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chGRP78 Suppresses Apoptosis by Phosphorylation of AKT1 at Serine 478 in Chicken Cells
Hoonsung Choi,Sun Keun Jung,Jeom Sun Kim,Kyung-Woon Kim,Keon Bong Oh,Hyeon Yang,Dong-Hoon Kim,Sung June Byun 한국동물생명공학회(구 한국동물번식학회) 2017 발생공학 국제심포지엄 및 학술대회 Vol.2017 No.10
GRP78 is typically expressed in mammalian cells and plays a prominent role in the maintenance of cell homeostasis and development. Additionally, chicken serum-mediated proliferation has regulated the chGRP78 to control prevented apoptosis in chicken cells, thus allowing the required chGRP78-mediated anti-apoptosis. However, the precise molecular mechanisms underlying the chGRP78-mediated apoptosis process remained to be defined. Previously, we reported the effects of chGRP78 on chicken embryo fibroblast (CEF) and DF-1 cell proliferation. Here we identify the AKT1 as a key target of chGRP78 in apoptosis mechanism. Using immunoprecipitation and peptide sequencing, chGRP78 binding sites were detected in AKT1-mediated protein. chGRP78 causes activation of AKT1, and chGRP78 knockdown cells show a decreased level of AKT1. Importantly, activation of AKT1 depletion reciprocally inhibits the apoptosis and promotes proliferation in chicken cells. Taken together, AKT1-mediated signaling pathway plays a critical role in chGRP78-stimulated fibroblast survival and anti-apoptosis, which, suggests an important implications of our findings for the maintenance of chicken fibroblast cells through the inhibition of apoptosis.
Isolation and Characterization of Spermatogonial Stem Cells in Chicken
Hoonsung Choi,Sun Keun Jung,Jeom Sun Kim,Kyung-Woon Kim,Keon Bong Oh,Hyeon Yang,Dong-Hoon Kim,Sung June Byun 한국동물생명공학회(구 한국동물번식학회) 2017 Reproductive & Developmental Biology(Supplement) Vol.41 No.2
Spermatogonial stem cells (SSCs) are expected to undergo self-renewal and differentiation into different body cell types in vitro, which are expected to serve as a powerful tool and resource for transgenic animal technology. We have established an efficient isolating for SSCs from 1-day-old chicken testicular cells. This present study is aimed to carry out the culture and characterization of chicken SSCs. Disassociation of chicken SSCs were performed using two-step enzymatic digestion of type 4 collagenase and trypsin and were seeded in a twelve-well plate. In primary culture was observed colony forming on day four post incubation. In the subsequent passages, chicken SSCs were positive for alkaline phosphatase activity and showed positive expression for the transcription factor, Oct 4. Immunoblotting analysis revealed that the chicken SSCs showed expression for the GRP78. Therefore, our findings may have implications for developmental biology and regenerative medicine by using chicken as the transgenic animal model.
Secular trends in the prognostic factors for papillary thyroid cancer
Choi, Hoonsung,Lim, Jung Ah,Ahn, Hwa Young,Cho, Sun Wook,Lee, Kyu Eun,Kim, Kyung Won,Yi, Ka Hee,Sung, Myung-Whun,Youn, Yeo-Kyu,Chung, June-Key,Park, Young Joo,Park, Do Joon,Cho, Bo Youn European Federation of Endocrine Societies 2014 European journal of endocrinology Vol.171 No.5
<P><B>Objective</B></P><P>With the recent increasing rates of screening for thyroid cancer, the cancers now tend to be smaller and less aggressive than those that are diagnosed when presented with symptoms, suggesting changes in the clinical validity of conventional prognostic factors for outcomes. We performed the retrospective study to identify the secular trends in the prognostic factors of thyroid cancer.</P><P><B>Methods</B></P><P>We used medical records of 3147 patients diagnosed with papillary thyroid cancer (PTC) at the Seoul National University Hospital Thyroid Cancer Clinic between 1962 and 2009.</P><P><B>Results</B></P><P>During the median 5.1-year follow-up, the overall recurrence rate was 13.3%, and male sex, tumor size, lymph node (LN) involvement, and extrathyroidal extension (ETE) were the significant prognostic factors for recurrence. Thyroid cancer-specific mortality was 1.4%, and the associated prognostic factors were older age, male sex, and LN involvement. For tumor recurrence, the hazard ratio (HR) for male sex decreased from 2.809 (95% CI, 1.497–5.269) in the pre-1989 period to 1.142 (95% CI, 0.736–1.772) in the post-1999 period. The pathologic characteristics, such as tumor size, LN involvement, and ETE, showed similar or increasing HRs over the time periods. For cancer-specific mortality, the HR for male sex decreased from 6.460 (95% CI, 1.714–24.348) in the pre-1990 period to 0.781 (95% CI, 0.083–7.379) in the post-1999 period.</P><P><B>Conclusion</B></P><P>The risk for poor outcomes in PTC associated with male sex decreased over time; in contrast, the risk associated with pathologic characteristics remained the same or increased over time. These trends might be associated with recent changes in the characteristics of patients with thyroid cancer.</P>
chGRP78 Is Required for the Phosphorylation of AKT1 in Chicken Cells
Hoonsung Choi,Sun Keun Jung,Jeom Sun Kim,Shamugam Sureshkumar,Hyeon Yang,Jae-Seok Woo,Gunsup Lee,Kyung-Woon Kim,Hwi-Cheul Lee,Keon Bong Oh,Sung June Byun 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06
Apoptosis occurs under various developmental, physiological, and pathological conditions, including normal chicken cell death. We have previously shown that chGRP78 is typically expressed in chicken cells and plays a prominent role in the maintenance of cell homeostasis and prevention of apoptosis. In addition, chicken serum-mediated proliferation regulates chGRP78 to prevent apoptosis in chicken cells via chGRP78- mediated anti-apoptosis. However, the precise molecular mechanisms underlying the chGRP78-mediated protection against apoptosis remain undefined. Here, we performed a comparative proteomic analysis of control and chGRP78-overexpressing DF1 cells to elucidate the cellular events that are directly or indirectly regulated by chGRP78. Using 2D gel based proteomics and bioinformatics prediction analysis, we detected chGRP78 binding sites in AKT1-regulated proteins. chGRP78 promoted AKT1 activation, and chGRP78 silencing decreased AKT1 levels. Therefore, our findings have important implications for the maintenance of chicken fibroblast cells via the inhibition of apoptosis.
Lee, Gunsup,Choi, Hoonsung,Sureshkumar, Shanmugam,Jung, Sun Keun,Kim, Jeom Sun,Oh, Keon Bong,Kim, Kyung-Woon,Yang, Hyeon,Kim, Dong-Hoon,Byun, Sung June Elsevier 2019 Research in veterinary science Vol.123 No.-
<P><B>Abstract</B></P> <P>Infectious bronchitis (IB) generated by the infectious bronchitis virus (IBV) causes economic difficulties for livestock farmers. The 3D8 single chain variable fragment (scFv) protein is a recombinant antibody with nuclease activity that shows antiviral effects against various DNA and RNA viruses in mice and chickens. In this experiment, 3D8 scFv G<SUB>2</SUB> transgenic chickens produced by crossing 3D8 scFv G<SUB>1</SUB> transgenic rooster and wild type hens were screened by genomic PCR and immunohistochemistry analysis. 3D8 scFv transgenic chickens, wild type sibling chickens, and SPF chickens were directly infected with IBV (5 chickens per group) and indirectly infected by airborne propagation (15 chickens per group). The relative IBV shedding titers were measured by quantitative real-time PCR using oropharyngeal and cloacal swabs on days 3 and 5 after intraocular infection. The viral load was significantly decreased in the 3D8 scFv transgenic chickens from the contact transmission group. Additionally, blood was collected from each group on day 17 post-infection. The ELISA results showed a marked reduction of the antibody titer against IBV in the 3D8 scFv transgenic chickens from the contact transmission group. These results suggest that the 3D8 scFv protein potentially inhibits infectious bronchitis virus transmission in chickens.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Produced G2 3D8 single chain variable fragment (scFv) transgenic chickens. </LI> <LI> 3D8 scFv transgenic chickens showed reduced infectious bronchitis viral shedding level in the contact transmission group. </LI> <LI> 3D8 scFv transgenic chickens were 40% lower than the response in the control groups in IBV serum antibody titer. </LI> </UL> </P>
Knockdown of the chPRDX3 Inhibits Cell Proliferation in Chicken Fibroblast Cells
Sun Keun Jung,Hoonsung Choi,Jeom Sun Kim,Shamugam Sureshkumar,Hyeon Yang,Jae-Seok Woo,Gunsup Lee,Hwi-Cheul Lee,Keon Bong Oh,Sung June Byun 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06
Chicken embryo fibroblast (CEF) cells are widely used in the study of embryology and differentiation. CEF and mammalian fibroblast cells differ in their requirements for defined nutrients in cell growth conditions. However, the molecular mechanism of the induced CEF proliferation is poorly understood. Our previous research has demonstrated that chicken GRP78 deficient promotes chicken cell apoptosis. In additionally, these studies identified the chicken Peroxiredoxin 3 protein (chPRDX3) in chicken cells by chicken serum-induced proliferation following two-dimensional (2D) and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) analyses. We had confirmed a relation between chPRDX3 upregulaion and cell proliferation in chicken embryo fibroblast cells. Our data demonstrated that the chPRDX3 significantly enhanced fibroblast proliferation. Moreover, knockdown of the chPRDX3 using siRNA decreased fibroblast proliferation and increased apoptosis. Therefore, the chPRDX3 is required for the cell proliferation in chicken fibroblast cells. Based on these results, we suggest that the chPRDX3 signaling pathway plays a critical role in chicken fibroblast survival. Therefore, our findings have important implications for the maintenance of chicken fibroblast cells.
Park, Jung-Ho,Lee, Jae-Woo,Choi, Hoonsung,Jung, Sun Keun,Kim, Jeom Sun,Kim, Kyung-Woon,Oh, Keon Bong,Yang, Hyeon,Byun, Sung June Elsevier 2018 Regulatory toxicology and pharmacology Vol.94 No.-
<P><B>Abstract</B></P> <P>Previously, <I>Escherichia coli</I> harboring the codon-optimized <I>3D8scFv</I> gene (<I>E. coli</I> 3D8scFv) was developed as a feed additive for use in preventing norovirus infection. Here, we evaluated whether the <I>3D8scFv</I> gene affects the colonization of <I>E coli</I> when <I>E. coli</I> 3D8scFv passes through the mouse gastrointestinal tract. To determine the colonization ability of <I>E. coli</I> 3D8scFv, <I>E. coli</I> cells with or without the <I>3D8scFv</I> gene were fed to mice. Total DNA was extracted from the animals’ stools, stomach, small intestine and colon. All samples were amplified using <I>3D8scFv</I> gene-specific primer sets. <I>E. coli</I> 3D8scFv begins to be excreted 1 h after feeding and that all <I>E. coli</I> 3D8scFv cells were excreted between 12 and 24 h after the last feeding of the cells. The previously measured gastrointestinal transit time of the mice was between 8 h and 22 h. The results of this study therefore show that <I>E. coli</I> 3D8scFv cannot colonize the gastrointestinal tracts of mice. In addition, if the purified 3D8 scFv protein is used as a feed additive, any associated <I>E. coli</I> 3D8scFv bacteria will not colonize the gastrointestinal tracts of the livestock. Thus, this feed additive meets the safety assessment criteria for the commercial use of bacteria.</P> <P><B>Highlights</B></P> <P> <UL> <LI> It is evaluated whether <I>E. coli</I> 3D8scFv colonizes in the gastrointestinal tracts of mice. </LI> <LI> Orally ingested <I>E. coli</I> 3D8scFv is excreted from mice without colonization of the gastrointestinal tract. </LI> <LI> Purified 3D8 scFv is suitable for a feed additive according to the concept of substantial equivalence. </LI> </UL> </P>