Benzoxathiol derivative BOT-4-one suppresses L540 lymphoma cell survival and proliferation via inhibition of JAK3/STAT3 signaling

김병학,예상규,Yun Sook Min,Jung Sook Choi,Gyeong-Hun Baeg,Youngsoo Kim,신종욱,김태윤 생화학분자생물학회 2011 Experimental and molecular medicine Vol.43 No.5

Persistently activated JAK/STAT3 signaling pathway plays a pivotal role in various human cancers including major carcinomas and hematologic tumors, and is implicated in cancer cell survival and proliferation. Therefore, inhibition of JAK/STAT3 signaling may be a clinical application in cancer therapy. Here, we report that 2-cyclohexylimino-6-methyl-6,7-dihydro-5H-benzo [1,3]oxathiol-4-one (BOT-4-one), a small molecule inhibitor of JAK/STAT3 signaling, induces apoptosis through inhibition of STAT3 activation. BOT-4-one suppressed cytokine (upd)-induced tyrosine phosphorylation and transcriptional activity of STAT92E,the sole Drosophila STAT homolog. Consequently,BOT-4-one significantly inhibited STAT3 tyrosine phosphorylation and expression of STAT3 downstream target gene SOCS3 in various human cancer cell lines, and its effect was more potent in JAK3-activated Hodgkin’s lymphoma cell line than in JAK2-activated breast cancer and prostate cancer cell lines. In addition, BOT-4-one-treated Hodgkin’s lymphoma cells showed decreased cell survival and proliferation by inducing apoptosis through down-regulation of STAT3 downstream target anti-apoptotic gene expression. These results suggest that BOT-4-one is a novel small molecule inhibitor of JAK3/STAT3 signaling and may have therapeutic potential in the treatment of human cancers harboring aberrant JAK3/STAT3 signaling,specifically Hodgkin’s lymphoma.

Benzo[a]pyrene-induced Modification on p53 and Related Proteins

이순미,예상규,최진희,Lee Sun-Mi,Ye Sang-Kyu,Choi Jinhee The Korean Society of Environmental Toxicology 2005 환경독성보건학회지 Vol.20 No.1

PAH 위해성 평가의 생체지표 개발을 위하여, benzo[a]pyrene을 인체 간암 세포주인 HepG2세포에 처리하여 암 억제 단백질인 p53 및 관련 단백질의 발현 양상에 대하여 연구하였다. HepG2 세포의 생존력은 benzo[a]pyrene을 노출시킨 군에서 농도가 증가할수록 감소하였다. p53과 인산화 p53의 발현 양상은 benzo[a]pyrene 농도 의존적으로 증가하는 경향을 보였으며, 반면에 아세틸화 p53은 benzo[a]pyrene의 농도가 증가할수록 감소하는 경향을 나타내었다. 세포 주기 조절에 관련된 p21 단백질은 화학 물질 처리에 의해서 p53과 마찬가지로 증가하였으나, CdK4와 Rb 단백질의 발현에는 변화가 없었다. 상관분석 결과 Benzo[a]pyrene 노출, 세포 생존력, p53, 인산화 p53, p21이 서로 높은 상관성을 보였다. 본 연구의 결과는 p53 단백질의 축적이 benzo[a]pyrene 독성에 있어 매우 중요한 현상이며, 이는 선택적인 지표와 함께 p53 이 benzo[a]pyrene과 같은 PAH 계열의 물질의 위해성 평가를 위한 민감한 생체 지표로써 개발될 수 있음을 시사한다.

Chronic Non-Social Stress Affects Depressive Behaviors But Not Anxiety in Mice

윤상호,김병학,예상규,김명환 대한약리학회 2014 The Korean Journal of Physiology & Pharmacology Vol.18 No.3

The etiology of most psychiatric disorders is still incompletely understood. However, growing evidencesuggests that stress is a potent environmental risk factor for depression and anxiety. In rodents,various stress paradigms have been developed, but psychosocial stress paradigms have received moreattention than non-social stress paradigms because psychosocial stress is more prevalent in humans. Interestingly, some recent studies suggest that chronic psychosocial stress and social isolation affectsmainly anxiety-related behaviors in mice. However, it is unclear whether chronic non-social stressinduces both depression- and anxiety-related phenotypes or induces one specific phenotype in mice. In the present study, we examined the behavioral consequences of three chronic non-social stressparadigms: chronic predictable (restraint) stress (CPS), chronic unpredictable stress (CUS), andrepeated corticosterone-HBC complex injection (RCI). Each of the three paradigms induced mild tosevere depression/despair-like behaviors in mice and resulted in increased immobility in a tailsuspension test. However, anxiety-related phenotypes, thigmotaxis and explorative behaviors, were notchanged by the three paradigms. These results suggest that depression- and anxiety-related phenotypescan be dissociated in mouse stress models and that social and non-social stressors might affect braincircuits and behaviors differently.

벤조피렌에 의한 p53 및 관련 단백질 변화

최진희,이순미,예상규 환경독성보건학회 2005 환경독성보건학회지 Vol.20 No.1

PAH 위해성 평가의 생체지표 개발을 위하여, benzo[a]pyrene을 인체 간암 세포주인 HepG2 세포에 처리 하여 암 억제 단백질인 p53 . HepG2 세포의 생존력은 benzo[a]pyrene을 노출 시킨 군에서 농도가 증가 할수록 감소하였다. p53과 인산화 p53의 발현 양상은benzo[a]pyrene 농도 의존적으로 증가하는 경향을 보였으며, 반면에 아세틸화 p53은 benzo[a]pyrene의 농. 세포 주기 조절에 관련된 p21 단백질은 화학 물질 처리에의해서 p53과 마찬 가지로 증가하였으나, CdK4와 Rb 단백질의 발현에는 변화가 없었다. 상관분석 결과Benzo[a]pyrene 노출, 세포 생존력, p53, 인산화 p53, p21이 서로 높은 상관성을 보였다. 본 연구의 결과는p53 단백질의 축적이 benzo[a]pyrene 독성에 있어 매우 중요한 현상이며, 이는 선택적인 지표와 함께 p53이 benzo[a]pyrene과 같 은 PAH계열의 물질의 위해성 평가를 위한 민감한 생체 지표로써 개발될 수 있음을 시사한다.

Morphometric and Ultrastructural Change of Myelin-Associated Glycoprotein (MAG)-Immunoreactive Oligodendrocytes by Aging

조익현,박창현,이종환,배춘식,예상규,이법이,박승화,고기석,김진석,장병준,Cho, Ik-Hyun,Park, Chang-Hyun,Lee, Jong-Hwan,Bae, Chun-Sik,Ye, Sang-Kyu,Lee, Beob-Yi,Park, Seung-Hwa,Koh, Ki-Seok,Kim, Jin-Suk,Chang, Byung-Joon Korean Society of Electron Microscopy 2006 Applied microscopy Vol.36 No.2

정상적인 노화과정에 있어서 마이엘린연합당단백질(MAG)의 기능을 알아보고자 Sprague-Dawley 계통 흰쥐의 대뇌피질에서 MAG 면역양성반응세포(희소돌기아교세포)에 대한 형태계측학적 및 미세구조적인 분석을 시행하였다. 노화된 흰쥐의 대뇌피질(IV-VI)에서 MAG면역양성반응세포의 밀도는 정상 흰쥐에 비하여 유의하게 감소하였다. 그러나 medium과 dark 형의 희소돌기아교세포의 비율은 증가하였다. 노화된 흰쥐의 대뇌피질에서 MAG면역음성반응세포의 핵의 평균면적은 MAG면역양성반응세포의 핵의 평균면적보다 유의하게 감소하였다. MAG의 면역반응물은 노화된 흰쥐의 대뇌피질의 medium-dark형의 희소돌기아교세포의 세포질과 돌기에서 뚜렷이 감소하였고 dark형의 희소돌기아교세포에서는 거의 관찰되지 않았다. 이러한 결과는 노화에 의한 희소돌기아교세포와 마이엘린의 변성은 MAG의 감소와 관계가 있을 것으로 생각되며, 희소돌기아교세포와 마엘린 계통에서 MAG의 기능을 연구하는 기초자료로서 유용할 것으로 사료된다. To investigate the role of myelin-associated glycoprotein (MAG) in the normal aging process, aging-related morphometric and ultrastructural analyses of the MAG-positive (MAG-(+)) oligodendrocytes were carried out in the cerebral cortex of the Sprague-Dawley rats. In the aged rats, the density of MAG-(+) oligodendrocytes was significantly decreased in the cortical layer (IV-VI) compared with that of the adult rats. However, the percentage of medium and dark types of oligodendrocytes was significantly increased by aging. In the aged rats, the mean nuclear area of the MAG-(-) oligodendrocytes was interestingly reduced compared with that of MAG-(+) oligodendrocytes. In addition, MAG immunoreactive products were markedly decreased in the medium-dark type of oligodendroglial cytoplasm and processes, and were scarcely localized in the dark type of oligodendrocytes of the aged rats. These results suggest that degeneration of oligodendrocytes-myelin system by aging is associated with down regulation of MAG, and that may contribute to further understanding of the biology of MAG in the oligodendrocytes-myelin system.

Expression of Myelin-Associated Glycoprotein (MAG) in the Aged Rat Cerebrum

조익현,박창현,이종환,배춘식,예상규,이법이,박승화,고기석,김진석,장병준,Cho, Ik-Hyun,Park, Chang-Hyun,Lee, Jong-Hwan,Bae, Chun-Sik,Ye, Sang-Kyu,Lee, Beob-Yi,Park, Seung-Hwa,Koh, Ki-Seok,Kim, Jin-Suk,Chang, Byung-Joon Korean Society of Electron Microscopy 2006 Applied microscopy Vol.36 No.2

신경섬유의 수초화의 초기단계에 있어서 마이엘린의 형성에 중요한 역할을 한다고 알려져 있는 마이엘린연합 당단백질(MAG)이 정상적으로 노화된 흰쥐의 대뇌에서도 발현되는지를 알아보고자 하였다. 성숙흰쥐의 대뇌피질에서 MAG가 높은 농도로 발현되었으나 노화흰쥐의 대뇌피질에서는 유의하게 감소하였다. 대뇌에서 MAG면역양성반응 세포는 두 성숙흰쥐의 대뇌피질에서 주로 돌기를 가진 큰 세포였으며 노화흰쥐의 경우에는 주로 세포질과 돌기가 거의 없는 작고 둥근 세포였다. 성숙흰쥐의 백색질내 신경로에서 MAG면역양성 반응 세포는 많이 관찰되었으나 노화흰쥐에서는 거의 관찰되지 않았다. MAG면역반응은 galatocerebroside의 면역반응과 일치하였다. 이상의 결과로부터 노화에 의한 MAG 발현의 변화는 노화시에 나타나는 희소돌기아교세포와 마이엘린 퇴행성 변화와 관계가 있을 뿐만이 아니라 MAG는 노화시에 희소돌기아교세포의 기능 연구를 위한 적절한 marker로서 사용될 수 있음을 의미하며 앞으로 이에 대한 자세한 연구가 필요할 것으로 사료된다. Myelin-associated glycoprotein (MAG) has been known to have a crucial role to the formation of myelin sheath during initial stage of myelination. In the present study, we investigated the aging-related expressional changes of MAG in the rat cerebrum. MAG expression was markedly decreased in cerebral cortex by aging. In the adult rat cerebrum, MAG-positive rolls were process-bearing cells with large nucleus, and extensively distributed. However, in the aged rat brain, MAG-positive cells showed small and round morphology with little cytoplasm and few processes. MAG was co-expressed with galatocerebroside, but not with Iba-1, or GFAP. These results suggest that the expressional change of MAG-positive cells is associated with degeneration of oligodendrocyte-myelin system by aging, and that MAG is likely to be a reliable marker for the mature oligodendrocytes in the aged rat brain.

Comparison of the Antioxidant Activities of Nine Different Fruits in Human Plasma

정명희,고성희,최성원,예상규,조비농,김현숙 한국식품영양과학회 2005 Journal of medicinal food Vol.8 No.1

Oxidative stress in humans is associated with damage to DNA, protein and biological membrane. Oxidative stress, which often arises as a result of an imbalance in the human antioxidant status, has been implicated in aging and a number of human diseases such as cancer, atherosclerosis and rheumatoid arthritis. This study was performed to test the hypothesis that the consumption of fruit juices may improve antioxidant status in human plasma. Ten healthy men of age 25 to 26 were recruited for the study. After overnight-fasting, study subjects were supplied 150 ml of fruit juice, and blood was collected at 0, 30, 60, 90, and 120 minutes after consumption. After a one day wash-out period, subjects were fed with the next sample of fruit juice until all nine juices (pear, apple, orange, grape, peach, plum, kiwi, melon, water melon) had been evaluated. All juices were prepared from pure fruits ground in a home-style mixer. Dietary food records and anthropometrical measurements were used to evaluate the nutritional status of subjects. The antioxidant activity of fruit juices were estimated by measuring antioxidant status in the plasma using DCF fluorescence. Except for pear juice, eight kinds of juices exhibited potent antioxidant effects in human plasma. Within 30 minutes after consumption, orange, melon, grape, peach, plum, apple, and kiwi juices already effectively suppressed ROS generation. This radical scavenging effect of fruit juices was maintained for up to 90 minutes post consumption, but the relative DCF fluorescence had rebounded to near the initial levels at two hours post consumption in most samples tested. Interestingly however, grape juice continuously exerted the persistent antioxidant activity until two hours after supplementation. These results suggest that the consumption of fruits or fruit juices may reduce damage from oxidative stress, and that this effect may be a consequence of the antioxidant activity of fruits in scavenging the reactive oxygen species generated in human plasma. However, long-term studies with more subjects are needed to provide additional supportive evidence and better characterize the antioxidant properties of natural fruit juices.

Hypoxia activates the cyclin D1 promoter via the Jak2/STAT5b pathway in breast cancer cells

정연희,임은정,이문영,박종환,예상규,박의유,김상윤,Zheng Zhang,이광전,박동기,박태규,문원국,양영목 생화학분자생물학회 2005 Experimental and molecular medicine Vol.37 No.4

Hypoxia, a common consequence of solid tumor growth in breast cancer or other cancers, serves to propagate a cascade of molecular pathways which include angiogenesis, glycolysis, and various cell- cycle control proteins. As we have shown previously, hypoxia activates STAT5 (signal transducer and activator of transcription 5) and increases its binding activity to the GAS element in mammary epithelial cells. In this study we attempted to elucidate the mechanism by which cyclin D1 is regulated by the STAT5 protein under hypoxic conditions. Our data demonstrate that hypoxia (2% O2) or desferrioxamine (DFO) induces tyrosine and serine phosphorylation of STAT5 in human breast cancer cells (MCF-7) and mammary epithelial cells (HC11). Imunoprecipi-tation and subsequent Western analysis showed that activation of STAT5a or STAT5b under hypoxic conditions. Using a transfected COS-7 cell model system, we demonstrate that the activity of a cyclin D1 promoter-luciferase construct increased under hypoxic conditions or DFO treatment. The activity of the STAT5b/cyclin D1 promoter increased signifi-cantly by 12 h of hypoxia, whereas the activity of the STAT5a/cyclin D1 promoter was unaffected under hypoxic conditions. These increases in promoter activity are predominantly mediated by the Jak2/ STAT5b signaling pathway. We have shown by EMSA that hypoxia induces STAT5 to bind to the cyclin D1 promoter (GAS-1) in MCF-7 and HC11 cells. These data suggest that STAT5b may mediate the trans-criptional activation of cyclin D1 after hypoxic stimulation.

T Cell-Specific Knockout of STAT3 Ameliorates Dextran Sulfate Sodium-Induced Colitis by Reducing the Inflammatory Response

권순호,서은비,이송희,조정현,김성준,김상정,김항래,예상규 대한면역학회 2018 Immune Network Vol.18 No.4

Signal transducer and activator of transcription 3 (STAT3) has a crucial role in various autoimmune disorders including, inflammatory bowel disease (IBD). Our previous study demonstrated that STAT3 activation by IL-6 in colonic epithelial cells exacerbates experimental ulcerative colitis. Activated T lymphocytes are also found in ulcerative colitis patients with intestinal inflammation, but the role of STAT3 in T cells remains elusive. To determine the STAT3 function of T cells in intestinal inflammation, we generated T cell-specific STAT3 knockout (KO) mice and used dextran sulfate sodium (DSS) to induce colitis. In this study, we demonstrated that T cell-specific STAT3 deletion alleviated DSS-induced colitis in mice, resulting in reduced histological scores and myeloperoxidase (MPO) activity. Importantly, the population of T cells in the spleen and lymph nodes was significantly decreased in the control and DSS-induced groups of STAT3 KO mice. In addition, STAT3 deficiency in T cells markedly reduced the production of interferon (IFN)-γ, IL-6, and IL-17A, whereas IL-10 secretion was increased. Collectively, the results suggest that STAT3 in T cells may be a therapeutic target in ulcerative colitis by balancing the immune response through T cell homeostasis.

Anticancer Activity of Tubulosine through Suppression of Interleukin-6-Induced Janus Kinase 2/Signal Transducer and Activation of Transcription 3 Signaling

김병학,이은희,이옥진,박인철,박정열,예상규 한국유방암학회 2019 Journal of breast cancer Vol.22 No.3

Purpose: The chemical structure of tubulosine has been known since the mid-1960s. However, little is known about its biological and pharmacological functions. The aim of this study was to investigate the novel functions of tubulosine in cancer treatment, specifically in breast cancer. Methods: An Unpaired (Upd)-induced Drosophila cell line and interleukin (IL)-6-stimulated human breast cancer cell lines were used to investigate the biological and pharmacological activities of tubulosine in vitro. To investigate the activities of tubulosine, we performed molecular and cellular experiments such as Western blot and reverse transcription polymerase chain reaction analyses, immunoprecipitation and terminal deoxynucleotidyl transferase dUTP nick end labeling assays, and immunofluorescence staining using breast cancer cell lines. Results: Tubulosine exhibited anticancer activity in IL-6-stimulated human breast cancer cells. Moreover, tubulosine reduced the tyrosine phosphorylation level and transcriptional activity of signal transducer and activator of transcription (STAT) protein at 92E in Upd-induced Drosophila cells. Additionally, tubulosine suppressed IL-6-induced Janus kinase 2 (JAK2)/STAT3 signaling, resulting in decreased viability and induction of apoptotic cell death in breast cancer cells. Interestingly, inhibition of IL-6-induced JAK2/STAT3 signaling by tubulosine was associated with the blocking of IL-6 receptor (IL-6R) and glycoprotein 130 (gp130) binding. Conclusion: Tubulosine exhibits anticancer activity through functional inhibition of IL-6-induced JAK2/STAT3 signaling by targeting IL-6Rα/gp130 binding in breast cancer cells. These findings suggest that tubulosine may hold promise for the treatment of inflammation-associated cancers, including breast cancer.