동결보존이 생쥐 난소 조직 내 Heat Shock Protein 90의 발현에 미치는 영향

이선희,박용석,염혜원,송견지,한상철,배인하,Lee, Sun-Hee,Park, Yong-Seog,Yeum, Hye-Won,Song, Gyun-Jee,Han, Sang-Chul,Bae, In-Ha 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.1

Objective : Heat shock protein family is related to protective mechanism of cells by environmental changes. This study was performed to evaluate the effect of cryopreservation on the heat shock protein 90 (Hsp90) expression in mouse ovarian tissue. Methods : Cryopreservation of mouse ovarian tissue was carried out by slow freezing method. The mRNA level of Hsp90 expression in both fresh and cryopreserved mouse ovarian tissue was analyzed by RT-PCR. The protein expression of Hsp90 was evaluated by Western blot analysis and immunohistochemistry. Results: The mRNA and protein of Hsp90 were expressed in both fresh and cryopreserved mouse ovarian tissue. The amount of Hsp90 mRNA was increased in cryopreserved ovarian tissue after 60 and 90 minutes after thawing and incubation. The amount of Hsp90 protein was increased in the cryopreserved ovarian tissue after 6 hours of the incubation in Western blot analysis. In immunohistochemical study, Hsp90 protein was localized in cytoplasm of oocytes and granulosa cells. Significant level of immunoreactive Hsp90 protein was detected in theca cells contrast to the weak expression in ovarian epithelial cells. Conclusion: This results showed the increase of Hsp90 expression in both mRNA and protein level in the cryopreserved mouse ovarian tissue. It can be suggested that Hsp90 may play a role in the protective or recovery mechanism against the cell damage during cryopreservaion.

Small Molecule Inhibitors toDisrupt Protein-protein Interactions of Heat Shock Protein 90 Chaperone Machinery

Young Ho Seo 대한암예방학회 2015 Journal of cancer prevention Vol.20 No.1

Heat shock protein 90 (Hsp90) is an adenosine triphosphate dependent molecular chaperone in eukaryotic cells that regulates theactivation and maintenance of numerous regulatory and signaling proteins including epidermal growth factor receptor, human epidermal growth factor receptor 2, mesenchymal-epithelial transition factor, cyclin-dependent kinase-4, protein kinase B, hypoxia-inducible factor 1α, and matrix metalloproteinase-2. Since many of Hsp90 clients are oncogenic proteins, Hsp90 has become an attractive therapeutic target for treatment of cancer. To discover small molecule inhibitors targeting Hsp90 chaperone machinery, several strategies have been employed, which results in three classes of inhibitors such as N-terminal inhibitors, C-terminal inhibitors, and inhibitors disrupting protein-protein interactions of Hsp90 chaperone machinery. Developing small molecule inhibitors that modulate protein-protein interactions of Hsp90 is a challenging task, although it offers many alternative opportunities for therapeutic intervention. The lack of well-definedbinding pocket and starting points for drug design challenges medicinal chemists to discover small molecule inhibitors disrupting protein-protein interactions of Hsp90. The present review will focus on the current studies on small molecule inhibitors disrupting protein-protein interactions of Hsp90 chaperone machinery, provide biological background on the structure, function and mechanism of Hsp90’s protein-protein interactions, and discuss the challenges and promise of its small molecule modulations.

Apoptosis in response to heat stress is positively associated with heat-shock protein 90 expression in chicken myocardial cells in vitro

Xiao-Hui Zhang,Hong Wu,Shu Tang,Qiao-Ning Li,Jiao Xu,Miao Zhang,Ya-Nan Su,Bin Yin,Qi-Ling Zhao,Nicole Kemper,Joerg Hartung,Endong Bao 대한수의학회 2017 Journal of Veterinary Science Vol.18 No.2

To determine heat-shock protein (Hsp)90 expression is connected with cellular apoptotic response to heat stress and its mechanism, chicken (Gallus gallus) primary myocardial cells were treated with the Hsp90 promoter, aspirin, and its inhibitor, geldanamycin (GA), before heat stress. Cellular viability, heat-stressed apoptosis and reactive oxygen species level under different treatments were measured, and the expression of key proteins of the signaling pathway related to Hsp90 and their colocalization with Hsp90 were detected. The results showed that aspirin treatment increased the expression of protein kinase B (Akt), the signal transducer and activator of transcription (STAT)-3 and p-IKKa/b and the colocalization of Akt and STAT-3 with Hsp90 during heat stress, which was accompanied by improved viability and low apoptosis. GA significantly inhibited Akt expression and p-IKKa/b level, but not STAT-3 quantity, while the colocalization of Akt and STAT-3 with Hsp90 was weakened, followed by lower cell viability and higher apoptosis. Aspirin after GA treatment partially improved the stress response and apoptosis rate of tested cells caused by the recovery of Akt expression and colocalization, rather than the level of STAT-3 (including its co-localization with Hsp90) and p-IKKa/b. Therefore, Hsp90 expression has a positive effect on cellular capacity to resist heat-stressed injury and apoptosis. Moreover, inhibition of Hsp90 before stress partially attenuated its positive effects.

Analysis of the Correlation between Expressions of HSP90α, HSP90β, and GRP94, and the Clinicopathologic Characteristics in Tissues of Non-Small Cell Lung Cancer Patients

김미경,Kim, Mi Kyeong Korean Society for Clinical Laboratory Science 2017 대한임상검사과학회지(KJCLS) Vol.49 No.4

Heat shock proteins (HSPs) are induced as a self-defense mechanism of cells when exposed to various external stresses, such as high fever, infection, free radicals, and heavy metals. They affect the prognosis in the process of tumor formation. HSP is classified into four families: HSP27, HSP60, HSP90, and HSP100, depending on molecular weight. Heat shock protein 90 (HSP90), a molecular chaperone, plays an important role in the cellular protection against various stressful stimuli and in the regulation of cell cycle progression and apoptosis. In the present study, we assessed the differential expression of HSP90 family proteins in non-small cell lung cancer (NSCLC), and the correlation of their expression levels with clinicopathologic factors and patient survival rates. The result of this study can be summarized as follows; $HSP90{\alpha}$ showed higher expression in patients with no lymphovascular invasion (p=0.014). $HSP90{\beta}$ showed a higher expression of squamous cell carcinoma (p=0.003), and an over expression of glucose-related protein (GRP94) was significantly associated with poor differentiation (p=0.048). However, none of the HSP90 proteins showed a significant association with the survival status in patients with NSCLC. This study also indicates that $HSP90{\alpha}$ might contribute more to the carcinogenesis of NSCLC than $HSP90{\beta}$, and GRP94 and isoform selectivity should be considered when HSP90 inhibitors are studied or utilized in the treatment of NSCLC.

Analysis of the Correlation between Expressions of HSP90α, HSP90β, and GRP94, and the Clinicopathologic Characteristics in Tissues of Non-Small Cell Lung Cancer Patients

김미경 대한임상검사과학회 2017 대한임상검사과학회지(KJCLS) Vol.49 No.4

Heat shock proteins (HSPs) are induced as a self-defense mechanism of cells when exposed to various external stresses, such as high fever, infection, free radicals, and heavy metals. They affect the prognosis in the process of tumor formation. HSP is classified into four families: HSP27, HSP60, HSP90, and HSP100, depending on molecular weight. Heat shock protein 90 (HSP90), a molecular chaperone, plays an important role in the cellular protection against various stressful stimuli and in the regulation of cell cycle progression and apoptosis. In the present study, we assessed the differential expression of HSP90 family proteins in non-small cell lung cancer (NSCLC), and the correlation of their expression levels with clinicopathologic factors and patient survival rates. The result of this study can be summarized as follows; HSP90α showed higher expression in patients with no lymphovascular invasion (p=0.014). HSP90β showed a higher expression of squamous cell carcinoma (p=0.003), and an over expression of glucose-related protein (GRP94) was significantly associated with poor differentiation (p=0.048). However, none of the HSP90 proteins showed a significant association with the survival status in patients with NSCLC. This study also indicates that HSP90α might contribute more to the carcinogenesis of NSCLC than HSP90β, and GRP94 and isoform selectivity should be considered when HSP90 inhibitors are studied or utilized in the treatment of NSCLC.

Structural and functional differences of cytosolic 90-kDa heat-shock proteins (Hsp90s) in Arabidopsis thaliana

Cha, J.Y.,Ahn, G.,Kim, J.Y.,Kang, S.B.,Kim, M.R.,Su'udi, M.,Kim, W.Y.,Son, D. Gauthier-Villars ; Elsevier Science Ltd 2013 Plant physiology and biochemistry Vol.70 No.-

The seven members of the 90-kDa heat shock protein (Hsp90) family encode highly conserved molecular chaperones essential for cell survival in Arabidopsis thaliana. Hsp90 are abundant proteins, localized in different compartments with AtHsp90.1-4 in the cytosol and AtHsp90.5-7 in different organelles. Among the AtHsp90, AtHsp90.1, is stress-inducible and shares comparatively low sequence identity with the constitutively expressed AtHsp90.2-4. Even though abundant information is available on mammalian cytosolic Hsp90 proteins, it is unknown whether cytosolic Hsp90 proteins display different structural and functional properties. We have now analyzed two A. thalianas cytosolic Hsp90s, AtHsp90.1 and AtHsp90.3, for functional divergence. AtHsp90.3 showed higher holdase chaperone activity than AtHsp90.1, although both AtHsp90s exhibited effective chaperone activity. Size-exclusion chromatography revealed different oligomeric states distinguishing the two Hsp90 proteins. While AtHsp90.1 exists in several oligomeric states, including monomers, dimers and higher oligomers, AtHsp90.3 exists predominantly in a high oligomeric state. High oligomeric state of AtHsp90.1 showed higher holdase chaperone activity than the respective monomer or dimer states. When high oligomeric forms of AtHsp90.1 and AtHsp90.3 are reduced by DTT, activity was reduced compared to that found in the native high oligomeric state. In addition, ATP-dependent foldase chaperone activity of AtHsp90.3 was higher with strong intrinsic ATPase activity than that of AtHsp90.1. As a conclusion, the two A. thaliana cytosolic Hsp90 proteins display different functional activities depending on structural differences, implying functional divergence although the proteins are localized to the same sub-cellular organelle.

갑상선 종양에서 Heat Shock Protein70과 Heat Shock Protein90의 발현 양상

최진욱,김진영,박철영<SUP>1<.SUP>,오기원<SUP>1<.SUP>,임성희<SUP>1<.SUP>,박성우<SUP>1<.SUP>,조현득<SUP>2<.SUP>,이명준<SUP>3<.SUP>,김이수,Jin Wook Choi,M.D.,Jin Yong Kim,Cheol Young Park,M.D.<SUP>1<.SUP>,Ki Won Oh,M.D.<SUP>1<.SUP> 대한갑상선-내분비외과학회 2004 The Koreran journal of Endocrine Surgery Vol.4 No.2

Purpose: Heat shock proteins (hsps) are synthesized by cells in response to various stress conditions, including carcinogenesis. The expression of hsps in neoplasia has been implicated in the regulation of cell signaling pathway such as cell survival and apoptosis. This study aimed to determine whether hsps expression in various thyroid neoplasia are significant and to identify the possibility as a therapeutic molecular target. Methods: We examined the expression of the hsp70 and hsp90 on tissue section from 53 thyroid tissues (16 normal tissues; 11 nodular hyperplasia; 12 follicular adenomas; 14 papillary carcinomas) using immunohistochemistry. Hsps expression was scored according to the percentage of positively stained cells (grade 0 to grade III). Results: For hsp70, all of the 53 tissues showed over- expression. 100% (16/16) of normal thyroid tissue and 87.0% (20/23) of benign tissue were categorized as grade I or II. In comparison, the carcinoma tissues showed expression in 64.3% with grade III. For hsp90, almost of normal thyroid tissue and benign tumors showed no expression (87.5% in normal tissues, 91.3% in benign tumors). However, all of carcinoma tissues showed expression and 78.6% (11/14) of carcinoma were in grade II or III. Conclusion: In current study, the pattern of expression for hsp70 and hsp90 in normal, benign, malignant thyroid tissues suggests that heat shock proteins might have some role in tumorigenesis in thyroid. Since there have been no reports on heat shock proteins and thyroid, further study is necessary and could give us clinically significant clue for diagnosis and treatment. (Korean J Endocrine Surg 2004; 4:79-84)

Contribution of HSP90 Cleavage to the Cytotoxic Effect of Suberoylanilide Hydroxamic Acid In Vivo and the Involvement of TXNIP in HSP90 Cleavage

( Sangkyu Park ),( Dongbum Kim ),( Haiyoung Jung ),( In Pyo Choi ),( Hyung-joo Kwon ),( Younghee Lee ) 한국응용약물학회 2024 Biomolecules & Therapeutics(구 응용약물학회지) Vol.32 No.1

Heat shock protein (HSP) 90 is expressed in most living organisms, and several client proteins of HSP90 are necessary for cancer cell survival and growth. Previously, we found that HSP90 was cleaved by histone deacetylase (HDAC) inhibitors and proteasome inhibitors, and the cleavage of HSP90 contributes to their cytotoxicity in K562 leukemia cells. In this study, we first established mouse xenograft models with K562 cells expressing the wild-type or cleavage-resistant mutant HSP90β and found that the suppression of tumor growth by the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) was interrupted by the mutation inhibiting the HSP90 cleavage in vivo. Next, we investigated the possible function of thioredoxin interacting protein (TXNIP) in the HSP90 cleavage induced by SAHA. TXNIP is a negative regulator for thioredoxin, an antioxidant protein. SAHA transcriptionally induced the expression of TXNIP in K562 cells. HSP90 cleavage was induced by SAHA also in the thymocytes of normal mice and suppressed by an anti-oxidant and pan-caspase inhibitor. When the thymocytes from the TXNIP knockout mice and their wild-type littermate control mice were treated with SAHA, the HSP90 cleavage was detected in the thymocytes of the littermate controls but suppressed in those of the TXNIP knockout mice suggesting the requirement of TXNIP for HSP90 cleavage. We additionally found that HSP90 cleavage was induced by actinomycin D, β-mercaptoethanol, and p38 MAPK inhibitor PD169316 suggesting its prevalence. Taken together, we suggest that HSP90 cleavage occurs also in vivo and contributes to the anti-cancer activity of various drugs in a TXNIP-dependent manner.

Discovery and development of heat shock protein 90 inhibitors as anticancer agents: a review of patented potent geldanamycin derivatives

Kim, TaeHun,Keum, Gyochang,Pae, Ae Nim Informa UK, Ltd. 2013 Expert opinion on therapeutic patents Vol.23 No.8

<P><B><I>Introduction:</I></B> There has been research on anticancer strategies which focus on disrupting a single malignant protein. One of the strategies is the inhibition of one protein, heat shock protein 90 (Hsp90). There are many reasons why Hsp90 protein is targeted by anticancer agents: maintenance of cellular homeostasis in organisms involves Hsp90 and its client proteins; moreover, Hsp90 complex is involved in regulating several signal transduction pathways and plays an important role in the maturation of lots of tumor-promoting client proteins. Geldanamycin (GM), the first benzoquinone ansamycin, has shown anticancer activity by binding to Hsp90. Currently, several GM derivatives such as 17-AAG, 17-(2-dimethylaminoethyl)amino-17-demethoxygeldanamycin, IPI-493, and IPI-504 are being progressively developed toward clinical application.</P><P><B><I>Areas covered:</I></B> Several research groups have studied GM and its derivatives to develop novel and potent Hsp90 inhibitors for the treatment of cancer. The crystal structure of Hsp90 was utilized to undergo structural optimization of GM derivatives. A wide variety of structural modifications were performed and some of the derivatives are now in clinical studies. The aim of this review was to summarize and analyze the structure-activity relationships of GM derivatives and the focus is on patented novel and pharmaceutically efficacious derivatives published from 1971 to 2012.</P><P><B><I>Expert opinion:</I></B> Hsp90 inhibitors offer an effective therapeutic approach for treatment of cancer. To date, the clinical results of 17-AAG, IPI-493, and IPI-504 suggest that these GM derivatives could be used either alone or in combination with other marketed medications for the treatment of cancer patients. As there are not any marketed Hsp90 inhibitors, inhibiting Hsp90 chaperone function remains as a promising strategy that still requires further research.</P>

Heat shock protein 90 facilitates formation of the HBV capsid via interacting with the HBV core protein dimers

Shim, H.Y.,Quan, X.,Yi, Y.S.,Jung, G. Academic Press 2011 Virology Vol.410 No.1

The mechanism by which host factors contribute to hepatitis B virus (HBV) capsid formation during the viral life cycle remains unclear. This study analyzed the interaction between heat shock protein 90 (Hsp90), a host factor, and the HBV core protein. Hsp90 was found to bind to HBV core protein dimers, which was then encapsidated into the HBV capsid. Furthermore, activated Hsp90 may facilitate the formation of the human HBV capsid by catalyzing core assembly and reducing the degree of capsid dissociation at various temperatures, both in vitro and in vivo, and when subjected to detergent treatments in vitro. In addition, inhibition or downregulation of Hsp90 reduced HBV production in HepG2.2.15 cells. These results showed that Hsp90 plays an important role in HBV capsid stabilization and HBV formation.