• 제목/요약/키워드: Transduction

검색결과 1,264건 처리시간 0.033초

Effects of Adenoviral Gene Transduction on the Stemness of Human Bone Marrow Mesenchymal Stem Cells

  • Marasini, Subash;Chang, Da-Young;Jung, Jin-Hwa;Lee, Su-Jung;Cha, Hye Lim;Suh-Kim, Haeyoung;Kim, Sung-Soo
    • Molecules and Cells
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    • 제40권8호
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    • pp.598-605
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    • 2017
  • Human mesenchymal stem cells (MSCs) are currently being evaluated as a cell-based therapy for tissue injury and degenerative diseases. Recently, several methods have been suggested to further enhance the therapeutic functions of MSCs, including genetic modifications with tissue- and/or diseasespecific genes. The objective of this study was to examine the efficiency and stability of transduction using an adenoviral vector in human MSCs. Additionally, we aimed to assess the effects of transduction on the proliferation and multipotency of MSCs. The results indicate that MSCs can be transduced by adenoviruses in vitro, but high viral titers are necessary to achieve high efficiency. In addition, transduction at a higher multiplicity of infection (MOI) was associated with attenuated proliferation and senescence-like morphology. Furthermore, transduced MSCs showed a diminished capacity for adipogenic differentiation while retaining their potential to differentiate into osteocytes and chondrocytes. This work could contribute significantly to clinical trials of MSCs modified with therapeutic genes.

Induction of Signal Transduction Pathway Genes in Dendritic Cells by Lipopolysaccharides from Porphyromonas gingivalis and Escherichia coli

  • Jin, Ho-Kyeong;Lee, Young-Hwa;Jeong, So-Yeon;Na, Hee-Sam;Park, Hae-Ryoun;Chung, Jin
    • International Journal of Oral Biology
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    • 제35권3호
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    • pp.113-119
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    • 2010
  • Porphyromonas (P.) gingivalis lipopolysaccharide (Pg LPS) is the major pathogenic component of periodontal disease. In this study, we have attempted to determine the expression profiles of the signal transduction pathway genes induced by Pg LPS in comparison with Escherichia (E.) coli LPS (Ec LPS). DC2.4 cells were treated for two hours with $1\;{\mu}g/ml$ of Pg LPS or $0.5\;{\mu}g/ml$ of Ec LPS. The total RNA from these cells was then isolated and reverse-transcribed. Gene expression profiles were then analyzed with a signal transduction pathway finder GEArray Q series kit and significant changes in expression were confirmed by real-time PCR. The microarray results indicated that several genes, including Tnfrsf10b, Vcam1, Scyb9, Trim25, Klk6, and Stra6 were upregulated in the DC2.4 cells in response to Pg LPS treatment, but were downregulated or unaffected by Ec LPS. Realtime PCR revealed that the expression of Trim25, Scyb9 and Tnfrsf10b was increased over the untreated control. Notably, Trim25 and Tnfrsf10b were more strongly induced by Pg LPS than by Ec LPS. These results provide greater insight into the signal transduction pathways that are altered by P. gingivalis LPS.

Efficient Gene Delivery through the Human Transferrin Receptor of Fibroblast-like Synoviocytes Stimulated with bFGF: a Potential Target Receptor for Gene Transduction in Rheumatoid Arthritis

  • Kim, Hak-Jae;Joung, In-Sil;Nah, Seong-Su;Lee, Kyu-Hoon;KimKwon, Yun-Hee;Chung, Joo-Ho;Hong, Seung-Jae
    • Molecular & Cellular Toxicology
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    • 제3권2호
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    • pp.85-89
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    • 2007
  • Efficient gene delivery to specific tissues, such as inflammatory and cancerous tissues, is currently a major concern in disease treatment. The human transferrin receptor (hTR) has been detected in the synovium and fibroblast-like synoviocytes (FLS), which raises the possibility that expression of hTR is associated with the pathogenesis of rheumatoid arthritis (RA). To investigate whether the hTR is a useful target for gene transduction into the FLS of RA patients, recombinant adenoviruses with wildtype fiber (AdLac) and transferrin peptide-tagged fiber (Tf-AdLac) were used. The hTR expression level in FLS was notably increased by basic fibroblast growth factor (bFGF). Gene transduction to FLS was significantly higher by the hTR-targeted adenovirus than by the AdLac adenovirus, and treatment of the FLS with bFGF resulted in increased gene transduction by Tf-AdLac. Taken together, these data support Tf-AdLac as a new strategy for gene transduction in the treatment of RA patients.

근세포 분화에 관한 연구 : 근세포 분화에 있어서 Protein Kinase C (Studies on the Differentiation of Skeletal Muscle Cells in vitro:Protein Kinase C in the Differentiation of Skeletal Muscle Cells)

  • 최원철;김한도;김정락
    • 한국동물학회지
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    • 제34권2호
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    • pp.131-141
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    • 1991
  • TPA나 PDGF를 처리로 인한 Protein Kinase C의 신호전달은 힌산화에 의해 일어난다. 그렇지만, PKC에 의해 인산화 되어지는 targeting protein은 TAP나 PDGF 처리시에는 분자량이 서로 다른 단백질들이 인산화가 되어졌다. TPA처리한 myoblast에서 분자량 20,000의 단백질이 인산화되었다. PDGF처리한 세포에서는 분자량 40,000의 단백질이 인산화된 반면에 TPA처리로 인산화 되었던 분자량 20,000의 단백질은 탈인산화 되었다. 이러한 결과들은 TPA와 PDGF가 신호전달계의 활성에 있어서 다를 뿐만 아니라 그들은 장시간의 처리동안 PKC의 down regulation에 관계되어 짐을 암시한다. 그러나 PDGF는 TPA의 경우에서 보다 빠른 down regulation을 유도하였다. 면역세포 화학적인 연구에서 PKC의 동위효소인 PKC II는 세포질에, PKC III는 세포질과 인에 각각 분포하고 있었다. Myoblast에 있어서 PCK두가지 형태의 동위효소의 발현은 이들 동위효소들이 signal transduction이나 down regulation의 각기 다른 경로에 개입되어 진다는 것을 암시한다.

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Constitutive Activating Eel Luteinizing Hormone Receptors Induce Constitutively Signal Transduction and Inactivating Mutants Impair Biological Activity

  • Byambaragchaa, Munkhzaya;Choi, Seung-Hee;Kim, Dong-Wan;Min, Kwan-Sik
    • 한국발생생물학회지:발생과생식
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    • 제25권3호
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    • pp.133-143
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    • 2021
  • In contrast to the human lutropin receptor (hLHR) and rat LHR (rLHR), very few naturally occurring mutants in other mammalian species have been identified. The present study aimed to delineate the mechanism of signal transduction by three constitutively activating mutants (designated M410T, L469R, and D590Y) and two inactivating mutants (D383N and Y546F) of the eel LHR, known to be naturally occurring in human LHR transmembrane domains. The mutants were constructed and measured cyclic adenosine monophosphate (cAMP) accumulation via homogeneous time-resolved fluorescence assays in Chinese hamster ovary (CHO)-K1 cells. The activating mutant cells expressing eel LHR-M410T, L469R, and D590Y exhibited a 4.0-, 19.1-, and 7.8-fold increase in basal cAMP response without agonist treatment, respectively. However, inactivating mutant cells expressing D417N and Y558F did not completely impaired signal transduction. Specifically, signal transduction in the cells expressing activating mutant L469R was not occurred with a further ligand stimulation, showing that the maximal response exhibited approximately 53% of those of wild type receptor. Our results suggested that the constitutively activating mutants of the eel LHR consistently occurred without agonist treatment. These results provide important information of LHR function in fish and regulation with regard to mutations of highly conserved amino acids in glycoprotein hormone receptors.

Chemistry and Biology of Ras Farnesyltransferase

  • Cho, Kwang-Nym;Lee, Kee-In
    • Archives of Pharmacal Research
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    • 제25권6호
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    • pp.759-769
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    • 2002
  • Mutated forms of ras are found in many human tumors and the rate of incidence is significantly higher in colon and pancreatic cancers. The protein product from the ras oncogene is a small G-protein, $p21^{ras}{\;}(Ras)$ that is known to playa key role in the signal transduction cascade and cell differentiation and proliferation. Mutated Ras is unable to regulate itself and remains constantly activated, leading to uncontrolled cell growth. The function of Ras in signal transduction requires its location near the growth factor receptor at the cell membrane. However, Ras does not have a transmembrane domain. Ras requires farnesylation to increase its hydrophobicity and subsequent plasma membrane association for its transforming activity. This key post-translational modification is catalyzed by the enzyme Ras farnesyltransferase (FTase), which transfers a farnesyl group from farnesylpyrophosphate to the C-terminal cysteine of the Ras protein. The requirement has focused attention on FTase as a target for therapeutic intervention. Selective inhibition of FTase will prevent Ras protein from association with the plasma membrane, leading to a disruption of oncogenic Ras function.

Growth Inhibitory Patterns by Adenoviral p16 Transduction in HCC Cell Lines with Different pRB Status

  • Kim Keun-Cheol
    • 대한의생명과학회지
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    • 제11권4호
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    • pp.421-427
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    • 2005
  • To evaluate the diagnostic significance of p16 overexpression in human hepatocellular carcinoma (HCC), we analyzed p16 status and growth inhibitory patterns by p16 overexpression in HCC cell lines having different pRE status. SKHep1 and SNU449 cells show homozygous deletion of p16. The p16 gene in SNU398 cell is inactivated at posttranscription level. Adenovira1-p16 (Ad-p16) infection inhibits the cell growth in Hep3B, SNU398, and SNU449. Failure of growth inhibition in SKHepl results from the low transduction efficiency of adenovirus. The p16-mediated growth inhibition shows G 1 phase arrest in pRE-positive SNU449 but not in pRE-negative Hep3B. These results suggest that therapeutic efficacy of p16 gene might be considered on the transduction efficiency and the toxicity of adenoviral vector. Beside, growth inhibitory effect of p16 could be exerted through either pRE-dependent or -independent pathway.

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Characteristics of 14-3-3 Proteins and Their Role in Plant Immunity

  • Oh, Chang-Sik
    • The Plant Pathology Journal
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    • 제26권1호
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    • pp.1-7
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    • 2010
  • Phosphorylation is a major post-translational modification of proteins that regulate diverse signal transduction pathways in eukaryotic cells. 14-3-3 proteins are regulatory proteins that bind to target proteins in a phosphorylation-dependent manner and have been shown to play an important role in plant growth and development, primary metabolism, and signal transduction. Because phosphorylation plays a critical role in signal transduction pathways to trigger plant immunity, involvement of 14-3-3 proteins in plant immunity has been suggested for a long time. Recent studies have provided new evidence to support a role for 14-3-3 proteins in plant immunity. This review will briefly discuss general characteristics of 14-3-3 proteins and their involvement in plant immunity.

PHOTOMORPHOGENIC MUTANTS OF TOMATO

  • Kendrick, Richard E.
    • 한국식물학회:학술대회논문집
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    • 한국식물학회 1994년도 94 Symposium on Plant science September 10, 1994 Ewha Womans University, Seoul, Korea 94 식물학 심포지움 환경 스트레스와 식물의 반응
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    • pp.41-51
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    • 1994
  • Tomato (Lycopersicon esculentum Mill.) has been chosen as a model species for the study of hotomorphogenesis. The aurea (au) and yellow-green-2 (yg-2) mutants which are severely photochrome deficient appear to be phytochrome chromophore mutants. Mutants modified with respect to specific members of the phytochrome gene family: the far-red light-insensitive mutant (fri, for phytochrome A) and the temporarily red light-insensitive mutant (tri, for phytochrome B1) have been identified. Mutants that exhibit an exaggerated phytochrome response are putative transduction-chain mutants affecting an amplification step in phytochrome signal transduction. These mutants are being used to understand the complexities of juvenile anthocyanin in the hypocotyl during seedling de-etiolation.

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재조합 베큘로바이러스 벡터 시스템의 신 구축 (Novel Construction of Recombinant Baculovirus Vector System)

  • 사영희;홍성갑
    • 한국정보통신학회:학술대회논문집
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    • 한국정보통신학회 2012년도 추계학술대회
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    • pp.994-996
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    • 2012
  • 본 연구실에서는 새로운 베큘로바이러스 벡터 시스템을 구축하였다. 즉 본 벡터 시스템은 polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), protein transduction domain (PTD)을 코딩하는 유전자들로 구성 되어있다. 이렇게 새로이 제작된 베큘로바이러스 벡터 시스템과 대조군의 벡터 시스템과 효율과 발현율을 비교하였다. 그 결과 본 연구실에서 제작된 베큘로바이러스 벡터 시스템이 다른 대조군의 벡터 시스템에 비해 효과적임을 확인할 수 있었다.

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