ABSTRACT
Objective To detect the expression of Hedgehog signaling pathway receptor Smoothened (Smo) protein in chronic myeloid leukemia (CML) CD34-positive (CD34+) cells, and to explore the role of Hedgehog signaling pathway in CML stem/progenitor cells. Methods Seventeen chronic phase (CP) and 6 advanced phase (AP) CML patients who were diagnosed in West China Hospital of Sichuan University from September 2010 to March 2011 and 10 People (healthy people and patients without hematologic malignances) as control were included in this study. The expression levels of Smo protein were detected by the protocol of indirect immunofluorescence labeling for cytometry and analyzed statistically. Results The mean of relative fluorescence intensity of Smo protein was 282.5±102.4, 188.8±55.4 and 354.0±297.9 in the CML-CP, CML-AD and control groups, respectively. The difference between CML-CP and CML-AD groups was statistically significant (P= 0.032). However, there were no significant differences between the CML-CP, CML-AD groups and control group (both P>0.05). The percentage of CD34+Smo+cells was (58.9±24.2)％, (42.6±17.6)％and (55.9±29.7) ％ in the CML-CP, CML-AD and control groups, respectively. There were still no significant differences between the CML-CP, CML-AD groups and control group (F= 0.950, P= 0.398). The mean of relative fluorescence intensity of Smo protein in the CML-CP patients with tyrosine kinase inhibitor (TKI) (9 cases) administered and without TKI (8 cases) were 282.3 ±122.6 and 282.6 ±82.4, respectively. There were no significant differences between the two groups(P=0.157). Conclusions Flow cytometry can qualitatively and quantitatively detect the expression level of Smo protein in CML CD34+cells. Smo expression is associated with stage of CML;TKI could not inhibit the activation of the Hedgehog signaling pathway in CML CD34+cells.
ABSTRACT
This study examined the effects of flow shear stress on the bio-capacity of the endothelial cells' induced from mesenchymal stem cells (MSCs). After cultivating the SD rat mesenchymal stem cells in vitro, we exposed them under different intensity of flow shear stress and induced these cells to endothelial cells. The variations of total anti-oxidation competence (T-AOC) and quantity of nitrogen monoxide (NO) were tested. The results showed that shear stress has an enhanced effect on the T-AOC and NO of endothelial cells induced from MSCs in an intensity-dependent manner. Flow shear stress could provide a protective action on the in vitro induction of endothelial cells, thus formulating a theoretical foundation for the therapeutics of ischemic heart diseases and vascular tissue engineering.
Subject(s)
Animals , Female , Male , Rats , Antioxidants , Metabolism , Bone Marrow Cells , Cell Biology , Physiology , Cell Differentiation , Physiology , Cells, Cultured , Endothelial Cells , Cell Biology , Metabolism , Mesenchymal Stem Cells , Cell Biology , Physiology , Oxidation-Reduction , Rats, Sprague-Dawley , Stress, MechanicalABSTRACT
<p><b>OBJECTIVE</b>To explore the relationship between NB4 cell apoptosis induced by tanshinone IIA (TanIIA) and the cell mitochondrial transmembrane potential (DeltaPsim).</p><p><b>METHODS</b>NB4 cells were treated with TanIIA, As(2)O(3), TanIIA plus 1.0 micro g/ml CsA and As(2)O(3) plus 1.0 micro g/ml CsA, respectively. Morphological changes were observed under light microscope and transmission electron microscope. The percentages of sub-G(1) cells and DeltaPsim of cells doublely stained with PI and Rh123 were assayed by flow cytometry.</p><p><b>RESULTS</b>The percentages of sub-G(1) cells after treatment with 1.0 micro g/ml and 2.0 micro g/ml TanIIA had no significant difference but was higher than that of 0.5 micro g/ml. After treatment with TanIIA, NB4 cells appeared the classical apoptotic morphology. The percentages of sub-G(1) cells were increased, while the DeltaPsim reduced (P < 0.01) and there was a linear correlation between them. The increment of sub-G(1) cell percentages and decrement of DeltaPsim induced by TanIIA were partly inhibited by CsA (P < 0.01).</p><p><b>CONCLUSIONS</b>TanIIA can induce NB4 cells apoptosis through opening the mitochondrial permeability transition pore and reducing DeltaPSgr;m, and this effect could be inhibited by CsA.</p>