a result, recombinant H-chain ferritin and ferritin heteropolymers were catalytically active in forming iron core in vivo. In particular, the ferritin heteropolymer that is composed of 7% H-subunit and 93% L-subunit was capable of forming an iron core of the protein, while the L-chain ferritin homopolymer was inactive in vivo. This result indicates that the two H-subunits (i.e., 7% H-subunit content) are important to keep ferritin active in the cells. In addition, human ferritins were identified as the major iron binding proteins in the transformed cells. Also, the amount of iron bound to the recombinant ferritins was proportional to the H-subunit content in ferritin heteropolymers in vivo.

• Asian Pacific Journal of Cancer Prevention
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• 제15권24호
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• pp.10917-10922
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• 2015

Background: This study was conducted to determine DEPDC1 expression in hepatocelluar carcinomas (HCCs) and to reveal its potential role in diagnosis and prognosis of affected patients. Materials and Methods: DEPDC1 expression at the mRNA level was detected by quantitative real-time PCR (qRT-PCR) in 205 cases of HCC and paired adjacent normal liver tissues, and by semi-quantitative RT-PCR in 20 cases. Survival curves were obtained by using Kaplan-Meier method and Log-rank test. Independent predictors associated with regard to disease free survival (DFS) and overall survival (OS) were identified using the Cox proportional hazard model. Results: High DEPDC1 mRNA levels were detected in 144 out of 205 cases (70.24%) of HCC, significantly associated with clinicopathological parameters, including tumor size (${\geq}4cm$), alpha-fetoprotein (${\geq}100ng/ml$), B-C of BCLC stage and recurrence. Kaplan-Meier survival analysis revealed that HCC patients with high DEPDC1 expression had poor OS and DFS. Multivariate analysis demonstrated that high DEPDC1 expression was an independent predictor for OS (HR=1.651; 95% 95%CI, 1.041-2.617; p=0.033) and DFS (HR=1.583; 95%CI, 1.01-2.483; p=0.045). Conclusions: Our results indicate DEPDC1 might be a novel diagnostic marker and an independent prognostic predictor for HCC patients.

• 대한한방부인과학회지
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• 제19권2호
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• pp.214-225
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• 2006

Purpose : This study was aimed to investigate the inhibitory effect of Corydalis Tuber on the proliferation of human uterine leiomyoma cell and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of suvival cells treated with indicated concentration of Corydalis Tuber and investigated cell viability by MTS assay. Furthermore, flow cytometric analyis were used to dissect between necrosis and apoptosis related with cell cycle and then we observed the differential gene expression by western blot analysis. Results : 1) The inhibitory effect on the proliferation of uterine leiomyoma cell treated with Corydalis Tuber was increased in a concentration and time proportional. 2) The result of flow cytometry analysis, subG1 phase arrest related cell apoptosis was not investigated in uterine leiomyoma cell treated Corydalis Tuber but showed G2/M phase prolongation. 3) The gene expression of p27, p21 related cell cycle was increased according to increasing concentration, but p53 was not exchanged. 4) The dephosphorylation of pRb gene were increased dependent on treatment concentration and pro-caspase 3, CDK4 were not exchanged. Conclusion : This study showed that Corydalis Tuber have the inhibitory effect on the proliferation of human uterine leiomyoma cell but the effect was thoughted no relationship with apoptosis. The inhibitory effect was suggested that dephosphorylation of pRb gene induced with increasing p21, p27 prolonged cell division in G2/M phase.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1563-1566
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• 2014

Background: Esophageal squamous cell carcinoma (ESCC) accounts for most esophageal cancer in Asia, and is the sixth common cause of cancer-related deaths worldwide. Previous studies indicated HOXB7 is overexpressed in ESCC tissues, but data on prognostic value are limited. Methods: A total of 76 advanced ESCC cases were investigated. Immunohistochemistry (IHC) was used to detect the expression levels of HOXB7 and Kaplan-Meier curves and Cox regression models to determine prognostic significance. Stratified analysis was also performed according to lymph node (LN) status. Results: Kaplan-Meier curve analysis indicated that HOXB7 positive patients had significantly shorter overall survival (OS) than HOXB7 negative patients. Multivariate analysis using the Cox proportional hazards model indicated only TNM stage and HOXB7 expression to be independent predictors of overall survival of advanced ESCC patients. HOXB7 indicated poor OS in both lymph node negative (LN-) and lymph node positive (LN+) patients. Conclusion: HOXB7 predicts poor prognosis of advanced ESCC patients and can be applied as an independent prognostic predictor.

• Asian Pacific Journal of Cancer Prevention
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• 제15권8호
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• pp.3613-3618
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• 2014

Stathmin, also called oncoprotein 18, is a founding member of the family of microtubule-destabilizing proteins that play a critical role in the regulation of mitosis. At the same time stathmin has been recognized as one of responsible factors in cancer cells. The aim of this study was to assess stathmin status, its correlations with clinicopathological parameters and its role as a progosnostic marker in EC patients. The protein and mRNA levels of stathmin were examined byimmunohistochemistry (IHC) and in situ hybridization in 100EC tissues and adjacent noncancerous tissues. mRNA and protein expression of stathmin in three EC cell lines(EC9706, ECa109, EC1 commonly used in research) were also analyzed using immunocytochemistry, western blot and in situ hybridization. The prognostic value of Stathmin expression within the tumor tissues were assessed by Cox regression and Kaplan-Meier analysis. We showed that stathmin expression was significantly higher in EC tissues than in adjacent noncancerous tissues. High stathmin immunostaining score in the EC was positively correlated with tumor differentiation, Tumor invasion, Lymph node metastases, and TNM stage. In addition, we demonstrated that three EC cell lines examined, were constitutively expressing a high level of stathmin. Of those, EC-1 showed the strongest mRNA and protein expression for the stathmin analyzed. Kaplan-Meier analysis showed that significantly longer 5-year survival rate was seen in EC patients with high Stathmin expression, compared to those with low expression of Stathmin expression. Furthermore, multivariate Cox proportional hazard analyses revealed that Stathmin was an independent factors affecting the overall survival probability. In conclusion, our data provide a basis for the concept that stathmin might be associated with EC development and progression. High levels of Stathmin expression in the tumor tissues may be a good prognostic marker for patients with EC.

• 한국수학교육학회지시리즈C:초등수학교육
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• 제21권4호
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• pp.445-459
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• 2018

본 연구는 시각적 표현의 활용이 비례식과 비례배분 학습에 미치는 영향을 탐구하고자 수행하였다. 이를 위해 시각적 표현에 집중하여 싱가포르 교과서와 한국 교과서의 '비례식과 비례배분'단원을 분석하고, 분석 결과를 바탕으로 한국 교과서를 시각적 표현을 중심으로 재구성하였으며, 재구성한 교재를 활용한 집단과 기존의 한국 교과서를 활용한 집단의 성취도 차이를 분석하였다. 먼저 두 교과서의 분석에서 한국 교과서는 총 85면에 6종 38개의 시각적 표현이 제시되어 있었고, 싱가포르 교과서에는 127면에 8종 152개의 시각적 표현이 활용되고 있었다. 이 결과를 바탕으로 한국 교과서를 재구성하였는데, 재구성한 단원에는 14면에 8종, 49개의 시각적 표현이 포함되었다. 마지막으로 재구성한 교재를 활용한 집단과 기존 교과서를 활용한 집단 간의 학습의 차이를 알아보기 위한 평가 후 결과를 독립표본 t-검정으로 분석한 결과 두 집단 간의 평균에 유의미한 차이를 확인할 수 있었다. 그룹간 성취 변화를 비교하고자 Mann-Whiteny 검사를 실시하였고, 상 중 하 수준별 집단 중 중위집단에서 가장 많은 효과가 있는 것으로 드러났다. 본 연구는 교과서의 시각적 표현을 분석하고 이를 통해 교재를 재구성, 현장에 투입함으로써 시각적 표현이 비례식과 비례배분의 학습에 긍정적 영향을 준다는 것을 밝혀냈다는 점에서 의의를 찾을 수 있다.

• 식물조직배양학회지
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• 제22권1호
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• pp.19-23
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• 1995

당근의 현탁배양 캘러스 세포가 고온에 노출되었을때 다른 식물에서 알려진 것과 유사한 열충격반응을 나타낸다. 이는 기존의 유전자의 발현을 억제하고 새로운 열충격단백질을 생산하기 위한 열충격유전자의 발현을 촉진한다. 이러한 열충격유전자의 고온조건에서의 발현은 주로 전사과정에서 이루어지는 것으로 알려져 있다. 그러나 몇가지의 경우에서 이들 열충격 유전자의 발현이 번역과정에서 조절되는 것으로 보고된 바 있다. 본 고에서는 열충격 과정과는 무관한 토끼의 적혈구로 만들어진 시험관 내 번역시스템과 2차원단백질분리 시스템을 이용하여 생성된 단백질의 양을 통해, 번역에 사용된 mRNA의 양을 추정하였다. 이를 생체내에서 같은 조건에서 만들어진 단백질의 양과 비교할 때에 당근세포내의 특정 mRNA의 양과 해당 열충격단백질의 양이 $ 30^{\circ}C$에서 불일치함을 확인하였다. 이를 통해 당근의 캘러스 세포가 열충격 반응을 나타내는 가장 낮은 온도인 $30^{\circ}C$에서 mRNA의 양과는 다소 무관하게 해당 열충격단백질을 번역을 촉진하는 과정이 있음을 추정할 수 있었다.

• Journal of Plant Biotechnology
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• 제4권4호
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• pp.143-150
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• 2002

A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7681-7684
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• 2013

Background: Human leukocyte antigen (HLA)-G-positive gastric cancers are associated with poor survival, but links with tumor escape mechanisms remain to be determined. Materials and Methods: We used immunohistochemistry to investigate HLA-G expression, tumor infiltrating CD8+ T lymphocytes, and Treg cells in 52 gastric cancer patients. Results: There were 29 cancer-related deaths during the follow-up period. Kaplan-Meier analysis indicated that patients with HLA-G-positive (n=16) primary tumors had a significantly poorer prognosis than patients with HLA-G-negative tumors (n=36, p=0.008). The median survival time was 14 months and 47 months, respectively. Patients with high numbers of Tregs and low numbers of CD8+T lymphocytes in the primary tumor had a poorer prognosis than those with low numbers of Tregs and high numbers of CD8+T lymphocytes (p=0.034, p=0.043). Multivariate Cox proportional hazard regression analysis showed that HLA-G expression (hazard ratio: 2.662; 95% confidence interval: 1.242-5.723; p=0.012) and stage (hazard ratio: 2.012;95% confidence interval: 1.112-3.715; p=0.041) were independent unfavorable factors for patient survival. Conclusions: We found a significant positive correlation between HLA-G expression and the number of tumor infiltrating Tregs (p=0.01) and a negative correlation with the number of CD8+T lymphocytes (p=0.041). HLA-G may protect gastric cancer cells from cytolysis by inducing Foxp3+Treg lymphocytes and suppressing CD8+T lymphocytes.

• Journal of Microbiology and Biotechnology
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• 제20권2호
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• pp.356-362
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• 2010

Ceramide is important not only for the maintenance of the barrier function of the skin but also for the water-binding capacity of the stratum corneum. Although the exact role of ceramide in the human skin is not fully understood, ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries. Compared with other microorganisms, yeast is more suitable for the production of ceramide because yeast grows fast and is non-toxic. However, production of ceramide from yeast has not been widely studied and most work in this area has been carried out using Saccharomyces cerevisiae. Regulating the genes that are involved in sphingolipid synthesis is necessary to increase ceramide production. In this study, we investigated the effect of the genes involved in the synthesis of ceramide, lcb1, lcb2, tsc10, lac1, lag1, and sur2, on ceramide production levels. The genes were cloned into pYES2 high copy number vectors. S. cerevisiae was cultivated on YPDG medium at $30^{\circ}C$. Ceramide was purified from the cell extracts by solvent extraction and the ceramide content was analyzed by HPLC using ELSD. The maximum production of ceramide (9.8 mg ceramide/g cell) was obtained when the tsc10 gene was amplified by the pYES2 vector. Real-time RT-PCR analysis showed that the increase in ceramide content was proportional to the increase in the tsc10 gene expression level, which was 4.56 times higher than that of the control strain.