• Molecular & Cellular Toxicology
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• 제4권3호
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• pp.199-204
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• 2008

The bio-molecules involved in defense mechanisms can be used as efficient biomarkers for physiological changes in organisms caused by both of internal and external stress. Thus, the expression level of genes which encoding such molecules serve as critical 'early warning system' for environmental assessment as well as health diagnosis of biological organisms. In this study, Cytochrome P450, Heat shock protein 90, Ubiquitin and ${\beta}$-actin gene were isolated for the first time from a red alga Gracilaria textorii. The quantitative differential gene expression analyses of three genes, GteCYP1A, GteHsp90 and Gte-UB, were carried out in G. textorii sporophytes collected from two different localities, polluted Sujeong (Masan, Korea) and potentially unpolluted Danggeum (Daemaemuldo Is., Korea). The transcripts of all three tested genes were highly expressed in the Sujeong population. The results suggest: 1) the Sujeong site was more polluted than the Danggeum site; 2) G. textorii could be applicable to marine environment monitoring in coastal regions.

• BMB Reports
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• 제45권6호
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• pp.331-336
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• 2012

The retinoid-related orphan nuclear receptor gamma ($ROR{\gamma}$) plays critical roles in regulation of development, immunity and metabolism. As transcription factor usually forms a protein complex to function, thus capturing and dissecting of the $ROR{\gamma}$ protein complex will be helpful for exploring the mechanisms underlying those functions. After construction of the recombinant tandem affinity purification (TAP) plasmid, pMSCVpuro $ROR{\gamma}$-CTAP(SG), the nuclear localization of $ROR{\gamma}$-CTAP(SG) fusion protein was verified. Following isolation of $ROR{\gamma}$ protein complex by TAP strategy, seven candidate interacting proteins were identified. Finally, the heat shock protein 90 (HSP90) and receptor-interacting protein 140 (RIP140) were confirmed to interplay with $ROR{\gamma}$ by co-immunoprecipitation. Interference of HSP90 or/and RIP140 genes resulted in dramatically decreased expression of CYP2C8 gene, the $ROR{\gamma}$ target gene. Data from this study demonstrate that HSP90 and RIP140 proteins interact with $ROR{\gamma}$ protein in a complex format and function as co-activators in the $ROR{\gamma}$-mediated regulatory processes of HepG2 cells.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 1997년도 제34차 추계 학술대회
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• pp.50.2-51
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• 1997

• Asian-Australasian Journal of Animal Sciences
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• 제23권4호
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• pp.437-443
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• 2010

To be economically profitable, the poultry industry demands an increase in stocking density, which could adversely affect chicken welfare. The current study was performed to investigate the effect of stocking density on stress-related, heat shock protein genes (HSP70 and HSP90), 3-hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) gene and telomere length in broiler chickens. Seven-day-old broiler chickens were housed at High (0.0578 $m^2$/bird), Standard (0.077 $m^2$/bird) and Low (0.116 $m^2$/bird) stocking densities with 8 replicates each until 35 d of age. The growth performance, such as body weight gain and average daily feed intake, was found to be significantly (p<0.05) higher in the Low density group, but these parameters did not show any difference between the High and Standard groups. Other growth performance, such as feed conversion ratio and final feed intake, showed no difference among the treated groups. The expression levels of HSP70 and HMGCR were found to be elevated with the increase of stocking density. The expression level of these genes was significantly (p<0.05) higher in the High density stocked group compared with the other groups, whereas the expression levels were not significantly different between the Low and Standard groups. The expression levels of HSP90 did not show any significant changes among the treated groups. The telomeric length of the birds housed in High density was reduced significantly (p<0.05) when compared to that of the birds in Low density. These results clearly indicate that birds stocked at high density show physiological adaptive changes indicative of stress at gene transcriptional and telomere levels.

• Animal Bioscience
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• 제36권12호
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• pp.1796-1805
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• 2023

Objective: This study aims to identify heat shock protein70-2 (HSP70-2) and protamine-1 (PRM1) mRNA and protein in Madura bull sperm and demonstrate their relation as bull fertility biomarkers. Methods: The Madura bull fertility rates were grouped based on the percentage of first service conception rate (%FSCR) as high fertility (HF) (79.04%; n = 4), and low fertility (LF) (65.84%; n = 4). mRNA of HSP70-2 and PRM1 with peptidylprolyl isomerase A (PPIA) as a housekeeping gene were determined by quantitative real-time polymerase chain reaction, while enzyme-linked immunoassay was used to measure protein abundance. In the post-thawed semen samples, sperm motility, viability, acrosome integrity, and sperm DNA fragmentation index were analyzed. Data analysis was performed on the measured parameters of semen quality, relative mRNA expression, and protein abundance of HSP70-2 and PRM1, among the bulls with various fertility levels (HF and LF) in a one-way analysis of variance analysis. The Pearson correlation was used to analyze the relationship between semen quality, mRNA, proteins, and fertility rate. Results: Relative mRNA expression and protein abundance of HSP70-2 and PRM1 were detected and were found to be highly expressed in bulls with HF (p<0.05) and were associated with several parameters of semen quality. Conclusion: HSP70-2 and PRM1 mRNA and protein molecules have great potential to serve as molecular markers for determining bull fertility.

• Journal of Gastric Cancer
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• 제4권3호
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• pp.169-175
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• 2004

Purpose: The tumor suppressor gene p53 has been shown to be a factor in the carcinogenesis or progression of gastric cancer. The mutant p53 has been reported to cause a higher risk of lymph-node metastasis. Futhermore, mutation of the p53 has been linked to a poor prognosis for gastric cancer. The heat shock protein-27 (HSP27), a stress protein, has also been reported to be a poor prognostic factor in ovarian and breast cancers. However, in gastric-cancer patients, controversies exist as to its influence on the prognosis. In the present study, we used an immunohistochemical stain to observe the effects of p53 and HSP27 on the clinicopathological factors and on the prognosis for gastric-cancer patients. Materials and Methods: To evaluate the significance of p53 and HSP27 in gastric cancer patients, we analyzed 212 cases of gastric cancer (stage I.IV). Tissue samples of 212 patients were stained immunohistochemically for the mutant p53 protein and for HSP27. The correlations between protein expression and the clinicopathological factors were investigated. Results: The overall expression rates for p53 and HSP27 were $36.9\%\;and\;27.8\%$, respectively. p53 and HSP27 were correlated to each other because the HSP27 expression rate was higher in the p53-positive group (P=0.046). Statistically, the p53 and the HSP27 expression rates were significantly increased in the case of tumor invasiveness, lymphatic metastasis and vessel involvement. Therefore, they play a role in cancer progression. The 5-year survival rates of the p53-positive and the p53-negative groups were $62.8\%\;and\;60.1\%$, respectively (P=0.793) while the 5-year survival rates for the HSP27-positive and HSP27-negative groups were $54.2\%\;and\;63.1\%$, respectively (P=0.090). Conclusion: p53 and HSP27 were correlated to each other in our immunohistochemical study of gastric carcinomas and they were not independent prognostic factors in gastric- cancer patients. However, further studies are needed to determine their prognostic values for gastric-cancer patients.

• BMB Reports
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• 제48권3호
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• pp.178-183
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• 2015

Dendritic cells play an important role in determining whether na${\ddot{i}}$ve T cells mature into either Th1 or Th2 cells. We determined whether heat-shock protein X (HspX) purified from Mycobacterium tuberculosis regulates the Th1/Th2 immune response in an ovalbumin (OVA)-induced murine model of asthma. HspX increased interferon-gamma, IL-17A, -12 and transforming growth factor (TGF)-${\beta}$ production and T-bet gene expression but reduced IL-13 production and GATA-3 gene expression. HspX also inhibited asthmatic reactions as demonstrated by an increase in the number of eosinophils in bronchoalveolar lavage fluid, inflammatory cell infiltration in lung tissues, airway luminal narrowing, and airway hyper-responsiveness. Furthermore, HspX enhanced OVA-induced decrease of regulatory T cells in the mediastinal lymph nodes. This study provides evidence that HspX plays critical roles in the amelioration of asthmatic inflammation in mice. These findings provide new insights into the immunotherapeutic role of HspX with respect to its effects on a murine model of asthma.

• 생명과학회지
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• 제26권9호
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• pp.991-998
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• 2016

NF-κB는 anti-apoptotic gene을 유도하는 전사인자로서 대부분의 세포의 생존에 필요하다. 그러나 NF-κB가 많은 종류의 암세포에서 지속적으로 과다 활성화됨이 알려지면서 NF-κB의 활성억제가 암의 예방과 치료에 유효하다는 점이 알려지게 되었다. 한편, Hsp70가 NF-κB의 활성을 조절한다는 사실이 알려지면서 Hsp70를 이용한 암예방과 치료가 주목받게 되었으나 아직 Hsp70에 의한 NF-κB의 활성조절기전은 명확하지 않다. 본 연구에서는 Hsp70에 의한 NF-κB의 활성조절과정에서 IKK complex의 구성성분인 IKKγ의 역할을 검토하였다. IKKγ의 wild type과 deletion mutants를 이용하여 Hsp70와 관련된 NF-κB의 활성조절을 연구한 결과 Hsp70는 NF-κB의 활성화를 억제하였으며, 이러한 억제효과는 IKKγ가 과발현되었을 때 더욱 증가하였다. 또한 IKKγ의 N-말단의 IKKβ 결합부위와 C-말단의 Leucine zipper 및 Zinc finger부위는 Hsp70와 연관된 NF-κB억제작용에 필요하지 않는 것으로 나타났으며, Hsp70와 IKKγ에 의한 NF-κB의 활성억제는 IκBα의 인산화와 분해를 저해함에 의해 일어나는 것으로 나타났다. 또한 RAW264.7 macrophage세포에서 LPS에 의한 COX-2의 발현유도는 Hsp70와 IKKγ가 동시에 발현 되었을 때 가장 효과적으로 억제되었다. 이상의 결과로부터 Hsp70에 의한 NF-κB의 활성억제작용은 IKKγ에 의해 상승됨을 알 수 있었으며, Hsp70와 IKKγ를 적절히 이용하면 NF-κB의 과다활성에 의해 발생하는 각종 질병의 예방과 치료에 도움을 줄 수 있을 것으로 기대된다.

• 한국패류학회지
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• 제29권3호
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• pp.181-187
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• 2013

In order to investigate environmental stress inducible genes in abalone, we analyzed differentially expressed transcripts from a Pacific abalone, Haliotis discus hannai, after exposure to heat-, cold- or hyposalinity-shock by suppression subtractive hybridization (SSH) method. 1,074 unique sequences from SSH libraries were composed to 115 clusters and 986 singletons, the overall redundancy of the library was 16.3%. From the BLAST search, of the 1,316 ESTs, 998 ESTs (75.8%) were identified as known genes, but 318 clones (24.2%) did not match to any previously described genes. From the comparison results of ESTs pattern of three SSH cDNA libraries, the most abundant EST was different in each SSH library: small heat shock protein p26 (sHSP26) in heat-shock, trypsinogen 2 in cold-shock, and actin in hyposalinity SSH cDNA library. Based on sequence similarities, several response-to-stress genes such as heat shock proteins (HSPs) were identified commonly from the abalone SSH libraries. HSP70 gene was induced by environmental stress regardless of temperature-shock or salinity-stress, while the increase of sHSP26 mRNA expression was not detected in cold-shock but in heat-shock condition. These results suggest that the suppression subtractive hybridization method is an efficient way to isolate differentially expressed gene from the invertebrate environmental stress-response transcriptome.

• 한국해양생명과학회지
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• 제3권2호
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• pp.51-58
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• 2018

수온의 변화는 어류의 거의 모든 생리학적 부분에 영향을 미친다. 기후 변화로 인한 수온의 상승은 어류에게 물리적 피해를 줄 수 있다. 이 연구는 최적의 수온(15℃)보다 높은 수온(20℃)에서의 대서양 연어의 건강상태를 평가하기 위해 수행하였다. 간 조직은 열 적응에 중요한 대사기능을 발휘하기에 본 연구에 간 조직을 사용하였다. 생체지표유전자의 개발을 위한 분석 방법으로는 NGS RNAseq 방법을 사용하였고, 생체지표유전자의 발현 양상을 관찰하기 위한 분석 방법으로는 RT-qPCR을 사용하였다. NGS RNAseq 분석을 통해 1,366개의 차별적 발현 유전자를 확인하였으며, 그 중에서 880개의 증가하는 유전자와 486개의 감소하는 유전자를 확인하였다. 생체지표유전자로는 heat shock protein 90 alpha (Hsp90α), heat shock protein 90 beta (Hsp90β) 및 cytochrome P450 1A (CYP1A)을 선정하였는데 이들 유전자는 NGS RNAseq 분석에서 수온의 변화에 민감하게 반응하는 유전자들이었다. 이들 유전자의 RT-qPCR을 통한 발현 양상은 NGS RNAseq 분석과 유사하게 나타났다. 이 연구의 결과는 다른 어종에도 적용할 수 있으며, 산업적으로도 유용하다고 생각된다.