PMID- 16820933
OWN - NLM
STAT- MEDLINE
DCOM- 20070404
LR  - 20191210
IS  - 1107-3756 (Print)
IS  - 1107-3756 (Linking)
VI  - 18
IP  - 2
DP  - 2006 Aug
TI  - Real-time PCR as a tool for quantitative analysis of PI-PLCbeta1 gene expression 
      in myelodysplastic syndrome.
PG  - 267-71
AB  - Phosphoinositide-specific phospholipase C (PI-PLC) beta1 is a key enzyme in 
      nuclear signal transduction, and it is involved in many cellular processes, such 
      as proliferation and differentiation. In particular, the involvement of the 
      PI-PLCbeta1 gene in erythroid differentiation lead us to investigate this gene in 
      patients affected by high-risk myelodysplastic syndrome (MDS). By using 
      fluorescence in situ hybridization (FISH) analysis, we have previously evidenced 
      that, in MDS patients with normal GTG banding and a fatal outcome, the 
      PI-PLCbeta1 gene undergoes monoallelic and interstitial deletion. Real-time PCR 
      is characterized by high sensitivity, excellent precision and large dynamic 
      range, and has become the method of choice for quantitative gene expression 
      measurements. In the present study, we have performed a relative quantification 
      real-time polymerase chain reaction (PCR) analysis on all of the MDS patients 
      tested for FISH analysis. Furthermore, we have evaluated the expression of the 
      PI-PLCbeta1 gene on healthy donors and the HL60 cell line, which is useful for 
      testing the accuracy of the technology because of its low expression of 
      PI-PLCbeta1. To analyze and quantify the levels of the two different splicing 
      variants of PI-PLCbeta1 gene (1a and 1b), we have used a TaqMan isoform specific 
      probe. We have seen that all of the MDS patients have higher levels of the 
      PI-PLCbeta1 mRNA compared to the HL60 cell line as expected, but lower levels 
      compared to the healthy donors. Furthermore, MDS blasts always express higher 
      levels of PI-PLCbeta1b mRNA compared to PI-PLCbeta1a mRNA. Our data support the 
      contention that the deletion of the PI-PLCbeta1 gene is indeed responsible for a 
      reduced expression of the enzyme. In addition, the splicing isoform 1b, which is 
      only nuclear, seems to be somehow partially preserved compared to the 1a isoform, 
      which is nuclear and cytoplasmatic, hinting at a possible imbalance of the 
      nuclear versus cytoplasmatic PI-PLC signaling which, in turn, could affect the 
      cell cycle progression of MDS blasts.
FAU - Follo, Matilde Y
AU  - Follo MY
AD  - Department of Anatomical Sciences, Cellular Signalling Laboratory, University of 
      Bologna, 40126 Bologna, Italy.
FAU - Bosi, Costanza
AU  - Bosi C
FAU - Finelli, Carlo
AU  - Finelli C
FAU - Fiume, Roberta
AU  - Fiume R
FAU - Faenza, Irene
AU  - Faenza I
FAU - Ramazzotti, Giulia
AU  - Ramazzotti G
FAU - Gaboardi, Gian Carlo
AU  - Gaboardi GC
FAU - Manzoli, Lucia
AU  - Manzoli L
FAU - Cocco, Lucio
AU  - Cocco L
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PL  - Greece
TA  - Int J Mol Med
JT  - International journal of molecular medicine
JID - 9810955
RN  - 0 (Isoenzymes)
RN  - 0 (RNA, Messenger)
RN  - EC 3.1.4.11 (Phosphoinositide Phospholipase C)
RN  - EC 4.6.1.13 (Phosphatidylinositol Diacylglycerol-Lyase)
SB  - IM
MH  - Aged
MH  - Female
MH  - *Gene Expression Regulation
MH  - HL-60 Cells
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Isoenzymes/genetics/*metabolism
MH  - Male
MH  - Middle Aged
MH  - *Myelodysplastic Syndromes/enzymology/genetics
MH  - Phosphatidylinositol Diacylglycerol-Lyase/genetics/*metabolism
MH  - Phosphoinositide Phospholipase C
MH  - Polymerase Chain Reaction/*methods
MH  - RNA, Messenger/metabolism
EDAT- 2006/07/06 09:00
MHDA- 2007/04/05 09:00
CRDT- 2006/07/06 09:00
PHST- 2006/07/06 09:00 [pubmed]
PHST- 2007/04/05 09:00 [medline]
PHST- 2006/07/06 09:00 [entrez]
PST - ppublish
SO  - Int J Mol Med. 2006 Aug;18(2):267-71.