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P
DPEP
1 2 2 1 2
1
2
Dipeptidase1 DPEP1 dipeptideglutathioneleukotrieneD4
GPI
DPEP1
HCC56
caveolin1 flotillin1
GPI CD59
DPEP1
GPI
P
Placenta specific miR-517a modulates gene expression in Jurkat
cells
Md Moksed Ali 1 , Xiaohui Song 1,2 , Osamu Ishibashi 1 , Kunio Kikuchi 1 ,
Tomoko Ishikawa 1 , Takami Takizawa 1 , Toshihiro Takizawa 1
1
Dept Mol Med & Anat, Nippon Medical School 2 Dept Pharm, Harbin Medical
University
[Objective] MicroRNAs miRNAs are noncoding RNAs that repress gene
expression posttranscriptionally. Recently we have found that human placenta
specific miRNAs are transferable to Jurkat cells human T cell lymphoblast
like cell line via exosomes manuscript in preparation. In this study, we
overexpressed placenta specific miR-517a in Jurkat cells and did microarray
analysis MA to identify its target mRNAs.
[Methods] MA was performed on Jurkat cells overexpressing miR-517a.
Validation of genes downregulated by miR-517a was carried out by realtime
PCR, Western blot, and 3'UTRluciferase reporter assay.
[Results] We found that cyclic GMP dependent protein kinase 1 (PRKG1), one
of 8 downregulated genes identified in MA, is a target of miR-517a in silico. MiR-
517a significantly downregulated the expression of PRKG1 at both the mRNA and
protein levels. Furthermore, miR-517a significantly decreased luciferase reporter
activity.
[Conclusion] We identified that PRKG1 is indeed a target of miR-517a in Jurkat
cells. These findings provide a mechanistic insight on the posttranscriptional
regulation by miRNAs of placentaderived exosomes in T cells.
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microRNA CpG
1 1,2 3 1
1
2 3
microRNA miRNA 22 RNA
19 miRNAs C19MC; miR-517a
Luo
et al. Biol Reprod 81: 717729, 2009 18
kb CG DNA
C19MC NoguerDance et al.
Hum Mol Genet 19: 35663582, 2010
BeWo JEG3
HTR8/SVneo C19MC
miRNA PCR
DNA PCR
CG
BeWoJEG3 C19MC HTR8/SVneo
BeWo, JEG3 C19MC
HTR8/SVneo
P
IgG IIb Fc
FcRIIb
1 2 1 3 1
1
2 3
HPEC
IgG
HPEC IIb Fc
FcRIIb IgG
J Immunol 175: 2331, 2005
FcRIIb RAB GTPasesRAB1B RAB3D
FcRIIb in vitro FcRIIb
GFP FcRIIb
pFCGR2B-GFP HUVEC
FcRIIb RAB1BRAB3D siRNA
FcRIIbsiRAB1B
siRAB3D HUVEC pFCGR2B-GFP
GFPFcRIIb GFPFcRIIb
HUVEC siRNA GFPFcRIIb
GFPFcRIIbHPEC
RAB1B RAB3D FcRIIb
P
Agerelated accumulation of nonheme ferric and ferrous iron in
mouse ovarian stroma visualized by sensitive iron histochemistries
Yoshiya Asano
Department of Neuroanatomy, Cell Biology and Histology, Hirosaki University
Graduate School of Medicine
Sensitive nonheme iron histochemistries, namely perfusionPerls and Turnbull
method, were applied to study the distribution and agerelated accumulation of
nonheme iron in mouse ovary. Microscopic studies revealed that nonheme ferric
iron is distributed predominantly in stromal tissue, especially in macrophages. By
contrast, the distribution of nonheme ferrous iron was restricted to a few ovoid
macrophages. Aged ovaries exhibited remarkable nonheme iron accumulation in
all stromal cells. Particularly, nonheme ferrous iron level was increased in stroma,
suggestive of increased levels of redoxactive iron, which can promote oxidative
stress. Moreover, intense localization of both nonheme ferric and ferrous iron was
observed in aggregated large stromal cells that were then characterized as ceroid
laden enlarged macrophages. Iron overload in adult mice resulted in nonheme
iron deposition in the stroma and generation of enlarged macrophages, suggesting
that iron accumulation induced morphological changes. Our data indicated that
nonheme iron accumulation in stromal tissue may be related to aging of the ovary
which due to the increasing of oxidative stress.
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CD34
CD141
5 μm CD34 CD141 DAB
CCD
CD34
CD141
CD34
CD34