PMID- 17285445
OWN - NLM
STAT- MEDLINE
DCOM- 20071009
LR  - 20181113
IS  - 0910-8327 (Print)
IS  - 0910-8327 (Linking)
VI  - 22
IP  - 1
DP  - 2007 Jan
TI  - RNA interference targeting embryonic myosin heavy chain isoform inhibited mRNA 
      expressions of phenotype markers in rabbit cultured vascular smooth muscle cells.
PG  - 41-7
AB  - To investigate whether the knockdown of SMemb gene expression induces phenotypic 
      modulation of vascular smooth muscle (VSM) cells toward a contractile type, we 
      constructed a siRNA targeting the 3' untranslated region (UTR) of SMemb gene 
      (SMemb-siRNA). The SMemb-siRNA was introduced into cultured rabbit VSM cells for 
      48 h at 37 degrees C by the lipofection method. The mRNA expressions were 
      estimated by comparative reverse transcription-polymerase chain reaction 
      (RT-PCR). SMemb-siRNA significantly decreased the ratio of SMemb to 
      glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA expression in a 
      dose-dependent manner (P < 0.01): 0 nM, 0.90 +/- 0.08; 100 nM, 0.43 +/- 0.07. 
      Immunofluorescence and immunoblot analyses demonstrated that SMemb-siRNA markedly 
      decreased SMemb protein expression to 56% +/- 7.8% (P < 0.01). Other MHC isoform 
      (SM1 and SM2) mRNA expressions were not changed. The relative mRNA expressions of 
      other phenotype markers (plasminogen activator inhibitor (PAI)-1 and beta-actin) 
      were significantly decreased by SMemb-siRNA to 71% +/- 7.5% and 61% +/- 7.5%, 
      respectively (P < 0.01). Expression of smooth muscle (SM) alpha-actin protein and 
      cell proliferation was not changed by SMemb-siRNA. Thus, SMemb gene might be 
      involved in the transcription of PAI-1 and beta-actin, but not involved in SM 
      alpha-actin and cell proliferation in cultured VSM.
FAU - Shimada, Seiji
AU  - Shimada S
AD  - First Department of Physiology, Unit of Physiological Science, School of 
      Medicine, University of the Ryukyus, 207 Uehara, Nishihara, Okinawa, 903-0215, 
      Japan.
FAU - Sunagawa, Masanori
AU  - Sunagawa M
FAU - Hanashiro, Kazuhiko
AU  - Hanashiro K
FAU - Nakamura, Mariko
AU  - Nakamura M
FAU - Kosugi, Tadayoshi
AU  - Kosugi T
LA  - eng
PT  - Journal Article
DEP - 20070126
PL  - Japan
TA  - Heart Vessels
JT  - Heart and vessels
JID - 8511258
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (Protein Isoforms)
RN  - EC 3.6.4.1 (Myosin Heavy Chains)
RN  - I223NX31W9 (Fluorescein-5-isothiocyanate)
SB  - IM
MH  - Animals
MH  - Cells, Cultured
MH  - Fluorescein-5-isothiocyanate
MH  - Fluorescent Dyes
MH  - Immunoblotting
MH  - Membrane Glycoproteins/metabolism
MH  - Muscle, Smooth, Vascular/*metabolism
MH  - Myocytes, Smooth Muscle/*metabolism
MH  - Myosin Heavy Chains/*metabolism
MH  - Phenotype
MH  - Protein Isoforms
MH  - RNA Interference/*physiology
MH  - Rabbits
MH  - Reverse Transcriptase Polymerase Chain Reaction
EDAT- 2007/02/08 09:00
MHDA- 2007/10/10 09:00
CRDT- 2007/02/08 09:00
PHST- 2005/12/12 00:00 [received]
PHST- 2006/05/27 00:00 [accepted]
PHST- 2007/02/08 09:00 [pubmed]
PHST- 2007/10/10 09:00 [medline]
PHST- 2007/02/08 09:00 [entrez]
AID - 10.1007/s00380-006-0929-x [doi]
PST - ppublish
SO  - Heart Vessels. 2007 Jan;22(1):41-7. doi: 10.1007/s00380-006-0929-x. Epub 2007 Jan 
      26.