OBJECTIVETo investigate the regulatory effect of the nonsteroidal anti-inflammatory drug NS398 on the expression of the RECK gene in the animal model of prostate cancer.

METHODSNude mouse models of prostate cancer were divided into an experimental and a control group, the former fed with NS398 at 0.1 mg/g per day for 10, 20 and 30 days, while latter left without medication. All the mice were killed at 30 days, the mRNA expressions of RECK and MMP-9 in the tumor tissues measured by RT-PCR, and the protein level of RECK evaluated by Western blot.

RESULTSBoth the mRNA and protein expressions of RECK were increased, while the level of MMP-9 decreased, in an obviously time-dependent manner in the experimental group as compared with the control.

CONCLUSIONNS398 obviously inhibits the pathogenesis and metastasis of prostate cancer, which may be attributed to its induction of the expression of the RECK gene and suppression of the expression of MMP-9.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; GPI-Linked Proteins ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Nude ; Nitrobenzenes ; pharmacology ; Sulfonamides ; pharmacology ; Tumor Cells, Cultured

OBJECTIVETo study the effects of NS398, a selective cyclooxygenase-2 (COX-2) inhibitor, on the transcription and translation of BCL-3 and its regulatory gene cyclin D1 in colon cancer cell line SW480.

METHODSHuman colon cancer cells SW480 were divided into two groups: SW480 cells in experimental group were treated with NS398 in different concentrations(25 micromol/L, 50 micromol/L, 100 micromol/L and 200 micromol/L) for 48 h or 72 h. SW480 cells in control group were treated with media which did not contain NS398. Then the expressions of BCL-3 and cyclin D1 were detected by RT-PCR, Western blot, and immunocytochemistry.

RESULTSAt 48 hours RT-PCR showed that BCL-3 mRNA and cyclin D1 mRNA decreased in a dose-dependent manner in the experimental group. However, there were no significant differences in the levels of BCL-3 protein and cyclin D1 protein between two groups (P>0.05). At 72 hours, BCL-3 protein and cyclin D1 protein also decreased in a dose-dependent manner in the experimental group. When the concentration of NS398 reached 100 micromol/L, the differences between the two groups in the expression of BCL-3 mRNA and protein became statistically significant (P<0.01). When the concentration of NS398 reached 50 micromol/L, the differences in the expression of cyclin D1 mRNA and protein were statistically significant (P<0.05).

CONCLUSIONSBCL-3 is expressed in colon cancer cell line SW480. COX-2 inhibitor can inhibit the expression of BCL-3 and cyclin D1 in a dose-dependent manner. NS398 may down-regulate the expression of cyclin D1 through BCL-3.

Cell Line, Tumor ; drug effects ; Colonic Neoplasms ; metabolism ; Cyclin D1 ; metabolism ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Humans ; Nitrobenzenes ; pharmacology ; Proto-Oncogene Proteins ; metabolism ; Sulfonamides ; pharmacology ; Transcription Factors ; metabolism

Retinoid resistance has limited clinical activity of retinoids as differentiation-inducing and apoptosis-inducing drugs. The present study was designed to investigate whether celecoxib (selective COX-2 inhibitor) has effects on cellular retinoid sensitivity of human colon cancer cell lines and its possible mechanism. Cell viability was measured by MTT assay. Apoptosis was detected by Annexin-V/PI staining and flow cytometry assay. PGE2 production was measured by ELISA assay. Expression of RARbeta was assayed by Western blotting. The results showed that celecoxib enhanced the inhibitory effect of ATRA in both COX-2 high-expressing HT-29 and COX-2 low-expressing SW480 cell lines. Further study showed the ATRA and celecoxib combination induced greater apoptosis, and the addition of PGE2 did not affect the number of apoptotic cells induced by the combination. Moreover, NS398 (another selective COX-2 inhibitor) did not affect the inhibitory effects of ATRA on both cell lines. Western blotting showed that the expression of RARbeta in HT-29 cell lines increased in celecoxib group and combination group. And the addition of PGE2 did not affect the expression of RARbeta induced by celecoxib either. In conclusion, celecoxib increased expression of RARbeta and cellular ATRA sensitivity through COX-2-independent mechanisms, which may provide a potential strategy for combination therapy.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Celecoxib ; Cell Line, Tumor ; Cell Survival ; drug effects ; Colonic Neoplasms ; metabolism ; pathology ; Cyclooxygenase 2 ; metabolism ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Dinoprostone ; metabolism ; Drug Synergism ; HT29 Cells ; Humans ; Nitrobenzenes ; pharmacology ; Pyrazoles ; pharmacology ; Receptors, Retinoic Acid ; metabolism ; Sulfonamides ; pharmacology ; Tretinoin ; pharmacology

OBJECTIVESTo observe the effects of NS-398 on proliferation of hepatic stellate cells (HSCs) in vitro, and to investigate the possible molecule mechanism.

METHODSHSCs were incubated with different concentrations of NS-398. The effects of NS-398 on cell proliferation was detected by MTT colormetric assay. The cell cycle of HSCs was analyzed by Flow Cytometry (FCM), cyclooxygenase-2 (COX-2) and proliferating cell nuclear antigen (PCNA) proteins in HSCs were detected by immunocytochemistry.

RESULTSAdministration of 20-160 micromol/L NS-398 significantly inhibited HSCs proliferation in dose-dependent manner compared with the control group (P less than 0.01). After treated with NS-398 at concentrations of 90, 120, and 150 micromol/L for 48 h, the number of HSCs in G(2)/M phase increased and the number of HSCs in G(0)/G(1) phase decreased (P less than 0.05); Incubated with 120 micromol/L NS-398 for 48 h, percentage of masculine cell of PCNA was 28.91%+/-0.11%, which was significantly lower than that of the control group (85.99%+/-0.13%) (P less than 0.01). Percentage of masculine cell of COX-2 was 13.80%+/-0.43%, which was not significantly different from that of the control group (14.07%+/-0.59%) (P more than 0.05).

CONCLUSIONSNS-398 could inhibit the proliferation of HSC-T6 and arrest HSCs in G2/M phase. Down-regulation of PCNA protein may partially accounted for the proliferation inhibition effect on HSCs induced by NS-398.

Animals ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; metabolism ; Cyclooxygenase Inhibitors ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Flow Cytometry ; Hepatic Stellate Cells ; drug effects ; metabolism ; Liver Cirrhosis ; pathology ; prevention & control ; Nitrobenzenes ; pharmacology ; Proliferating Cell Nuclear Antigen ; metabolism ; Rats ; Sulfonamides ; pharmacology

OBJECTIVETo examine modulations caused by cyclooxygenase-2 (COX-2) inhibitors on altered microenvironments and overbalanced neurotransmitters in pilocarpine-induced epileptic status rats and to investigate possible mechanisms.

METHODSCelecoxib (a COX-2 inhibitor) was administered 45 min prior to pilocarpine administration. The effects of COX-2 inhibitors on mIPSCs (miniature GABAergic inhibitory postsynaptic currents) of CA3 pyramidal cells in the hippocampus were recorded. Expressions of COX-2, c-Fos, newly generated neurons, and activated microgliosis were analyzed by immunohistochemistry, and expressions of alpha-subunit of gamma-amino butyric acid (GABA(A)) receptors and mitogen-activated protein kinase/extracellular signal-regulated protein kinase (MAPK/ERK) activity were detected by Western blotting.

RESULTSPretreatment with celecoxib showed protection against pilocarpine-induced seizures. Celecoxib prevented microglia activation in the hilus and inhibited the abnormal neurogenesis and astrogliosis in the hippocampus by inhibiting MAPK/ERK activity and c-Fos transcription. Celecoxib also up-regulated the expression of GABA(A) receptors. NS-398 (N-2-cyclohexyloxy-4-nitrophenyl-methanesulfonamide), another COX-2 inhibitor, enhanced the frequency and decay time of mIPSCs.

CONCLUSIONThe COX-2 inhibitor celecoxib decreased neuronal excitability and prevented epileptogenesis in pilocarpine-induced status epilepticus rats. Celecoxib regulates synaptic reorganization by inhibiting astrogliosis and ectopic neurogenesis by attenuating MAPK/ERK signal activity, mediated by a GABAergic mechanism.

Animals ; Blotting, Western ; Celecoxib ; Cyclooxygenase 2 ; metabolism ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Disease Models, Animal ; Fibrocystic Breast Disease ; metabolism ; Hippocampus ; drug effects ; enzymology ; pathology ; Immunohistochemistry ; MAP Kinase Signaling System ; drug effects ; Male ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Nitrobenzenes ; pharmacology ; Pilocarpine ; Proto-Oncogene Proteins c-fos ; metabolism ; Pyrazoles ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, GABA-A ; biosynthesis ; Status Epilepticus ; chemically induced ; enzymology ; pathology ; Sulfonamides ; pharmacology ; Synapses ; drug effects ; pathology

OBJECTIVETo investigate anticancer effect and molecular mechanism of N-[(Cyclohexyloxy)-4-nitrophenyl] methanesulfonamide on HepG2 cells in vitro.

METHODSHepG2 cells were treated with various concentrations (100, 200, 300, 400 micromol/L) of NS-398 [selective for cyclooxygenase 2 (COX-2) inhibition]. Cell growth was measured by MTT method, DNA fragmentation gel analysis was used to analyze the apoptosis cells, DNA ploidy and apoptotic cell percentage were examined by flow cytometry (FCM). PGE2 concentration was measured by radioimmunoassay method. The expressions of COX-2 were also examined by Western blot analysis.

RESULTSNS-398 inhibited HepG2 cell proliferation and induced apoptosis in a concentration-dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased and quiescent G1 phase was accumulated with NS-398 concentration increasing. The IC50 of 24 hours was 300 micromol/L. The release of PGE2 was significantly reduced in HepG2 cells with the values of NS-398 being (0.70 +/- 0.02), (0.48 +/- 0.02), (0.29 +/- 0.01) and (0.18 +/- 0.01) respectively, as compared with control group (0.03 +/- 0.01). NS-398 could inhibit the activity and expression of COX-2, with higher concentration, it can significantly down-regulate the expression of COX-2 (t = 3.736, 1.623, 1.810, 2.587, P < 0.01).

CONCLUSIONNS-398 might significantly inhibit the proliferation of HepG2 cells and induce apoptosis. The mechanisms were related with the accumulation of quiescent G1 phase and the inhibition of COX-2 activity.

Apoptosis ; drug effects ; Cell Line, Tumor ; metabolism ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; biosynthesis ; Humans ; Nitrobenzenes ; pharmacology ; Sulfonamides ; pharmacology

BACKGROUND/AIMS: Cyclooxygenase-2 (COX-2) inhibitors reportedly inhibit the growth of hepatocellular carcinoma (HCC) via caspase-dependent or caspase-independent apoptosis, which is due to COX-2 being associated with hepatocarcinogenesis. Survivin is highly expressed in most human cancers, but the mechanism regulating survivin expression remains unclear. We investigated the regulatory expression of survivin in selective-COX-2-inhibitor-induced growth inhibition of hepatoma cells. METHODS: After treatment with NS-398 (a selective COX-2 inhibitor) at various concentrations (10, 50, 100, 150, and 200 micrometer), the growth inhibition of Hep3B hepatoma cells was assessed by an MTT cell-viability assay, DNA fragmentation gel analysis, and flow cytometry. The expression of survivin transcript was analyzed by reverse-transcription polymerase chain reactions. RESULTS: NS-398 inhibited the growth of hepatoma cells by an amount dependent on the concentration and the time since treatment. Apoptotic DNA ladder and flow-cytometry shifting to the sub-G1 phase were revealed in NS-398-induced growth inhibition of hepatoma cells. NS-398 suppressed the expression of the survivin gene in a concentration- and time-dependent manner. CONCLUSIONS: Survivin was down-regulated in the growth inhibition of hepatoma cells induced by a selective COX-2 inhibitor, NS-398, in a concentration- and time-dependent manner. These results suggest the therapeutic inhibition of COX-2 via suppression of survivin in HCC.
Carcinoma, Hepatocellular/enzymology/*metabolism/pathology ; Cell Line, Tumor ; Cell Proliferation/drug effects ; Cyclooxygenase 2 Inhibitors/chemistry/*pharmacology ; G1 Phase ; Humans ; Liver Neoplasms/enzymology/*metabolism/pathology ; Microtubule-Associated Proteins/*antagonists & inhibitors/metabolism ; Neoplasm Proteins/*antagonists & inhibitors/metabolism ; Nitrobenzenes/chemistry/*pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Sulfonamides/chemistry/*pharmacology ; Time Factors

No abstract available.
Carcinoma, Hepatocellular/enzymology/*metabolism/pathology ; Cell Proliferation/drug effects ; Cyclooxygenase 2 Inhibitors/*pharmacology ; Cyclooxygenase Inhibitors/pharmacology ; Humans ; Liver Neoplasms/enzymology/*metabolism/pathology ; Microtubule-Associated Proteins/*antagonists & inhibitors/metabolism ; Neoplasm Proteins/*antagonists & inhibitors/metabolism ; Nitrobenzenes/pharmacology ; Sulfonamides/pharmacology

Cyclooxygenase 2 ; metabolism ; Cyclooxygenase Inhibitors ; pharmacology ; Hep G2 Cells ; Humans ; Nitrobenzenes ; pharmacology ; Radiation Tolerance ; drug effects ; Sulfonamides ; pharmacology

OBJECTIVESTo investigate the regulative effect of the nonsteroidal anti-inflammatory drug NS398 on the RECK gene in the prostate carcinoma strain DU145.

METHODSDU145 was treated with various concentrations of NS398 for 48 hours. The mRNA level was measured by RT PCR technique and the expression of the RECK protein determined by Western blot.

RESULTSThe mRNA level of the RECK gene was obviously higher, while the MMP9 level markedly lower in the treated group than in the control, and so was the expression of the RECK protein.

CONCLUSIONNS398 induces the expression of the RECK gene, which might be the mechanism of its anti-tumor effect.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Blotting, Western ; Cattle ; Cell Line, Tumor ; GPI-Linked Proteins ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Male ; Membrane Glycoproteins ; biosynthesis ; genetics ; Nitrobenzenes ; pharmacology ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sulfonamides ; pharmacology