• Title/Summary/Keyword: T-M interaction

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Curcumin and its Analogues (PGV-0 and PGV-1) Enhance Sensitivity of Resistant MCF-7 Cells to Doxorubicin through Inhibition of HER2 and NF-kB Activation

  • Meiyanto, Edy;Putri, Dyaningtyas Dewi Pamungkas;Susidarti, Ratna Asmah;Murwanti, Retno;Sardjiman, Sardjiman;Fitriasari, Aditya;Husnaa, Ulfatul;Purnomo, Hari;Kawaichi, Masashi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.179-184
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    • 2014
  • Chemoresistance of breast cancer to doxorubicin is mediated mainly through activation of NF-kB and over expression of HER2. Curcumin and its analogues (PGV-0 and PGV-1) exert cytotoxic effects on T47D breast cancer cells. Suppression of NF-kB activation is suggested to contribute to this activity. The present study aimed to explore the effects of curcumin, PGV-0, and PGV-1 singly and in combination with doxorubicin on MCF-7/Dox cells featuring over-expression of HER2. In MTT assays, curcumin, PGV-0, and PGV-1 showed cytotoxicity effects against MCF-7/Dox with IC50 values of $80{\mu}M$, $21{\mu}M$, and $82{\mu}M$ respectively. These compounds increased MCF-7/Dox sensitivity to doxorubicin. Cell cycle distribution analysis exhibited that the combination of curcumin and its analogues with Dox increased sub G-1 cell populations. Curcumin and PGV-1 but not PGV-0 decreased localization of p65 into the nucleus induced by Dox, indicating that activation of NF-kB was inhibited. Molecular docking of curcumin, PGV-0, and PGV-1 demonstrated high affinity to HER2 at ATP binding site. This interaction were directly comparable with those of ATP and lapatinib. These findings suggested that curcumin, PGV-0 and PGV-1 enhance the Dox cytotoxicity to MCF-7 cells through inhibition of HER2 activity and NF-kB activation.

PET Imaging of Click-engineered PSMA-targeting Immune Cells in Normal Mice

  • Hye Won Kim;Won Chang Lee;In Ho Song;Hyun Soo Park;Sang Eun Kim
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.8 no.2
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    • pp.53-61
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    • 2022
  • This study aimed to increase the targeting ability against PSMA in cell therapy using metabolic glycoengineering and biorthogonal chemistry and to visualize cell trafficking using PET imaging. Cellular membranes of THP-1 cells were decorated with azide(-N3) using Ac4ManNAz by metabolic glycoengineering. Engineered THP-1 cells were conjugated with DBCO-bearing fluorophore (ADIBO-Cy5.5) for 1 h at different concentrations and analyzed by confocal fluorescence microscopy and flow cytometry. For PSAM ligand conjugation to THP-1 cells, Ac4ManNAz treated THP-1 cells were incubated with DBCO-PSMA ligand (ADIBO-GUL) at a final concentration with 100 µM for 1 h. To evaluate the effect on cell recognition, PSMA ligand conjugated THP-1 cells(as effectors) were co-cultured with PSMA positive 22RV1 (as target cells) at 3 : 1 a effector-to-target cell (E/T) ratio. The interaction between THP-1 and 22RV1 was monitored by confocal fluorescence microscopy. For preparing the radiolabeled THP-1, the cells were treated at the activity of ~ 740 kBq of [89Zr]Zr(oxinate)4/5 × 106 cells. Radiolabeled cells were analyzed for determination of cell-associated radioactivity by gamma counting and viability using MTS assay. In the cytotoxicity assay, THP-1 cells did not have any cytotoxicity even when the Ac4ManNAz concentration was 100 µM. In confocal microscopy and flow cytometry, THP-1 cells were efficiently labeled ADIBO-Cy5.5 in a dose-dependent manner, and the dose of 100 µM was the optimal concentration for the following experiments. The clusters of PSMA ligand-conjugated THP-1 cells and 22RV1 cells were identified, indicating cell-cell recognition over the cell surface between two types of cells. Cell radiolabeling efficiency was 54.5 ± 17.8%. THP-1 labeled with 0.09 ± 0.03 Bq/cell showed no significant cytotoxicity compared to unlabeled THP-1 up to 7 days. We successfully demonstrated that Ac4ManNAz treated cells were efficiently conjugated with ADIBO-GUL for preparing the PSMA-targeting cells, and [89Zr]Zr(oxinate)4 could be used to label cells without toxicity. It suggested that PSMA-ligand conjugated cell therapy could be improved cell targeting and be monitored by PET imaging.

CDK2AP1, a Cyclin-Dependent Kinase 2-Associated Protein, Interacts with Kinesin-1 through Kinesin Superfamily Protein 5A (KIF5A) (Cyclin-dependent kinase 1 결합 단백질 CDK2AP1은 kinesin superfamily protein 5A (KIF5A)을 매개로 Kinesin-1와 결합)

  • Myoung Hun Kim;Se Young Pyo;Young Joo Jeong;Sung Woo Park;Mi Kyoung Seo;Won Hee Lee;Sang-Hwa Urm;Mooseong Kim;Jung Goo Lee;Dae-Hyun Seog
    • Journal of Life Science
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    • v.33 no.7
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    • pp.531-537
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    • 2023
  • Intracellular and axonal transport is mediated by microtubule-dependent motor proteins, such as kinesins and cytoplasmic dynein. Kinesin moves along the microtubule to the positive end of the microtubule, while dynein moves to the negative end of the microtubule. Kinesin-1 was first identified as a kinesin superfamily protein (KIF) that functions in the intracellular transport of various cargoes, including organelles, neurotransmitter receptors, and mRNA-protein complexes, through interactions between the carboxyl (C)-terminal domain and the cargo. It interacts with other cargoes, but the adapter/scaffold proteins that mediate between kinesin-1 and the cargo have yet to be fully identified. In this study, a yeast two-hybrid screen was used to identify adapter proteins that interact with the C-terminal region of KIF5A. We found an association between the C-terminal region of KIF5A and the cyclin-dependent kinase 2-associated protein 1 (CDK2AP1), originally identified in malignant hamster oral keratinocytes. CDK2AP1 bound to the C-terminal region of KIF5A and did not interact with KIF3A (the motor of kinesin-2), KIF5B, KIF5C, and kinesin light chain 1 (KLC1). The C-terminal region of CDK2AP1 is essential for its interaction with KIF5A. When co-expressed in HEK-293T cells, CDK2AP1 and kinesin-1 co-immunoprecipitated and co-localized in the cells. These results suggest that the KIF5A-CDK2AP1 interaction serves as an adapter protein connecting kinesin-1 and the cargo when kinesin-1 transports cargo in cells.

Removal of Red Tide Organisms -2. Flocculation of Red Tide Organisms by Using Loess- (적조생물의 구제 -2. 황토에 의한 적조생물의 응집제거-)

  • KIM Sung-Jae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.5
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    • pp.455-462
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    • 2000
  • The objective of this study was to examine the physicochemical characteristics of coagulation reaction between loess and red tide organisms (RTO) and its feasibility, in developing a technology for the removal of RTO bloom in coastal sea. The physicochemical characteristics of loess were examined for a particle size distribution, surface characteristics by scanning electron microscope, zeta potential, and alkalinity and pH variations in sea water. Two kinds of RTO that were used in this study, Cylindrothen closterium and Skeietonema costatum, were sampled in Masan bay and were cultured in laboratory. Coagulation experiments were conducted using various concentrations of loess, RTO, and a jar tester. The supernatant and RTO culture solution were analyzed for pH, alkalinity, RTO cell number. A negative zeta potential of loess increased with increasing pH at $10^(-3)M$ NaCl solution and had -71.3 mV at pH 9.36. Loess had a positive zeta potential of +1,8 mV at pH 1.98, which resulted in a characteristic of material having an amphoteric surface charge. In NaCl and $CaCl_2$, solutions, loess had a decreasing negative zeta potential with increasing $Na^+\;and\;Ca^(+2)$ ion concentration and then didn't result in a charge reversal due to not occurring specific adsorption for $Na^+$ ion while resulted in a charge reversal due to occurring specific adsorption for $Ca^(+2)$ ion. In sea water, loess and RTO showed the similar zeta potential values of -112,1 and -9.2 mV, respectively and sea sand powder showed the highest zeta potential value of -25.7 mV in the clays. EDLs (electrical double-layers) of loess and RTO were extremely compressed due to high concentration of salts included in sea water, As a result, there didn't almost exist EDL repulsive force between loess and RTO approaching each other and then LVDW (London-yan der Waals) attractive force was always larger than EDL repulsive force to easily form a floe. Removal rates of RTO exponentially increased with increasing a loess concentration. The removal rates steeply increased until $800 mg/l$ of loess, and reached $100{\%}$ at 6,400 mg/l of loess. Removal rates of RTO exponentially increased with increasing a G-value. This indicated that mixing (i.e., collision among particles) was very important for a coagulation reaction. Loess showed the highest RTO removal rates in the clays.

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SURFACE ROUGHNESS OF COMPOSITE RESIN ACCORDING TO FINISHING METHODS (복합레진 표면의 연마방법에 따른 표면조도)

  • Min, Jeong-Bum;Cho, Kong-Chul;Cho, Young-Gon
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.138-150
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    • 2007
  • The purpose of this study was to evaluate the difference of surface roughness of composite resin according to composite resin type, polishing methods, and use of resin sealant. Two hundred rectangular specimens, sized $8{\times}3{\times}2mm$, were made of Micro-new (Bisco, Inc., Schaumburg, IL, U.S.A) and Filtek Supreme (3M ESPE Dental Products, St. Paul, MN, U.S.A.), and divided into two groups; Micronew-M group, Filtek Supreme-S group. Specimens for each composite group were subdivided into five groups by finishing and polishing instruments used; M1 & S1(polyester strip), M2 & S2 (Sof-Lex disc), M3 & S3 (Enhance disc and polishing paste), M4 & S4(Astropol) and M5 & S5 (finishing bur), Polished groups were added letter B after the application of resin surface sealant (Biscover), eg, M1B and S1B. After specimens were stored with distilled water for 24hr, average surface roughness (Ra) was taken using a surface roughness tester. Representative specimens of each group were examined by FE-SEM (S-4700: Hitachi High Technologies Co., Tokyo, Japan). The data were analysed using paired t-test, ANOVA and Duncan's tests at the 0.05 probability level. The results of this study were as follows ; 1. The lowest Ra was achieved in all groups using polyester strip and the highest Ra was achieved in M5, S5 and M5B groups using finishing bur. On FE-SEM, M1 and S1 groups provided the smoothest surfaces, M5 and S5 groups were presented the roughest surfaces and voids by debonding of filler on the polished specimens. 2. There was no significant difference in Ra between Micronew and Filtek Supreme before the application of resin sealant, but Micronew was smoother than Filek Supreme after the application of resin sealant. 3. There was significant corelation between Ra of type of composite resin and polishing methods before the application of resin sealant (p=0.000), but no significant interaction between them after the application of resin sealant. On FE-SEM, most of composite resin surfaces were smooth after the application of resin sealant on the polished specimens. 4. Compared with before and after the application of resin sealant in group treated in the same composite and polishing methods, Ra of M4B and M5B was statistically lower than that of M4 and M5, and S5B was lower than that of S5, respectively (p<0.05). In conclusion, surface roughness by polishing instruments was different according to type of composite resin. Overall, polyester strip produced the smoothest surface, but finishing bur produced the roughest surface. Application of resin sealant provided the smooth surfaces in specimens polished with Enhance, Astropol and finishing bur, but not provided them in specimens polished with Sof-Lex disc.

A Matter of Autonomy in Art Criticism on Modernism (모더니즘 미술비평에 있어서 '자율성' (Autonomy)의 문제)

  • Choi Kwang-Jin
    • Journal of Science of Art and Design
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    • v.3
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    • pp.87-144
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    • 2001
  • This study looks into historical genealogy of autonomy in art criticism on modernism and presents the view of the judgment and correction about that. A matter of autonomy in art appeared in the attempt to totally reconsider and upset the theory of 'Mimesis' or 'Representation' which was the basis of traditional aesthetic theory. In the traditional theory of representation, they assumed primary image exists first and then tried to obtain visual similarity to it through art works. However, in the theory of autonomy in modernism, they maintained the reduction to pure form' or medium', regarding what art works represents and how similar to primary image are not the true essence of art. In the early 20th century, C. Bell laid the foundation stone of the theory of Formalism', providing that a matter of autonomy is significant form', which is the combination of lines and colors Aesthetic autonomy theory came to a climax by C. Greenberg, who systemized art criticism on modernism in the middle 20th century. According to his theory, the pursuit of the essence of form resulted in the specificity of medium' and flatness. They thought that the autonomy of art would be achieved by eliminating outward social factors from art works. This theory ended by Minimalism preventing the instructive function of art work and only emphasizing its material property. Since the middle 20th century, the autonomy theory was confronted with the limit and intense attack because it resulted in this fixed canon and materialism, so they began laying emphasis on those extrinsic factors around art works such as human life, society, history, and so on. This study focuses on arguing and complementing the limit of autonomy such as the adhesive and fixed canon, and then defining the more dynamic area of it. For this, first, I introduced the view of T. J. Clark and T. Crow who criticized the aesthetic autonomy theory. They denied the transcendental structure of form, and found form only in the association with substantial life and society. And they insisted the dynamism of form by emphasizing form as a result of negation insisted by avant-garde. Second, I researched the view of A. C, Danto and M. Fried, who complemented the traditional autonomy theory. They made autonomy emerge from the fixation of form like flatness through connecting essentialism with historical view. In conclusion, I insist that autonomic position of art make it possible to connect or mediate between material form and human or social elements. Therefore, autonomy should not be reduced to the axis of form or that of society but make interaction between two heterogeneous axes.

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Feline adipose tissue-derived mesenchymal stem cells pretreated with IFN-γ enhance immunomodulatory effects through the PGE2 pathway

  • Park, Seol-Gi;An, Ju-Hyun;Li, Qiang;Chae, Hyung-Kyu;Park, Su-Min;Lee, Jeong-Hwa;Ahn, Jin-Ok;Song, Woo-Jin;Youn, Hwa-Young
    • Journal of Veterinary Science
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    • v.22 no.2
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    • pp.16.1-16.13
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    • 2021
  • Background: Preconditioning with inflammatory stimuli is used to improve the secretion of anti-inflammatory agents in stem cells from variant species such as mouse, human, and dog. However, there are only few studies on feline stem cells. Objectives: This study aimed to evaluate the immune regulatory capacity of feline adipose tissue-derived (fAT) mesenchymal stem cells (MSCs) pretreated with interferon-gamma (IFN-γ). Methods: To assess the interaction of lymphocytes and macrophages with IFN-γ-pretreated fAT-MSCs, mouse splenocytes and RAW 264.7 cells were cultured with the conditioned media from IFN-γ-pretreated MSCs. Results: Pretreatment with IFN-γ increased the gene expression levels of cyclooxygenase-2, indoleamine 2,3-dioxygenase, hepatocyte growth factor, and transforming growth factor-beta 1 in the MSCs. The conditioned media from IFN-γ-pretreated MSCs increased the expression levels of M2 macrophage markers and regulatory T-cell markers compared to those in the conditioned media from naive MSCs. Further, prostaglandin E2 (PGE2) inhibitor NS-398 attenuated the immunoregulatory potential of MSCs, suggesting that the increased PGE2 levels induced by IFN-γ stimulation is a crucial factor in the immune regulatory capacity of MSCs pretreated with IFN-γ. Conclusions: IFN-γ pretreatment improves the immune regulatory profile of fAT-MSCs mainly via the secretion of PGE2, which induces macrophage polarization and increases regulatory T-cell numbers.

Effect of night light regimen on growth performance, antioxidant status and health of broiler chickens from 1 to 21 days of age

  • Zhao, R.X.;Cai, C.H.;Wang, P.;Zheng, L.;Wang, J.S.;Li, K.X.;Liu, W.;Guo, X.Y.;Zhan, X.A.;Wang, K.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.904-911
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    • 2019
  • Objective: The study was conducted to evaluate the effects of night light regimen on growth performance, antioxidant status and health of Lingnan Yellow broiler chickens from 1 to 21 days of age. Methods: A completely randomized factorial design involved 2 photoperiods (constant lighting [CL], 24 L:0 D and intermittent lighting [INL], 17 L:3 D:1 L:3 D)${\times}2$ light intensities (10 lx and 30 lx). A total of one thousand six hundred and eighty 1-d-old Lingnan Yellow broiler chicks were randomly divided into 4 treatments with 6 replicates (70 birds per replicate). The experiment lasted for 21 d. Results: Photoperiods and light intensities had no effect on average daily gain, feed conversion ratio, and mortality of the broiler chickens (p>0.05). The INL had a significant effect on average daily feed intake (p<0.05) of broiler chickens compared with CL. Photoperiod and light intensity had an interactive effect on melatonin (MT) concentration (p<0.05). At CL, reducing light intensity increased MT concentration; INL birds had higher MT but MT concentration was not affected by light intensity. There was an interactive effect on glutathione peroxidase (GPx) and catalase (CAT) in serum and total antioxidant capability (T-AOC) in liver between photoperiod and light intensity. With the decrease of light intensity, the activities of GPx and CAT in serum and T-AOC in liver increased in CL group (p<0.05). Broiler chickens reared under INL had better antioxidant status and 10 lx treatments had higher activities of CAT in serum than 30 lx (p<0.05). Different photoperiods and light intensities had no effect on malondialdehyde. There was an interaction between photoperiod and light intensity on serum creatine kinase (CK) concentration (p<0.05). At CL, the elevated light intensity resulted in an increase in CK content; INL birds had lower CK concentration especially in low light intensity group. Besides, INL and low light intensity significantly reduced the concentration of serum corticosterone and heat shock protein 70 (p<0.05). Serum immunoglobulin M contents were increased in broiler chickens reared under the INL compared with CL group (p<0.05). Conclusion: Results above suggest that the night light regimen of INL and 10 lx could be beneficial to the broiler chickens from 1 to 21 days of age due to the better health status and electricity savings.

Effect of Lidocaine-HCl on Microviscosity of Phosphatidylcholine Model Membrane

  • Chung, In-Kyo;Kim, Inn-Se;Choi, Chang-Hwa;Cho, Goon-Jae;Kim, Jin-Bom;Son, Woo-Sung;Jang, Hye-Ock;Yun, Il
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.3
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    • pp.243-251
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    • 2000
  • In order to provide a basis for studying the molecular mechanism of pharmacological action of local anesthetics and to develop a fluorescence spectroscopic method which can detect the microviscosity of native and model membranes using intramolecular excimerization of 1,3-di(l-pyrenyl)propane (Py-3-Py), we examined the effect of lidocaine HCl on the microviscosity of model membranes of phosphatidylcholine fraction extracted from synaptosomal plasma membrane vesicles (SPMVPC). The excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py in liquid paraffin was a simple linear function of $T/{\eta}.$ Based on this calibration curve, the microviscosity values of the direct probe environment in SPMVPC model membranes ranged from $234.97{\pm}48.85$ cP at $4^{\circ}C$ to %19.21{\pm}1.11$ cP at $45^{\circ}C.$ At $37^{\circ}C,$ a value of $27.25{\pm}0.44$ cP was obtained. The lidocaine HCl decreased the microviscosity of SPMVPC model membranes in a concentration-dependent manner, with a significant decrease in microviscosity value by injecting the local anesthetic even at the concentration of 0.5 mM. These results indicate that the direct environment of Py-3-Py in the SPMVPC model membranes is significantly fluidized by the lidocaine HCl. Also, the present study explicitly shows that an interaction between local anesthetics and membrane lipids is of importance in the molecular mechanism of pharmacological action of lidocaine HCl.

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Fermented ginseng extract, BST204, disturbs adipogenesis of mesenchymal stem cells through inhibition of S6 kinase 1 signaling

  • Yi, Sang Ah;Lee, Jieun;Park, Sun Kyu;Kim, Jeom Yong;Park, Jong Woo;Lee, Min Gyu;Nam, Ki Hong;Park, Jee Hun;Oh, Hwamok;Kim, Saetbyul;Han, Jihoon;Kim, Bo Kyung;Jo, Dong-Gyu;Han, Jeung-Whan
    • Journal of Ginseng Research
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    • v.44 no.1
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    • pp.58-66
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    • 2020
  • Background: The biological and pharmacological effects of BST204, a fermented ginseng extract, have been reported in various disease conditions. However, its molecular action in metabolic disease remains poorly understood. In this study, we identified the antiadipogenic activity of BST204 resulting from its inhibition of the S6 kinase 1 (S6K1) signaling pathway. Methods: The inhibitory effects of BST204 on S6K1 signaling were investigated by immunoblot, nuclear fractionation, immunoprecipitation analyses. The antiadipogenic effect of BST204 was evaluated by measuring mRNA levels of adipogenic genes and by chromatin immunoprecipitation and quantitative real-time polymerase chain reaction analysis. Results: Treatment with BST204 inhibited activation and nuclear translocation of S6K1, further decreasing the interaction between S6K1 and histone H2B in 10T1/2 mesenchymal stem cells. Subsequently, phosphorylation of H2B at serine 36 (H2BS36p) by S6K1 was reduced by BST204, inducing an increase in the mRNA expression of Wnt6, Wnt10a, and Wnt10b, which disturbed adipogenic differentiation and promoted myogenic and early osteogenic gene expression. Consistently, BST204 treatment during adipogenic commitment suppressed the expression of adipogenic marker genes and lipid drop formation. Conclusion: Our results indicate that BST204 blocks adipogenesis of mesenchymal stem cells through the inhibition of S6K1-mediated histone phosphorylation. This study suggests the potential therapeutic strategy using BST204 to combat obesity and musculoskeletal diseases.