OBJECTIVETo evaluate in vitro effect of abnormal savda munziq (ASMq) on the proliferation and apoptosis of human hypertrophic scar fibroblasts (HSFs).

METHODSHSFs were divided into six groups to receive different treatments as group A (blank control group), group B-E (ASMq in different concentration), and group F(5-Fu). Each group contains six specimens. The HSFs were cultured in vitro. After culture for 48 hours, the CCK8 test and flow cytometry methods were used to detect the proliferation, cell cycle and apoptosis.

RESULTSThe proliferation of HSFs in the B, C, D and E groups was inhibited at G2/M period, while it was inhibited at G0/S period in group F (P < 0.05). The inhibition effect of ASMq (0.1-1.0 mg/ml) on the fibroblasts enhanced in a concentration-dependent manner. Flow cytometry analysis with annexin V-FITC and PI staining confirmed the apoptotic. When HSFs were exposed to ASMq at 1.0 mg/ml (group E) for 48 h, the percentage of apoptotic cells increased to (43.7 +/- 2.58)%, which was significantly higher than that of blank control group (2.2 +/- 0.59)%. The induced apoptosis effect was also increased in a concentration-dependent manner.

CONCLUSIONASMq has a inhibitory effect on the proliferation and an enhancement effect on the apoptosis of fibroblast. ASMq could be used as an effective drug for treatment of hypertrophic scar.

Apoptosis ; Cell Cycle ; drug effects ; physiology ; Cell Division ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; pathology ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; In Vitro Techniques ; Medicine, East Asian Traditional

OBJECTIVETo compare the mechanisms of G(2)/M cycle arrest induced by topo IIalpha and IIbeta inhibitors in H460 cells.

METHODSThe inhibitory effects of XK469, adriamycin and etoposide on H460 cell growth were analyzed by MTT assay. The changes in cell cycle and expressions of cdc2, phos-cdc2 and 14-3-3sigma proteins induced by these 3 topo II inhibitors were detected by flow cytometry and Western blotting, respectively.

RESULTSBoth of the two types of topo II inhibitor resulted in dose-dependent G(2)/M phase arrest and growth inhibition of H460 cells, but XK469 failed to induce 14-3-3sigma protein expression as adriamycin and etoposide did.

CONCLUSIONTopo IIalpha and topo IIbeta inhibitors induce growth inhibition of H460 cells possibly through two different mechanisms, namely the 14-3-3sigma-dependent pathway and the 14-3-3sigma-independent pathway, but further functional inhibition test of 14-3-3sigma is needed to confirm this hypothesis.

Antigens, Neoplasm ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Cycle ; drug effects ; physiology ; Cell Division ; drug effects ; Cell Line, Tumor ; DNA Topoisomerases, Type II ; DNA-Binding Proteins ; antagonists & inhibitors ; G2 Phase ; Humans ; Lung Neoplasms ; pathology ; Quinoxalines ; pharmacology ; Topoisomerase II Inhibitors

OBJECTIVETo evaluate the effect of enamel matrix protein (EMP) on the attachment and proliferation of periodontal ligament cells (PDLC) on diseased cementum surfaces in vitro.

METHODSCementum chips were obtained from diseased roots exposed to periodontal pocket. Thirteen diseased root cementum chips were conditioned with EMP. Meanwhile, 13 diseased and 13 healthy cementum chips were treated with physiological saline as control. The growth and morphology of PDLC on the root surface were observed after 24 hours incubation by scanning electron microscope (SEM). PDLC attachment and proliferation were quantified using MTT assay at 16 or 72 hours.

RESULTSThe cells on EMP treated roots under SEM were growing robust like the cells on healthy roots. By contrast, the diseased cementum surface without conditioned with EMP was only partly covered with spindle-shaped cells, with filopodia appearing short and thin. MTT assay indicated that the number of adhered and proliferated cells on diseased cementum chips treated with EMP was significantly greater than that on diseased chips treated with saline (adhesion: 0.45 +/- 0.03 vs. 0.37 +/- 0.05, P < 0.05; proliferation: 0.71 +/- 0.02 vs. 0.55 +/- 0.08, P < 0.01), but less than that on healthy chips (adhesion: 0.45 +/- 0.03 vs. 0.67 +/- 0.08, P < 0.05; proliferation: 0.71 +/- 0.02 vs. 1.05 +/- 0.09, P < 0.05).

CONCLUSIONSIt was suggested that EMP could promote the growth of PDLC on the diseased root cementum surface.

Cell Adhesion ; drug effects ; Cell Division ; drug effects ; Cells, Cultured ; Dental Cementum ; physiology ; Dental Enamel Proteins ; pharmacology ; Humans ; Microscopy, Electron, Scanning ; Periodontal Ligament ; cytology ; Periodontitis ; pathology

Neuronal PC12 cells induced by nerve growth factor (NGF) have been considered to be postmitotic and lack the ability to divide. However, in this study, we not only detected DNA synthesis but also observed cell division in some morphologically differentiated neuronal PC12 cells bearing long neurites. More interestingly, in addition to the division of perikaryon, the neurites located on the division site of the cell membrane also divided into two parts and were allocated to the two daughter cells. These results demonstrate that the morphologically differentiated neuronal PC12 cells still retain the ability to divide. This is the first report that neuronal PC12 cells as well as their neurites can divide.
Animals ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; DNA Replication ; drug effects ; physiology ; Nerve Growth Factor ; pharmacology ; Neurites ; drug effects ; Neurons ; cytology ; PC12 Cells ; Rats

OBJECTIVETo investigate the neointima formation and the expression of monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-alpha (TNF-alpha) in cuff-induced vascular injury in mouse model, and to examine the effect of angiotensin II type 1 receptor (AT1) blocker, olmesartan, on MCP-1 and TNF-alpha expression and consequently vascular remodeling.

METHODSVascular injury was induced by polyethylene cuff-placement around the mouse femoral artery. Some mice were treated with AT1 receptor blocker, olmesartan, at the dose of 3 mg.kg(-1).day(-1) with an osmotic minipump. Neointima formation and the proliferation of vascular smooth muscle cells (VSMCs) were measured by morphometric analysis and bromodeoxyuridine (BrdU) incorporation. MCP-1 and TNF-alpha expression was detected by Western blot and immunohistochemical staining.

RESULTSWe observed neointima formation 14 days after cuff placement as well as VSMCs proliferation in the media and neointima. Cuff placement also induced MCP-1 and TNF-alpha expression in the media and neointima that the VSMCs specifically existed. Treatment of mice with olmesartan at a dose of 3 mg.kg(-1).day(-1), which did not influence systolic blood pressure, significantly decreased neointima formation and the proliferation of VSMCs. Olmesartan also inhibited MCP-1 and TNF-alpha expression in the injured arteries.

CONCLUSIONSOur results demonstrate that blockade of AT1 receptor inhibits MCP-1 and TNF-alpha expression and thereby improves vascular remodeling.

Analysis of Variance ; Animals ; Blotting, Western ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Chemokine CCL2 ; analysis ; Disease Models, Animal ; Imidazoles ; pharmacology ; Immunohistochemistry ; Male ; Mice ; Mice, Inbred C57BL ; Monocytes ; cytology ; drug effects ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Neovascularization, Physiologic ; drug effects ; physiology ; Olmesartan Medoxomil ; Probability ; Sensitivity and Specificity ; Tetrazoles ; pharmacology ; Tumor Necrosis Factor-alpha ; analysis ; drug effects ; Tunica Intima ; drug effects ; pathology ; Vascular Diseases ; physiopathology

OBJECTIVETo investigate the effect of basic fibroblast growth factor (bFGF) and hyaluronic acid (HA) on the proliferation of rabbit chondrocytes in vitro.

METHODSChondrocytes from the knee joints of New Zealand white rabbits were cultured. bFGF or HA or both were added into the culture medium respectively, and the proliferation of the chondrocytes was measured with MTT 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyl-tetra-zolium bromide. (MTT, Sigma, M2128).

RESULTSBasic fibroblast growth factor (10 ng/ml) with low concentration of fetal bovine serum in the culture medium promoted the proliferation of chondrocytes significantly, and this effect reached its maximum when concentration of bFGF reached 50 ng/ml. HA itself had no effect on the proliferation of chondrocytes. However, when bFGF was used in combination with HA, especially when the concentration of bFGF was 50-500 ng/ml and that of HA was 10-50 ng/ml, the effect on the proliferation of chondrocytes was much more than when bFGF or HA was used alone.

CONCLUSIONSbFGF can promote the proliferation of chondrocytes. HA, which has no effect on the proliferation of the cells, can maintain a normal growth of chondrocytes. When bFGF is used in combination with HA, more proliferation is obtained.

Analysis of Variance ; Animals ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Chondrocytes ; drug effects ; physiology ; Female ; Fibroblast Growth Factor 2 ; pharmacology ; Hyaluronic Acid ; pharmacology ; In Vitro Techniques ; Knee Joint ; cytology ; Male ; Probability ; Rabbits ; Sensitivity and Specificity

In order to explore the underlying mechanism of high effects and low toxicity of trichostatin A (TSA), the effect of TSA on growth inhibition, histone acetylation level and apoptosis in HL-60 cells and normal human peripheral blood mononuclear cells (NPBMNC) were examined using MTT method, immunocytochemistry technology, and Annexin-V-FITC/PI double staining flow cytometry. The results showed that TSA inhibited growth of HL-60 in time- and dose-dependent manners, and the IC(50) of 36 hours was 100 ng/ml. The apoptosis induction effect of TSA in HL-60 cells was also time- and dose-dependent. Besides, the dose of TSA showing significant apoptotic cytotoxicity in HL-60 cells did not demonstrate apparent apoptosis induction in NPBMNC within definite dose and time range. The histone acetylation level in HL-60 cells and NPBMNC both showed remarkable increase (P < 0.05) after incubated with 100 ng/ml TSA for 4 hours without statistical difference between them is detected (P > 0.05). It is concluded that TSA shows effects of definite and significant growth inhibition and apoptosis induction on HL-60 cells in time- and dose-dependent manners. TSA is able to selectively induce apoptosis in HL-60 cells with low toxicity in NPBMNC at the same time. The mechanism of this selectivity can not be ascribed to the differential regulation of histone acetylation level between HL-60 cells and NPBMNC.
Acetylation ; Apoptosis ; drug effects ; Cell Division ; drug effects ; DNA-Binding Proteins ; HL-60 Cells ; drug effects ; Histone Deacetylases ; physiology ; Histones ; metabolism ; Humans ; Hydroxamic Acids ; pharmacology ; RNA, Messenger ; analysis ; Telomerase ; genetics

The transwell chamber migration assay and the patch-clamp technique were used to investigate the volume-activated Cl(-) current (I(Cl.vol)) in migrated nasopharyngeal carcinoma cells (CNE-2Z). 47% hypotonic solution activated a ICl.vol in the migrated CNE-2Z cells. Compared with the control cells (non-migrated), the properties of this current and the sensitivity to Cl(-) channel blockers were changed. The current density in migrated CNE-2Z cells was higher than that in non-migrated cells. The current was almost completely inhibited by extracellular application of adenosine-5'-triphosphate (ATP, 10 mmol/L), 5-nitro-2-3-phenylpropylamino benzoic acid (NPPB, 100 mmol/L) and tamoxifen (30 mmol/L) in all voltage steps applied. The inhibition of NPPB and tamoxifen on the current was stronger in migrated cells than that in non-migrated cells. The permeability sequence of the four anions was Br(-)>Cl(-)> I (-)>Gluconate. The sequence was different from that of the non-migrated cells (I(-)> Br(-)> Cl(-)> Gluconate). The results suggest that volume-activated chloride channels may be involved in the CNE-2Z cell migration.
Carcinoma ; drug therapy ; metabolism ; pathology ; Cell Cycle ; drug effects ; physiology ; Cell Division ; drug effects ; Cell Movement ; drug effects ; Cell Size ; drug effects ; Chloride Channels ; antagonists & inhibitors ; metabolism ; physiology ; Chlorides ; metabolism ; Humans ; Nasopharyngeal Neoplasms ; drug therapy ; metabolism ; pathology ; Nitrobenzoates ; pharmacology ; Patch-Clamp Techniques ; Tamoxifen ; pharmacology ; Tumor Cells, Cultured

Antineoplastic Agents ; pharmacology ; Bile ; physiology ; Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Intrahepatic ; Celecoxib ; Cell Division ; drug effects ; Cell Line, Tumor ; Cholangiocarcinoma ; metabolism ; pathology ; Cyclooxygenase 2 ; Dinoprostone ; metabolism ; Humans ; Isoenzymes ; biosynthesis ; genetics ; Membrane Proteins ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; genetics ; Pyrazoles ; RNA, Messenger ; genetics ; Sphincterotomy, Transduodenal ; adverse effects ; Sulfonamides ; pharmacology ; Up-Regulation

BACKGROUNDExpression of polo-like kinase 1 (Plk1) is elevated in lung cancer and has been proposed as having prognostic value and related to resistance to chemotherapy and radiation. In addition, Plk1 has several functions in mitotic progression. In this study, the authors investigated the effect of Plk1 depletion on cell cycle progression and proliferation in A549 cells, a lung cancer cell line.

METHODSA recombinant plasmid containing antisense RNA targeting Plk1 (pcDNA3-Plk1) was transfected into A549 cells. Reverse transcription-polymerase chain reaction and Western blot were used to examine Plk1 gene expression. Cell proliferation was evaluated by direct cell counting and bromodeoxyuridine (BrdU) labelling. Cell cycle and apoptosis were examined by flow cytometry. Expression of alpha-tubulin was detected by immunofluorescence, and the inhibition rate (IR) by chemotherapeutic agents was determined by 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl-tetrazolium bromide assay.

RESULTSAfter transfection into A549 cells, pcDNA3-Plk1 reduced Plk1 mRNA by 46.75% for 24 hours and by 61.84% for 48 hours. Plk1 protein was significantly decreased simultaneously (P < 0.05). Abnormal morphological changes of cells and growth inhibition were observed in pcDNA3-Plk1 transfected groups. The BrdU labelling index was 25.59% 48 hours after transfection, which was significantly lower than that of the control groups (P < 0.05). Forty-eight hours after transfection, there showed absence of microtubule polymerization and spindle abnormalities in staining for alpha-tubulin. A549 cells showed a strong G2/M arrest and apoptosis 72 hours post transfection. IR of vinorelbine in pcDNA3-Plk1 transfected groups was significantly higher than that of the other groups (P < 0.05, respectively).

CONCLUSIONSPlk1 depletion interferes with spindle formation, induces cell cycle arrest and apoptosis, and consequently inhibits cell proliferation in A549 cells. Moreover, it sensitizes lung cancer cells to chemotherapy.

Apoptosis ; drug effects ; Bromodeoxyuridine ; metabolism ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; metabolism ; Cell Division ; drug effects ; Cell Line, Tumor ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; Protein Kinases ; genetics ; physiology ; Protein-Serine-Threonine Kinases ; Proto-Oncogene Proteins ; RNA, Antisense ; pharmacology ; Transfection ; Vinblastine ; analogs & derivatives ; pharmacology ; cdc25 Phosphatases ; metabolism