Apoptosis ; Calcineurin ; metabolism ; Chymotrypsin ; metabolism ; Humans ; Lysosomes ; enzymology ; Mitochondria ; metabolism ; pathology ; Mitochondrial Dynamics ; Neuroblastoma ; metabolism ; pathology

OBJECTIVETo study MMP-2, MMP-9, TIMP-2 and TIMP-1 expression, and their association to invasion and metastasis of neuroblastoma (NB).

METHODSThe staining status was compared of MMP-2, MMP-9, TIMP-2 and TIMP-1 in cryostat sections of tumor tissue in 35 NB patients by immunohistochemistry.

RESULTSStrong expression of MMP-2 was detected only in 2 patients with early stage NB (group A without metastasis), but in 9 and 10 respectively with advanced stage NB (group B with local metastasis and group C with distant metastasis) (compared to group A, P < 0.01). Strong MMP-9 staining was found in 4, 8 and 11 patients for group A, B and C patients (group A vs group C, P < 0.05). The expression of TIMP-2 was the strongest in 4 group A patients, but it decreased with progression of the disease. There was no statistical difference in TIMP-1 expression among the three groups of patients.

CONCLUSIONMMP-2, MMP-9 expression may be related to metastasis and progression of neuroblastoma, while TIMP-2 may have an inhibitory effect.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Lymphatic Metastasis ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Neuroblastoma ; enzymology ; metabolism ; secondary ; Retroperitoneal Neoplasms ; enzymology ; metabolism ; pathology ; Testicular Neoplasms ; enzymology ; metabolism ; secondary ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

Caffeine is one of the most widely consumed neuroactive drugs, coming mostly from everyday beverages such as coffee and tea. To investigate whether caffeine induces apoptosis in the central nervous system, 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, flow cytometric analysis, DNA fragmentation assay, and caspase-3 enzyme assay were performed on SK-N-MC human neuroblastoma cells. Cells treated with caffeine at concentrations as high as 10 mM exhibited several characteristics of apoptosis. In addition, caffeine was shown to increase the caspase-3 activity. These results suggest that high-dose of caffeine induces apoptosis in human neuroblastoma cells, probably by increasing the caspase-3 enzyme activity.
Apoptosis/*drug effects ; Caffeine/*toxicity ; Caspase 3 ; Caspases/metabolism ; Cell Cycle/drug effects ; Cell Survival/drug effects ; Central Nervous System/cytology/*drug effects ; DNA Fragmentation ; Humans ; Neuroblastoma/enzymology/pathology ; Tumor Cells, Cultured

Alcohol influences the neuroadaptation of brain cells where receptors and enzymes like protein kinase C (PKC) exist. Naltrexone acts on opioid receptors. However, other mechanisms of action remain unknown. We prepared SH-SY5Y neuroblastoma cells, and fed them with 150 mM ethanol for 72 h followed by treatment with naltrexone for 24 h. We performed microarray analysis and reverse transcriptase-polymerase chain reaction. Our results showed that PKC epsilon increased 1.90 times and showed an overall decreasing pattern as time increased. Phosphorylated ERK also increased 2.0 times according to the change of PKC epsilon. Integrin alpha7 increased 2.32 times and showed an increasing pattern as time increased. In conclusion, naltrexone influences PKC epsilon neuronal signaling system and endothelial adhesion molecule integrin alpha7 in addition to the well-known opioid system.
Antigens, CD/*metabolism ; Cell Line, Tumor ; Comparative Study ; DNA, Complementary/genetics ; Humans ; Integrin alpha Chains/*metabolism ; Naltrexone/*pharmacology ; *Neuroblastoma/enzymology/metabolism/pathology ; Oligonucleotide Array Sequence Analysis ; Protein Kinase C-epsilon/*metabolism ; Research Support, Non-U.S. Gov't ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors