• BMB Reports
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• v.45 no.6
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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.

• Proceedings of the Zoological Society Korea Conference
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• 2001.10a
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• pp.212.2-213
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• 2001

No Abstract, See Full Text

• Journal of Pharmacopuncture
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• v.22 no.1
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• pp.28-34
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• 2019

Background: Breast cancer is a complex, heterogeneous disease and one of the most common malignancies in women worldwide. The efficacy of chemotherapy as an important breast cancer treatment option has been severely limited because of the inherent or acquired resistance of cancer cells. The molecular chaperone heat shock protein 90 (HSP90) upregulated in response to cellular stress is required for functions such as conformational maturation, activation and stability in more than 200 client proteins, mostly of the signaling type. In this study, the expression of HSP90 isoforms including $HSP90{\alpha}$ and $HSP90{\beta}$ in breast cancer cell lines before and after treatment with doxorubicin (DOX) was assessed. Material and Methods: The cell cytotoxicity of DOX in MDA-MB-231 and MCF-7 cell lines was determined using the MTT assay. immunofluorescence and western blotting techniques were used to determine the expression of $HSP90{\beta}$ in the cell lines before and after DOX treatment. Immunofluorescence was also conducted to ascertain the expression of $HSP90{\alpha}$. Results: The MTT assay results showed that the MDA-MB-231 cells ($IC_{50}=14.521{\mu}M$) were more sensitive than the MCF-7 cells ($IC_{50}=16.3315{\mu}M$) to DOX. The immunofluorescence results indicated that the expression of $HSP90{\alpha}$ in both cell lines decreased after exposure to DOX. The western blot and immunofluorescence analyses showed that $HSP90{\beta}$ expression decreased in the MCF-7 cells but increased in the MDA-MB-231 cells after DOX treatment. Conclusion: The obtained results suggested that $HSP90{\alpha}$ and $HSP90{\beta}$ expression levels were reduced in the MCF-7 cells after exposure to DOX. In the MDA-MB-231 cells, $HSP90{\alpha}$ expression was reduced while $HSP90{\beta}$ was found to be overexpressed following DOX treatment.

• Journal of Life Science
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• v.18 no.12
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• pp.1637-1643
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• 2008

This study has investigated whether extracellular HSP90 predisposes vascular smooth muscle cells (VSMCs) to pro-inflammatory phenotype. Exposure of rat aortic smooth muscle cells to HSP90 not only enhanced IL-6 release but also profoundly induced IL-6 transcript via promoter activation. HSP90-induced IL-6 promoter activation was suppressed by dominant-negative forms of Toll-like receptor (TLR)-4 and myeloid differentiation factor 88 (MyD88), but not by dominant-negative-forms of TLR-3 and TIR-domain-containing adapter-inducing interferon-${\beta}$ (TRIF). Curcumin, which inhibits dimerization of TLR-4, also attenuated the IL-6 induction by HSP90. Mutation at the NF-${\kappa}B$- or C/EBP-binding site in the IL-6 promoter region suppressed the promoter activation in response to HSP90. The gene delivery of $I{\kappa}B$ using recombinant adenoviruses and treatment with resveratrol, which inhibit NF-${\kappa}B$ activity, attenuated the HSP90-induced IL-6 release from VSMCs. The present study proposes that extracellular HSP90 would contribute to inflammatory reaction in the stressed vasculature by inducing IL-6 in VSMCs, and that TLR-4 and NF-${\kappa}B$ would play active roles in the process.

• Journal of Animal Science and Technology
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• v.64 no.6
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• pp.1046-1062
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• 2022

This study investigated the effects of L-glutamine (Gln) supplementation on growth performance, physiological traits, heat shock proteins (HSPs), and gene expression related to muscle and adipose tissue development in Hanwoo steers under heat stress (HS) conditions. Eight Hanwoo steers (initial body weight [BW] 570.7 ± 43.6 kg, months of age 22.3 ± 0.88) were randomly separated into two groups, control and treatment, and supplied with the concentration (1.5% of BW kg/day/head) and rice straw (1.5 kg/day/head). The treatment group were fed the Gln supplementation (0.5% of concentration, as-fed basis) once a day at 08:00 h. Blood samples for the assessment of haematological and biochemical parameters and the separation of peripheral blood mononuclear cells (PBMCs) were collected four times, at 0, 3, 6, and 10 weeks of the experiment. Feed intake was measured daily. BW to analyze growth performance and hair follicle collection to analyze the expression of HSPs were executed four times at 0, 3, 6, and 10 weeks. To analyze gene expression, longissimus dorsi muscle samples were collected by biopsy at the end of the study. As a result, growing performance, including final BW, average daily gain, and gain-to-feed ratio, were not different between the two groups. Leukocytes including lymphocytes and granulocytes, tended to increase in the Gln supplementation group (p = 0.058). There were also no differences in biochemical parameters shown between the two groups, except total protein and albumin, both of which were lower in the Gln supplementation group (p < 0.05). Gene expressions related to muscle and adipose tissue development were not different between the two groups. As temperature-humidity index (THI) increased, HSP70 and HSP90 expression in the hair follicle showed a high correlation. HSP90 in the hair follicle was decreased in the treatment group compared with the control group at 10 weeks (p < 0.05). Collectively, dietary Gln supplementation (0.5% of concentration, as-fed basis) may not be influential enough to affect growth performance and gene expression related to muscle and adipose tissue development in steers. However, Gln supplementation increased the number of immune cells and decreased HSP90 in the hair follicle implying HS reduction in the corresponding group.

• Biomolecules & Therapeutics
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• v.27 no.5
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• pp.423-434
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• 2019

HSP90 is a molecular chaperone that increases the stability of client proteins. Cancer cells show higher HSP90 expression than normal cells because many client proteins play an important role in the growth and survival of cancer cells. HSP90 inhibitors mainly bind to the ATP binding site of HSP90 and inhibit HSP90 activity, and these inhibitors can be distinguished as ansamycin and non-ansamycin depending on the structure. In addition, the histone deacetylase inhibitors inhibit the activity of HSP90 through acetylation of HSP90. These HSP90 inhibitors have undergone or are undergoing clinical trials for the treatment of cancer. On the other hand, recent studies have reported that various reagents induce cleavage of HSP90, resulting in reduced HSP90 client proteins and growth suppression in cancer cells. Cleavage of HSP90 can be divided into enzymatic cleavage and non-enzymatic cleavage. Therefore, reagents inducing cleavage of HSP90 can be classified as another class of HSP90 inhibitors. We discuss that the cleavage of HSP90 can be another mechanism in the cancer treatment by HSP90 inhibition.

• Journal of Acupuncture Research
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• v.24 no.6
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• pp.29-36
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• 2007

Electroacupuncture(EA) is known to affect various autonomic functions such as blood pressure regulation, immune modulation and the improvement of disorders concerning autonomic functions. The aim of the present study was to establish whether EA has an immune-enhancing effect. ICR mice weighing 20 to 25g were divided into four groups: Group I(n=6), blank; Group II(n=6), control; Group III(Zusanli, n=6), low frequency-EA(10Hz of electrical stimulation), and Group IV(Zusanli, n=6), high frequency-EA (100Hz of electrical stimulation). For this study, we investigated expressions of spleen heat shock protein (HSP)70, HSP90 and secretions of cytokines. A Forced Swimming Test(FST) was performed as a model of activity test in mice. After three days of the FST, 10Hz EA($114.8{\pm}7.27s$) and 100Hz EA($147.5{\pm}1.29s$) immobility time significantly decreased compared with the control group($157.2{\pm}1.48s$). After seven days, 10Hz EA($124{\pm}1s$) and 100Hz EA($141{\pm}4.24s$) also significantly decreased immobility time compared with the control group($168{\pm}7.93s$). 10Hz EA and 100Hz EA increased the expression of HSP70 but did not change that of HSP90. 100Hz EA increased secretions of IL-6 and IL-12 compared with the control group however, 10Hz EA failed to change those of IL-6 and IL-12. The present results suggest that EA may be useful for down-regulated immune diseases.

• 대한생식의학회:학술대회논문집
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• 2006.06a
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• pp.150.2-150.2
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• 2006

• Journal of Animal Science and Technology
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• v.64 no.1
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• pp.110-122
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• 2022

The reduction of milk yield caused by heat stress in summer is the main condition restricting the economic benefits of dairy farms. To examine the impact of hyperthermia on bovine mammary epithelial (MAC-T) cells, we incubated the MAC-T cells at thermal-neutral (37℃, CON group) and hyperthermic (42℃, HS group) temperatures for 6 h. Subsequently, the cell viability and apoptotic rate of MAC-T cells, apoptosis-related genes expression, casein and amino acid transporter genes, and the expression of the apoptosis-related proteins were examined. Compared with the CON group, hyperthermia significantly decreased the cell viability (p < 0.05) and elevated the apoptotic rate (p < 0.05) of MAC-T cells. Moreover, the expression of heat shock protein (HSP)70, HSP90B1, Bcl-2-associated X protein (BAX), Caspase-9, and Caspase-3 genes was upregulated (p < 0.05). The expression of HSP70 and BAX (pro-apoptotic) proteins was upregulated (p < 0.05) while that of B-cell lymphoma (BCL)2 (antiapoptotic) protein was downregulated (p < 0.05) by hyperthermia. Decreased mRNA expression of mechanistic target of rapamycin (mTOR) signaling pathway-related genes, amino acid transporter genes (SLC7A5, SLC38A3, SLC38A2, and SLC38A9), and casein genes (CSNS1, CSN2, and CSN3) was found in the heat stress (HS) group (p < 0.05) in contrast with the CON group. These findings illustrated that hyperthermia promoted cell apoptosis and reduced the transport of amino acids into cells, which inhibited the milk proteins synthesis in MAC-T cells.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.11
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• pp.4475-4482
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• 2014

Background: Cisplatin, a DNA damaging agent, induces apoptosis through increasing DNA fragmentation. However, identification of intrinsic resistance molecules against Cisplatin is vital to estimate the success of therapy. Bag-1 (Bcl-2-associated anthanogene) is one anti-apoptotic protein involved in drug resistance impacting on therapeutic efficiency. Elevated levels of this protein are related with increase cell proliferation rates, motility and also cancer development. For this reason, we aimed to understand the role of Bag-1 expression in Cisplatin-induced apoptosis in HeLa cervix cancer cells. Cisplatin decreased cell viability in time- and dose-dependent manner in wt and Bag-1L+HeLa cells. Although, $10{\mu}M$ Cisplatin treatment induced cell death within 24h by activating caspases in wt cells, Bag-1L stable transfection protected cells against Cisplatin treatment. To assess the potential protective role of Bag-1, we first checked the expression profile of interacting anti-apoptotic partners of Bag-1. We found that forced Bag-1L expression prevented Cisplatin-induced apoptosis through acting on Mcl-1 expression, which was reduced after Cisplatin treatment in wt HeLa cells. This mechanism was also supported by the regulation of heat shock protein (Hsp) family members, Hsp90 and Hsp40, which were involved in the regulation Bag-1 interactome including several anti-apoptotic Bcl-2 family members and c-Raf.