• Asian Pacific Journal of Cancer Prevention
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• 제15권11호
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• pp.4555-4561
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• 2014

Background: Silencing due to methylation of suppressor of cytokine signaling-3 (SOCS-3), a negative regulator gene for the JAK/STAT signaling pathway has been reported to play important roles in leukemogenesis. Imatinib mesylate is a tyrosine kinase inhibitor that specifically targets the BCR-ABL protein and induces hematological remission in patients with chronic myeloid leukemia (CML). Unfortunately, the majority of CML patients treated with imatinib develop resistance under prolonged therapy. We here investigated the methylation profile of SOCS-3 gene and its downstream effects in a BCR-ABL positive CML cells resistant to imatinib. Materials and Methods: BCR-ABL positive CML cells resistant to imatinib (K562-R) were developed by overexposure of K562 cell lines to the drug. Cytotoxicity was determined by MTS assays and $IC_{50}$ values calculated. Apoptosis assays were performed using annexin V-FITC binding assays and analyzed by flow cytometry. Methylation profiles were investigated using methylation specific PCR and sequencing analysis of SOCS-1 and SOCS-3 genes. Gene expression was assessed by quantitative real-time PCR, and protein expression and phosphorylation of STAT1, 2 and 3 were examined by Western blotting. Results: The $IC_{50}$ for imatinib on K562 was 362nM compared to 3,952nM for K562-R (p=0.001). Percentage of apoptotic cells in K562 increased upto 50% by increasing the concentration of imatinib, in contrast to only 20% in K562-R (p<0.001). A change from non-methylation of the SOCS-3 gene in K562 to complete methylation in K562-R was observed. Gene expression revealed down-regulation of both SOCS-1 and SOCS-3 genes in resistant cells. STAT3 was phosphorylated in K562-R but not K562. Conclusions: Development of cells resistant to imatinib is feasible by overexposure of the drug to the cells. Activation of STAT3 protein leads to uncontrolled cell proliferation in imatinib resistant BCR-ABL due to DNA methylation of the SOCS-3 gene. Thus SOCS-3 provides a suitable candidate for mechanisms underlying the development of imatinib resistant in CML patients.

• Toxicological Research
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• 제23권1호
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• pp.55-63
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• 2007

Diethylnitrosamine (DEN) is a nitrosamine compound that can induce a variety of liver lesions including hepatic carcinoma, forming DNA-carcinogen adducts. In the present study, microarray analyses were performed with Affymetrix Murine Genome 430A Array in order to identify the gene-expression profiles for DEN and to provide valuable information for the evaluation of potential hepatotoxicity. C57BL/6NCrj mice were orally administered once with DEN at doses of 0, 3, 7 and 20 mg/kg. Liver from each animal was removed 2, 4, 8 and 24 hrs after the administration. The histopathological analysis and serum biochemical analysis showed no significant difference in DEN-treated groups compared to control group. Conversely, the principal component analysis (PCA) profiles demonstrated that a specific normal gene expression profile in control groups differed clearly from the expression profiles of DEN-treated groups. Within groups, a little variance was found between individuals. Student's t-test on the results obtained from triplicate hybridizations was performed to identify those genes with statistically significant changes in the expression. Statistical analysis revealed that 11 genes were significantly downregulated and 28 genes were upregulated in all three animals after 2 h treatment at 20 mg/kg. The upregulated group included genes encoding Gdf15, JunD1, and Mdm2, while the genes including Sox6, Shmt2, and SIc6a6 were largely down regulated. Hierarchical clustering of gene expression also allowed the identification of functionally related clusters that encode proteins related to metabolism, and MAPK signaling pathway. Taken together, this study suggests that match with a toxicant signature can assign a putative mechanism of action to the test compound if is established a database containing response patterns to various toxic compounds.

• 한국버섯학회지
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• 제14권4호
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• pp.211-214
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• 2016

'건이'는 2012년 전남농업기술원에서 육성한 털목이 품종으로 2009년 JNM-Mi-21008과 JNM-Mi-21014의 단포자 교배를 통하여 2010년 1차 선발하였다. 2011년부터 2012년까지 특성 및 생산력 검정을 실시한 결과 갓색이 연갈색으로 우수하고 병해충에 강한 계통번호 JNMMI249를 우수균주로 선발하였고 이 계통을 2013년 '건이'라는 이름으로 품종보호출원하여 2016년 등록하였다(제109호: 2016. 6.9.). 건이의 갓은 연갈색으로 매끈하여 기호성이 높으며 씹힘성이 좋고 식이섬유 함량이 높다. 원균배양에 알맞은 배지는 MCM, 맥아, PDA배지이다. 톱밥 봉지재배시 자실체 특성으로 갓색은 연갈색이며 유효경수는 39개/0.9 kg로 많았고 갓 직경 6.9 cm, 갓 폭 8.7 cm로 갓 크기는 작은 경향이다. 배양기간은 $20^{\circ}C$에서 40~54일이며 발이 및 생육기간은 20일로 기존품종보다 다소 긴 편이며 수량은 0.9 kg 봉지당 291 g 정도이다. 또한 병해충에 강하며 2차 균사발생, 부패 등 불량환경조건에서도 우수한 경향을 보였다. 교배모본 및 대조구와의 대치선이 형성되었으며 RAPD를 이용한 유전적 다형성 검정 결과 모균주와 유의적 구별성을 보였다. 또한 식이섬유 함량이 높고 씹힘성이 우수한 특성을 갖고 있는 것으로 나타났다.

• Childhood Kidney Diseases
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• 제15권1호
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• pp.38-48
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• 2011

목 적: 국소성 분절성 사구체 경화증(Focal segmental glomerulosclerosis, 이하 FSGS)은 소아신부전의 원인 중 가장 흔한 사구체 질환이다. 일차성 FSGS의 병인은 아직 알려져 있지 않으므로, 저자들은 FSGS의 동물 모델을 대상으로 cDNA 마이크로어레이를 이용한 유전자 발현 양상 분석을 통하여 유전자 발현 수준에서의 FSGS의 질환의 특성을 밝히고자 하였다. 방 법: 사람의 일차성 FSGS와 유사한 질병경과를 보이는 동물모델인 FGS/kist 생쥐의 신피질 조직을 대조군 생쥐(FGS/kist 생쥐의 조상 strain인 RFM/kist 생쥐)와 AB 1700 mouse chip을 이용한 마이크로어레이 실험으로 비교하였다. 결 과: FGS 질병특이 유전자가 62개 추출되었다. 이들은 세포주기/사멸, 면역반응과 지질 대사/혈관 질환과 관련된 유전자들로써, 유전자간 network의 중심유전자가 면역반응(TNF, IL-6/4, IFNg)과 세포사멸 조절 유전자(TP 53), 그리고 지질대사의 중요 유전자인 PPARG이었다. 결 론: 이 연구에서 저자들은 자발적인 FSGS의 임상경과를 보이는 FGS/Kist 생쥐의 신장조직의 유전자 발현의 분석을 통하여 신장세포사멸과 면역반응에 뒤따르는 기질 섬유화, 그리고 지질 대사의 이상과 조기 혈관 질환이 FSGS의 병태생리에 기여할 것임을 다시 확인할 수 있었다. 추가적인 연구가 계속된다면 global transcriptome profiling 기법으로 병인 탐색 및 치료방법 개발 에 의미 있는 결과를 도출할 수 있을 것이다.

• Asian-Australasian Journal of Animal Sciences
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• 제27권9호
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• pp.1254-1262
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• 2014

Lymphoid enhancer binding factor 1 (LEF-1) is a member of the T-cell specific factor (TCF) family, which plays a key role in the development of breast endothelial cells. Moreover, LEF-1 gene has been identified as a candidate gene for teat number trait. In the present study, we detected two novel mutations (NC_010450.3:g. 99514A>G, 119846C>T) by DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism in exon 4 and intron 9 of LEF-1 in Guanzhong Black, Hanjiang Black, Bamei and Large White pigs. Furthermore, we analyzed the association between the genetic variations with teat number trait in these breeds. The 99514A>G mutation showed an extremely significant statistical relevance between different genotypes and teat number trait in Guanzhong (p<0.001) and Large White (p = 0.002), and significant relevance in Hanjiang (p = 0.017); the 119846C>T mutation suggested significant association in Guanzhong Black pigs (p = 0.042) and Large White pigs (p = 0.003). The individuals with "AG" or "GG" genotype displayed more teat numbers than those with "AA"; the individuals with "TC" or "CC" genotype showed more teat numbers than those with "TT". Our findings suggested that the 99514A>G and 119846C>T mutations of LEF-1 affected porcine teat number trait and could be used in breeding strategies to accelerate porcine teat number trait improvement of indigenous pigs breeds through molecular marker assisted selection.

• 한국산학기술학회논문지
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• 제15권11호
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• pp.6774-6781
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• 2014

인삼(Panax ginseng C. A. Meyer)의 주요 생리활성물질인 진세노사이드(ginsenoside) Rb1과 Rg1의 효능검증 및 작용점을 규명하고자 HaCaT 피부각질세포에서 유전체 분석(gene expression profiles)을 실시하였다. 진세노사이드 Rb1과 Rg1 각각의 처리 농도 및 시간에 따른 HaCaT 세포에 대한 세포독성은 나타나지 않았으며, $10{\mu}g/mL$의 진세노사이드 Rb1과 Rg1 각각을 6 및 24 시간 처리하여 유전체 분석 결과, 진세노사이드 Rb1과 Rg1의 24 시간 처리군에서 항노화 및 피부탄력 관련 유전자인 fibroblast growth factor (FGF2)의 활성이 증가된 것으로 나타났다. 또한 진세노사이드 Rb1의 24 시간 처리군에서는 항산화 작용점에 있는 일련의 유전자군, FANCD2, FGF2, LEPR, FAS 등의 활성을 확인하였다. 향후 확인된 항노화 및 피부탄력 관련 주요인자들의 작용 및 상관관계를 구체적으로 확인하고, 특히 진세노사이드 Rb1의 신호전달을 완성하고자 한다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권1호
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• pp.495-499
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• 2014

Estrogens are considered the major breast cancer risk factor, and the carcinogenic potential of estrogens might be attributed to DNA modification caused by derivatives formed during metabolism. $17{\beta}$-estradiol ($E_2$), the main steroidal estrogen present in women, is metabolized via two major pathways: formation of 2-hydroxyestradiol (2-OH $E_2$) and 4-hydroxyestradiol ($4-OH\;E_2$) through the action of cytochrome P450 (CYP) 1A1 and 1B1, respectively. Previous reports suggested that $2-OH\;E_2$ has putative protective effects, while $4-OH\;E_2$ is genotoxic and has potent carcinogenic activity. Thus, the ratio of $2-OH\;E_2/4-OH\;E_2$ is a critical determinant of the toxicity of $E_2$ in mammary cells. In the present study, we investigated the effects of berberine on the expression profile of the estrogen metabolizing enzymes CYP1A1 and CYP1B1 in breast cancer MCF-7 cells. Berberine treatment produced significant induction of both forms at the level of mRNA expression, but with increased doses produced 16~ to 52~fold greater induction of CYP1A1 mRNA over CYP1B1 mRNA. Furthermore, berberine dramatically increased CYP1A1 protein levels but did not influence CYP1B1 protein levels in MCF-7 cells. In conclusion, we present the first report to show that berberine may provide protection against breast cancer by altering the ratio of CYP1A1/CYP1B1, could redirect $E_2$ metabolism in a more protective pathway in breast cancer MCF-7 cells.

• Tuberculosis and Respiratory Diseases
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• 제67권1호
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• pp.8-13
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• 2009

Background: The insulin receptor substrate-1 (IRS-1) is the primary docking molecule for the insulin-like growth factor I receptor (IGF-IR), and is required for activation of the phosphatidylinositol 3'-kinase (PI3K) pathway. IRS-1 activation of the (PI3K) pathway regulates IGF-mediated survival, enhancement of cellular motility and apoptosis. Therefore, we attempted to ascertain whether IRS-1 genetic variations affect an individual's risk for non-small cell lung cancer (NSCLC). Methods: Two-hundred and eighteen subjects, either diagnosed with NSCLC or control subjects, were matched by age, gender and smoking status. Genomic DNA from each subject was amplified by PCR and analyzed according to the restriction fragment length polymorphism (RFLP) profile to detect the IRS-1 G972R polymorphism. Results: The frequencies of each polymorphic variation, in the control population, were as follows: GG=103 (94.5%) and GR=6 (5.5%); for the NSCLC subjects, the genotypic frequencies were as follows: GG=106 (97.2%) and GR=3 (2.8%). We could not demonstrate statistically significant differences in the genotypic distribution between the NSCLC and the control subjects (p=0.499, Fisher's Exact test). The relative risk of NSCLC, associated with the IRS-1 G972R polymorphic variation, was 1.028 (95% CI; 0.63~9.90). In addition, we found no differences between polymorphic variants with regard to the histological subtype of NSCLC. Conclusion: We did not observe any noteworthy differences in the frequency of the IRS-1 G972R polymorphism in NSCLC patients, compared to control subjects. These results suggest suggesting that, in our study population, the IRS-1 G972R polymorphism does may not appear to be associated with an increased risk of NSCLC.

• 대한한방소아과학회지
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• 제16권1호
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• pp.39-74
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• 2002

Recently, increased attention has been paid to the growth of the height of children and adolescents. To accelerate growth, velvet antlers are typically used in Oriental medicine. The present study investigated the effects of velvet antlers of velvet antlers on bone growth using the cell line of Human Osteosarcoma (Hos), derived from the bone-generating cells essential to bone growth. In order to give certain stress to this Hos, the medium contained 1% FBS was used for culturing for Hos cell instead of 10% in control. In this condition of which the proliferation had been significantly decreased, the ethanol extract of upper part of velvet antlers was added, As a result, the cells proliferation rate was significantly increased. Using Oligonucleotide DNA microarray, comparison and analyses were done to see what kind of specific genes would be differentially expressed. The result showed that as opposed to the control group, the stressed group indicated a decrease in the expressions of 6 kinds of genes such as, Id1, retinoid X receptor(RXRB) and 14-3-3 epsilon, etc. The velvet antler treated group, as opposed to the control group, showed a decreased in the expressions of 8 kinds of genes such as Id1, etc. and an increase in the expressions of 24 kinds of genes. The number of genes that showed differences in the velvet antler treated group compared with the stressed group was 7 the expression of 1 kind of gene was decreased, and the expressions of 6 kinds of genes were increased. Considering the mechanism by which velvet antlers affected the growth of osteoblast through reviewing the functions of these genes, the following results were attained. The constraint in the proliferation of Hos cells resulting from the medium contained 1% FBS seems to be caused by three important factors: 1) the decrease of the expression of 14-3-3 epsilon involved in the signal transduction and metabolism of growth, 2) the decrease of the expression of Id1 gene involved in the metabolism of bone formation, and 3) the decreased of expression of RXRB gene involved in the metabolism of retinoci acid. It is suggested that the improvement of the cell proliferating effects by velvet antler treatment, in stressed condition si mediated by increment of 6 genes particularly 14-3-3 epsilon, RXRB, and IGF2, with are the crucial factors for the cell growth and differentiation, metabolism of retinoic acid and osteoblast proliferation, respectively.

• 동의생리병리학회지
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• 제17권3호
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• pp.633-642
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• 2003

A pain is the symptom which defends against noxious stimulus about a human body, it is known that if the periphery of perceptive nerve were stimulated by a physical or chemical factors, the stimulation is induced by transmission to pain center in the cerebral cortex according to pain conduction tract. The treatment of pain is to decrease a stimulus that causes a pain or block off a nerve transmitting a stimulus or puts on a way to calm down pain center, but It is for adjustment of a pain to be the most representative in acupuncture among various ways to cure a pain in Oriental medicine. However, the analgesic effect of an individual response to acupuncture stimulation shows marked individual variations, so these days genetic a few approach is attempted. On this the author determined that the responding group was appointed those whose tail flick latency (TFL) responding time delayed the minimum of 30 % comparing with basal reaction time. For those whose TFL time had shorter than 30 % was grouped as a non-responding group. And then the hypothalamus of each group was dissected and RNA was further purified. After synthesizing cDNA using oligo dT primer, products were finally applied to the PCR. The results were as follows; The ratio of responding group to non-responding group was 6:4. Ach T (acetylcholinesterase T subunit), BF-I (Brain factor-I), DBH (Dopamine β-hydroxylase) and PNM (Phosphotidylethanolamine N-Methyltransferase) were revealed significantly in the responding group. Cathepsin B and Tau were revealed significantly in the non-responding group. The PCR results show that Ach T, BF-I, DBH and PNM are expressed abundantly in the responding group, where as cathepsin B and tau are abundant in the non-responding group. These results suggest that the analgesic effect on acupuncture stimulation is related to regulation of neurotransmitter as well as neurodegeration of cerebrum.