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Investigation of the DJ-1 Function in the Higher Mammals
Eun Young Kim,Lili Zhuang,Dong Ern Kim,Chi Sun Yun,Eun Ji Lee,Keun Jung Kim,Jae am Lim,Jun Jong Baek,Seung Bum Lim,Min Kyu Kim 한국동물생명공학회(구 한국동물번식학회) 2017 Reproductive & Developmental Biology(Supplement) Vol.41 No.2
The DJ-1 was found to be the first case that an oncogene also play a role for causative gene of neurodegenerative diseases. The DJ-1 is a multi-functional protein that play important role in tissue and participates in a number of intracellular signaling pathways. Especially, DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in canine. In the present study, we investigated the protective role of DJ-1 against induced oxidative stress, cell viability, cell proliferation assay, cellular apoptosis detection analysis, intracellular ROS were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions, whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions. The data indicate that canine DJ-1 protein enhances the cellular survival activity and promote cell survival activity under the oxidative stress by attenuates cellular apoptosis and ROS generation.
DJ-1 Gene Promote Cell Survival against Oxidative Stress in Canine
Eun Young Kim,Kang Sun Park,Bo Myeong Lee,Lili Zhuang,Dong Ern Kim,Eun Do Lee,Chi Sun Yun,Ji Hye Lee,Ju Lan Chun,Min Kyu Kim 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
The cancer and Parkinson's disease associated protein DJ-1 is multifunctional protein that involves in diverse cellular process. DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in novel organisms. In the present study, we investigated the protective role of DJ-1 against induced oxidative stress in canine cell line. On the basis of these experiments, canine DJ-1 overexpressing and null cell lines were established. The stable overexpression and down regulation of DJ-1 efficiency confirmed by the western blot analysis. Subsequently, the DJ-1 gene transfected cell lines and control cells were subjected to induced the oxidative stress, and then cell viability, cell proliferation assay, cellular apoptosis detection analysis (Annexin V and TUNEL assay), intracellular ROS and mitochondrial activity were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions induced by the rotenone and hydrogen peroxide (H2O2), whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions. Our data indicate that canine DJ-1 protein attenuates cellular apoptosis and ROS generation, enhances the cellular survival activity and promote mitochondrial function under the oxidative stress, likewise other mammalian cells. Importantly, DJ-1 overexpression may be an important part of a protective strategy as a sensor for oxidative stress.
Kwon, Dae-Jin,Kim, Dong-Hwan,Hwang, In-Sul,Kim, Dong-Ern,Kim, Hyung-Joo,Kim, Jang-Seong,Lee, Kichoon,Im, Gi-Sun,Lee, Jeong-Woong,Hwang, Seongsoo Springer International Publishing 2017 Transgenic research Vol.26 No.1
<P>Recent progress in genetic manipulation of pigs designated for xenotransplantation ha6s shown considerable promise on xenograft survival in primates. However, genetic modification of multiple genes in donor pigs by knock-out and knock-in technologies, aiming to enhance immunological tolerance against transplanted organs in the recipients, has not been evaluated for health issues of donor pigs. We produced transgenic Massachusetts General Hospital piglets by knocking-out the α-1,3-galactosyltransferase (GT) gene and by simultaneously knocking-in an expression cassette containing five different human genes including, DAF, CD39, TFPI, C1 inhibitor (C1-INH), and TNFAIP3 (A20) [GT<SUP>−(DAF/CD39/TFPI/C1-INH/TNFAIP3)/+</SUP>] that are connected by 2A peptide cleavage sequences to release individual proteins from a single translational product. All five individual protein products were successfully produced as determined by western blotting of umbilical cords from the newborn transgenic pigs. Although gross observation and histological examination revealed no significant pathological abnormality in transgenic piglets, hematological examination found that the transgenic piglets had abnormally low numbers of platelets and WBCs, including neutrophils, eosinophils, basophils, and lymphocytes. However, transgenic piglets had similar numbers of RBC and values of parameters related to RBC compared to the control littermate piglets. These data suggest that transgenic expression of those human genes in pigs impaired hematopoiesis except for erythropoiesis. In conclusion, our data suggest that transgenic expression of up to five different genes can be efficiently achieved and provide the basis for determining optimal dosages of transgene expression and combinations of the transgenes to warrant production of transgenic donor pigs without health issues.</P><P><B>Electronic supplementary material</B></P><P>The online version of this article (doi:10.1007/s11248-016-9979-8) contains supplementary material, which is available to authorized users.</P>