Hakai acts as a coregulator of estrogen receptor alpha in breast cancer cells

Gong, Eun-Yeung,Park, Eunsook,Lee, Keesook Wiley (Blackwell Publishing) 2010 CANCER SCIENCE Vol.101 No.9

Expression of ectodermal neural cortex 1 and its association with actin during the ovulatory process in the rat.

Kim, Sun-Gyun,Jang, Soo-Jeong,Soh, Jaemog,Lee, Keesook,Park, Jin-Ki,Chang, Won-Kyong,Park, Eung-Woo,Chun, Sang-Young Association for the Study of Internal Secretions 2009 Endocrinology Vol.150 No.8

<P>Ectodermal neural cortex (ENC) 1, a member of the kelch family of genes, is an actin-binding protein and plays a pivotal role in neuronal and adipocyte differentiation. The present study was designed to examine the gonadotropin regulation and action of ENC1 during the ovulatory process in immature rats. The levels of ENC1 mRNA and protein were stimulated by LH/human chorionic gonadotropin (hCG) within 3 h both in vivo and in vitro. In situ hybridization analysis revealed that ENC1 mRNA was localized not only in theca/interstitial cells but also in granulosa cells of preovulatory follicles but not of growing follicles in pregnant mare's serum gonadotropin/hCG-treated ovaries. LH-induced ENC1 expression was suppressed by a high dose of protein kinase C inhibitor RO 31-8220 (10 microM) but not by low doses of RO 31-8220 (0.1-1.0 microM), suggesting the involvement of atypical protein kinase C. ENC1 was detected in both nucleus and cytoplasm that was increased by LH/hCG treatment. Both biochemical and morphological analysis revealed that LH/hCG treatment increased actin polymerization within 3 h in granulosa cells. Interestingly, ENC1 physically associated with actin and treatment with cytochalasin D, an actin-depolymerizing agent, abolished this association. Confocal microscopy further demonstrated the colocalization of ENC1 with filamentous actin (F-actin). The present study demonstrates that LH/hCG stimulates ENC1 expression and increases F-actin formation in granulosa cells. The present study further shows the physical association of ENC1 and F-actin, implicating the role of ENC1 in cytoskeletal reorganization during the differentiation of granulosa cells.</P>

ARR19 (androgen receptor corepressor of 19 kDa), an antisteroidogenic factor, is regulated by GATA-1 in testicular Leydig cells.

Qamar, Imteyaz,Park, Eunsook,Gong, Eun-Yeung,Lee, Hyun Joo,Lee, Keesook American Society for Biochemistry and Molecular Bi 2009 The Journal of biological chemistry Vol.284 No.27

<P>ARR19 (androgen receptor corepressor of 19 kDa), which encodes for a leucine-rich protein, is expressed abundantly in the testis. Further analyses revealed that ARR19 was expressed in Leydig cells, and its expression was differentially regulated during Leydig cell development. Adenovirus-mediated overexpression of ARR19 in Leydig cells inhibited testicular steroidogenesis, down-regulating the expression of steroidogenic enzymes, which suggests that ARR19 is an antisteroidogenic factor. Interestingly, cAMP/luteinizing hormone attenuated ARR19 expression in a fashion similar to that of GATA-1, which was previously reported to be down-regulated by cAMP. Sequence analysis of the Arr19 promoter revealed the presence of two putative GATA-1 binding motifs. Further analyses with 5' deletion and point mutants of putative GATA-1 binding motifs showed that these GATA-1 binding sites were critical for high promoter activity. CREB-binding protein coactivated GATA-1 and markedly increased the activity of the Arr19 promoter. Both GATA-1 and CREB-binding proteins occupied the GATA-1 motifs within the Arr19 promoter, which was repressed by cAMP treatment. Altogether, these findings demonstrate that ARR19 is the target gene of GATA-1 and suggest that ARR19 gene expression in testicular Leydig cells is regulated by luteinizing hormone/cAMP signaling via the control of GATA-1 expression, resulting in the control of testicular steroidogenesis.</P>

TNF-alpha as a Paracrine Factor in the Testis

Hong Cheol Yi,Park Jin Hee,Sub Ji Ho,Lee Keesook 한국발생생물학회 2004 한국발생생물학회 학술발표대회 Vol.18 No.-

Differential regulation of steroidogenic enzyme genes by TRα signaling in testicular Leydig cells.

Park, Eunsook,Kim, Yeawon,Lee, Hyun Joo,Lee, Keesook Endocrine Society 2014 Molecular endocrinology Vol.28 No.6

<P>Thyroid hormone signaling has long been implicated in mammalian testicular function, affecting steroidogenesis in testicular Leydig cells. However, its molecular mechanism is not well understood. Here, we investigated the molecular action of thyroid hormone receptor-α (TRα) on mouse testicular steroidogenesis. TRα/thyroid hormone (T3) signaling differentially affected the expression of steroidogenic enzyme genes, mainly regulating their promoter activity. TRα directly regulated the promoter activity of the cytochrome P450 17α-hydroxylase/C17-20 lyase gene, elevating its expression in the presence of T3. TRα also indirectly regulated the expression of steroidogenic enzyme genes, such as steroidogenic acute regulatory protein and 3β-hydroxysteroid dehydrogenase, by modulating the transactivation of Nur77 on steroidogenic enzyme gene promoters through protein-protein interaction. TRα enhanced Nur77 transactivation by excluding histone deacetylases from Nur77 in the absence of T3, whereas liganded TRα inhibited Nur77 transactivation, likely due to interfering with the recruitment of coactivator such as the steroid receptor coactivator-1 to Nur77. Together, these findings suggest a role of TRα/T3 in testicular steroidogenesis and may provide molecular mechanisms for the differential regulation of steroidogenic enzyme genes by thyroid hormone.</P>

NUP50 is necessary for the survival of primordial germ cells in mouse embryos

Park, Eunsook,Lee, Bobae,Clurman, Bruce E,Lee, Keesook Bioscientifica 2016 Reproduction Vol.151 No.1

<P>Nucleoporin 50 kDa (NUP50), a component of the nuclear pore complex, is highly expressed in male germ cells, but its role in germ cells is largely unknown. In this study, we analyzed the expression and function of NUP50 during the embryonic development of germ cells using NUP50-deficient mice. NUP50 was expressed in germ cells of both sexes at embryonic day 15.5 (E15.5), E13.5, and E12.5. In addition, NUP50 expression was also detected in primordial germ cells (PGCs) migrating into the genital ridges at E9.5. The gonads of <I>Nup50</I><I>−</I><I>/</I><I>−</I> embryos of both sexes contained few PGCs at both E11.5 and E12.5 and no developing germ cells at E15.5. The migratory PGCs in <I>Nup50</I><I>−</I><I>/</I><I>−</I> embryos at E9.5 showed increased apoptosis but a normal rate of proliferation, resulting in the progressive loss of germ cells at later stages. Taken together, these results suggest that NUP50 plays an essential role in the survival of PGCs during embryonic development.</P>

The expression of CKLFSF2B is regulated by GATA1 and CREB in the Leydig cells, which modulates testicular steroidogenesis

Kumar, Sudeep,Kang, Hana,Park, Eunsook,Park, Hee-Sae,Lee, Keesook Elsevier 2018 Biochimica et biophysica acta. Gene regulatory mec Vol.1861 No.12

<P><B>Abstract</B></P> <P>CKLFSF is a protein family that serves as a functional bridge between chemokines and members of the transmembrane 4 superfamily (TM4SF). In the course of evolution, CKLFSF2 has evolved as two isoforms, namely CKLFSF2A and CKLFSF2B, in mice. CKLFSF2A, also known as CMTM2A and ARR19, is expressed in the testis and is important for testicular steroidogenesis. CKLFSF2B is also known to be highly expressed in the testis. In the prepubertal stage, CKLFSF2B is expressed only in Leydig cells, but it is highly expressed in haploid germ cells and Leydig cells in adult testis. CKLFSF2B is naturally processed inside the cell at its C-terminus to yield smaller proteins compared to its theoretical size of ≈25 kDa. The <I>Cklfsf2b</I> gene is regulated by GATA-1 and CREB protein, binding to their respective binding elements present in the 2-kb upstream promoter sequence. In addition, the overexpression of CKLFSF2B inhibited the activity of the Nur77 promoter, which consequently represses the promoter activity of Nur77-target steroidogenic genes such as P450c17, 3β-HSD, and StAR in MA-10 Leydig cells. Adenovirus-mediated overexpression of CKLFSF2B in primary Leydig cells isolated from adult mice shows a repression of steroidogenic gene expression and consequently testosterone production. Moreover, intratesticular injection of CKLFSF2B-expressing adenovirus in adult mice clearly had a repressive effect compared to the control injected with only GFP-expressing adenovirus. Altogether, these findings suggest that CKLFSF2B might be involved in the development and function of Leydig cells and regulate testicular testosterone production by fine-tuning the expression of steroidogenic genes.</P> <P><B>Highlights</B></P> <P> <UL> <LI> CKLFSF2B is expressed in the Leydig cells and processed at C-terminus naturally. </LI> <LI> The <I>Cklfsf2b</I> gene expression is regulated by GATA-1 and CREB protein. </LI> <LI> CKLFSF2B modulates testicular steroidogenesis by regulating Nur77 expression. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Estrogen receptor-related receptor γ regulates testicular steroidogenesis through direct and indirect regulation of steroidogenic gene expression

Park, Eunsook,Kumar, Sudeep,Lee, Bobae,Kim, Kyung-Jin,Seo, Jeong-Eun,Choi, Hueng-Sik,Lee, Keesook Elsevier 2017 Molecular and cellular endocrinology Vol.452 No.-

<P><B>Abstract</B></P> <P>Biosynthesis of testosterone, which mainly occurs in testicular Leydig cells, is controlled by steroidogenic proteins, such as StAR and P450c17. Although estrogen-related receptor gamma (ERRγ), an orphan nuclear receptor, is expressed in the testis, its role is not well understood. In this study, we investigated the expression of ERRγ in Leydig cells and its molecular action on testicular steroidogenesis. ERRγ is expressed in mouse Leydig cells from pre-pubertal stages. ERRγ overexpression in primary Leydig cells elevated the production of testosterone with a marked increase of P450c17 expression at both mRNA and protein levels, albeit decreased expression of StAR. Promoter-reporter analyses showed that ERRγ directly regulated the P450c17 promoter. Further deletion mutant analyses of the P450c17 promoter revealed that ERRγ activated expression of the P450c17 gene by binding to an ERRγ response element within the P450c17 promoter. Meanwhile, ERRγ suppressed cAMP-induced activation of the StAR promoter, which was likely due to ERRγ-mediated inhibition of the transcriptional activity of Nur77, which is induced by cAMP and regulates StAR gene expression in Leydig cells. Interestingly, ERRγ coexpression also decreased the protein level of Nur77, which occurred through proteasomal degradation, suggesting ERRγ-mediated regulation of steroidogenesis at another level. Taken together, these findings suggest that ERRγ regulates testicular steroidogenesis, both directly controlling and indirectly fine-tuning the expression of steroidogenic genes.</P> <P><B>Highlights</B></P> <P> <UL> <LI> ERRγ elevates testosterone production in testicular Leydig cells. </LI> <LI> ERRγ directly regulates the P450c17 promoter. </LI> <LI> ERRγ inhibits Nur77 transactivation and reduces the Nur77 protein stability; thus regulates cAMP/Nur77 regulated promoters. </LI> </UL> </P>

SAR Based Design of Nicotinamides as a Novel Class of Androgen Receptor Antagonists for Prostate Cancer

Yang, Su Hui,Song, Chin-Hee,Van, Hue Thi My,Park, Eunsook,Khadka, Daulat Bikram,Gong, Eun-Yeung,Lee, Keesook,Cho, Won-Jea American Chemical Society 2013 Journal of medicinal chemistry Vol.56 No.8

<P>Molecular knowledge of pure antagonism and systematic SAR study offered a direction for structural optimization of DIMN to provide nicotinamides as a novel series of AR antagonists. Nicotinamides with extended linear scaffold bearing sterically bulky alkoxy groups on isoquinoline end were synthesized for H12 displacement. AR binding affinity and molecular basis of antiandrogenic effect establish the optimized derivatives, <B>7au</B> and <B>7bb</B>, as promising candidates of second generation AR antagonists for advanced prostate cancer.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jmcmar/2013/jmcmar.2013.56.issue-8/jm3014103/production/images/medium/jm-2012-014103_0011.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/jm3014103'>ACS Electronic Supporting Info</A></P>

Overexpression of hepatocyte nuclear factor-3α induces apoptosis through the upregulation and accumulation of cytoplasmic p53 in prostate cancer cells

Lee, Hyun Joo,Chattopadhyay, Soma,Yoon, Wan-Hee,Bahk, Jong Yoon,Kim, Tae-Hyoung,Kang, Hyung Sik,Lee, Keesook Wiley Subscription Services, Inc., A Wiley Company 2010 The Prostate Vol.70 No.4

<B>BACKGROUND</B><P>Hepatocyte nuclear factor-3α (HNF-3α) has been known to act as a repressor in the pathogenesis of many cancers. Herein, we investigated the effect of HNF-3α overexpression in prostate cancer cells.</P><B>METHODS</B><P>HNF-3α was overexpressed in prostate cancer cells using an adenovirus recombinant expressing wild-type HNF-3α. The apoptosis of prostate cancer cells was determined by TUNEL, FACS, and caspase activity analyses.</P><B>RESULTS</B><P>Adenovirus-mediated overexpression of HNF-3α caused cell death in prostate cancer cells as assessed by changes in cellular and nuclear morphology, TUNEL analysis, and caspase activations. Furthermore, FACS analysis showed an increased sub-G1 phase of cell cycle as well as the G2/M phase with a corresponding decrease in S phases. HNF-3α overexpression caused the upregulation of p53 protein and its accumulation, together with HNF-3α, in the cytoplasm. It also causes Bax protein to localize to the mitochondria-enriched fraction. These findings suggest that multiple apoptotic pathways seem to be involved in the HNF-3α-induced cell death: pathways involving the accumulation of p53 protein in the cytoplasm and subsequent cytochrome c release, and other pathways involving death receptor signaling and caspase-8 activation.</P><B>CONCLUSIONS</B><P>The results of the current study suggest a novel function of HNF-3α as a killer of malignant prostate cancer cells, which reveals HNF-3α as a promising therapeutic molecule for prostate cancers. Prostate 70: 353–361, 2010. © 2009 Wiley-Liss, Inc.</P>