Cell Line, Tumor ; Humans ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 9 ; analysis ; Melanoma ; enzymology ; Skin Neoplasms ; enzymology

Child ; Ear Auricle ; pathology ; Ear Neoplasms ; Female ; Humans ; Neurilemmoma

Adult ; Aged ; Female ; Humans ; Joint Instability ; complications ; diagnosis ; physiopathology ; surgery ; Lumbar Vertebrae ; pathology ; physiopathology ; Male ; Middle Aged ; Spinal Canal ; pathology ; physiopathology ; Spinal Stenosis ; complications ; diagnosis ; physiopathology ; surgery

BACKGROUNDTransforming growth factor beta (TGF-beta) plays an essential role in the regulation of normal physiologic processes of cells. TGF-beta has been shown to regulate several mitogen-activated protein kinases (MAPK) pathways in several epithelial cells. However, the effects of TGF-beta on soft tissue sarcoma are seldom reported. Our previous studies suggested that there should be some other signal transduction pathways besides Smads, which are important to regulate the growth of human embryonal rhabdomyosarcoma (RMS) cells. In the present study, we examined the expression and functional relations of extracellular signal-regulated kinase 2 (ERK2) and Smad4 in human RMS tissue and a RMS cell line, RD.

METHODSRD cells and normal human primary skeletal myoblasts (Mb) were treated with TGF-beta1 to establish the expression profile of ERK2 at the mRNA and protein levels detected by RT-PCR and immunofluorescence. Immunohistochemistry was used to detect the expression of ERK2 and Smad4 in 50 tissue specimens of human RMS and 23 specimens of normal skeletal muscles. Follow-up of specimens was performed 6 months to 70 months later.

RESULTSRD cells and human RMS tissues showed the higher expression of ERK2 and Smad4 than the normal control, either the protein level or the mRNA level. And, exogenous TGF-beta1 stimulation can lead to higher expression of ERK2 and its nuclear translocation, so TGF-beta1 can also activated MAPK (ERK2) pathway, resulting in a sustained activation of ERK2 for at least 2 hours. Immunohistochemistry analysis, however, showed that there was no correlation between ERK2 and Smad4 protein. The overexpression of ERK2 and Smad4 had no indicative effects on histological subtypes, histological grading, gender, age, and prognosis.

CONCLUSIONSIn RMS, signaling of TGF-beta1 from cell surface to nucleus can also be directed through the MAPK (ERK2) pathway besides the TGF-beta1/Smads pathway. The activation of ERK2 by TGF-beta1 may be Smad4 independent. Moreover, there may be some other tanglesome relationships between the TGF-beta1/Smads pathway and the MAPK pathway which takes part in the development, invasion and metastasis of tumor cells.

Cells, Cultured ; Humans ; Mitogen-Activated Protein Kinase 1 ; physiology ; Muscle, Skeletal ; metabolism ; RNA, Messenger ; analysis ; Rhabdomyosarcoma ; metabolism ; Signal Transduction ; Smad4 Protein ; physiology ; Transforming Growth Factor beta1 ; pharmacology

BACKGROUNDTransforming growth factor-beta (TGF-beta) can inhibit the growth of most epithelial and endothelial cells. The growth regulative role of TGF-beta on soft tissue sarcoma was seldom reported. Here we examined TGF-beta1 effects on the growth of human rhabdomyosarcoma cell RD and searched the relative molecular mechanism.

METHODSThe viability of RD was examined by [(3)H]-thymidine incorporation and [3-(4,5-dimethylthiazol-z-yl)-2,5-diphenyl tetrazolium bromide] (MTT) assay. RD cell cycle was analysed by flow cytometry. The protein and mRNA of cell cycle regulative factors in RD were detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. The kinase activity of cdk2 or cdk4 was examined by immunoprecipitation and kinase assay. Immunofluorescent staining was used to detect the location of cell cycle regulative factors in RD by laser scanning confocal microscope.

RESULTSTGF-beta1 inhibits RD proliferation by G1-arrest in cell cycle progression. TGF-beta1 can prominently up-regulate P27 of RD, then augment P27 to bind cyclinE-cdk2 complexes, which effectively suppress cdk2 kinase activity. P21 increased and c-myc decreased in RD due to TGF-beta1. Both P15 and cdk4 have not been involved in the growth inhibitory event. TGF-beta1 treatment induced P27 to congregate around nucleus. P21 pervaded from nucleus to both nucleus and cytoplasm by TGF-beta1 treatment.

CONCLUSIONTGF-beta1 inhibits the proliferation of human rhabdomyosarcoma cell line RD and induces RD G1-arrest. This course is accomplished by TGF-beta1 up-regulating P27 to suppress cdk2 kinase activity. The induction of P21 and down-regulation of C-myc might participate in the growth-arrest event.

Cell Cycle Proteins ; analysis ; genetics ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; G1 Phase ; drug effects ; Genes, myc ; Humans ; RNA, Messenger ; analysis ; Rhabdomyosarcoma ; pathology ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1 ; Tumor Suppressor Proteins ; analysis ; genetics

OBJECTIVETo study the regulatory effect of TGF-beta1 on growth of rhabdomyosarcoma RD cell line.

METHODSAfter various durations of TGF-beta1 treatment, the viability of RD cell line was examined by growth rate measurement, MTT assay and (3)H-thymidine incorporation. The cell cycle was analyzed by flow cytometry. Immunofluorescent staining was used to localize p15, p21 and p27 in RD cell line under laser scanning confocal microscope. The protein and mRNA of p15, p21 and p27 in RD cell line were detected by western blot and reverse transcriptase-polymerase chain reaction respectively.

RESULTSThe viability of RD cell line treated with TGF-beta1 was obviously decreased. RD cell line was arrested in G(1) phase by TGF-beta1. There was increased expression of p21 and p27 in RD cell line with TGF-beta1 treatment at protein and mRNA levels. The expression of p21 in RD cell line was seen in both nucleus and cytoplasm after 24 hours of TGF-beta1 treatment. The expression of p15 showed no obvious changes upon TGF-beta1 treatment.

CONCLUSIONSTGF-beta1 inhibits growth of RD cell line and induces G(1)-arrest. It up-regulates protein and mRNA of p21 and p27 and shows no obvious influence on p15 expression. The growth arrest of RD cell line may result from the up-regulation of p21 and p27 by TGF-beta1.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p15 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p27 ; biosynthesis ; genetics ; G1 Phase ; drug effects ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Rhabdomyosarcoma ; pathology ; Transforming Growth Factor beta1 ; pharmacology

OBJECTIVETo study the effects of TGF-beta/Smad signaling on the growth and apoptosis of human rhabdomyosarcoma cell line RD.

METHODSBiosynthesized short hairpin RNA (shRNA) was transfected into RD cells by cation liposome vector to suppress Smad4 expression. The mRNA and protein expression of Smad4 in RD after shRNA-transfection were examined by RT-PCR and Western blot respectively. Immunofluorescent staining was used to detect the location of Smad2/3 in RD by laser scanning confocal microscopy. The viability of RD cells was examined by MTT method and (3)H-thymidine incorporation assay. The apoptosis of RD was examined by flow cytometry analysis and fluorescent staining.

RESULTSThe expression of mRNA and protein of Smad4 in RD were effectively suppressed by shRNA interference. Such suppression effectively interrupted the endogenous TGF-beta/Smad signaling and consequently blocked the translocation of Smad2/3. The interruption of endogenous TGF-beta/Smad signaling not only inhibited the growth of RD but also induced apoptosis of RD. Exogenous TGF-beta1 inhibited the growth of RD but did not influence the apoptosis of RD.

CONCLUSIONshRNA interference can effectively interrupt the TGF-beta/Smad signaling by suppressing the expression of Smad4. TGF-beta/Smad signaling subtly regulates the growth and apoptosis of RD.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Cell Proliferation ; drug effects ; Cytoplasm ; metabolism ; Humans ; Protein Transport ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; pharmacology ; Rhabdomyosarcoma ; genetics ; metabolism ; pathology ; Signal Transduction ; Smad2 Protein ; metabolism ; Smad3 Protein ; metabolism ; Smad4 Protein ; biosynthesis ; genetics ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

OBJECTIVETo observe the histologic changes of the cervical muscles and intervertebral discs caused by dynamic dysequilibrium of frontally cervical muscles in rabbits.

METHODSThirty healthy rabbits with an average age of two years, half males and half females, the mean of weight in (2.75 +/- 0.25) kg, were divided randomly into model group and the sham operation group with fifteen rabbits in each group. The hibateral sternocleidomastoid muscles of rabbits in the model group were shortened by medical pipe to estabish the new animal model (the model was cervical dynamic dysequilibrium); and in the sham operative group, only exposed hibateral sternocleidomastoid muscles by operation. At the same time after two months, the histologic changes of the cervical muscles and intervertebral discs in all rabbits were observed, meanwhile, the myofibrillar amount and its cross section area were compared between two groups.

RESULTSAfter operation, the cervical muscles and intervertebral discs had significant change in model group, but no obvious change in sham operative group. The myofibrillar amount of frontal cervical muscles and back cervical muscles in model group was obviously lower than that of sham operative group (P < 0.05); likewise, the myofibrillar cross section area in model group was obviously lower than that of sham operative group (P < 0.05); the frontal cervical muscles was obviously change than the back cervical muscles.

CONCLUSIONThe cervical dynamic dysequilibrium caused by crispation of frontal cervical muscles can lead to pathologic degeneration of cervical muscles and intervertebral discs. The study may provide experimental proof for early cervical spondylopathy.

Animals ; Female ; Intervertebral Disc ; pathology ; Male ; Neck Muscles ; pathology ; physiology ; Rabbits ; Spasm ; pathology ; physiopathology

OBJECTIVETo study the level of serum vascular endothelial growth factor, matrix metalloproteinases-9 in the proliferative hemangioma before and after propranolol treatment.

METHODSThe serum VEGF, MMP-9 was detected with ELISA assay before treatment and after 4 weeks and 8 weeks of propranolol treatment. The relationship between the serum VEGF, MMP-9 and the prognosis was analyzed.

RESULTSThe serum VEGF (295.4 +/- 158.1) pg/ml was high before treatment, then decreased after 4 weeks and 8 weeks of treatment (255.7 +/- 130.4) pg/ml, (224.2 +/- 120.6) pg/ml. The serum VEGF was significantly lower after 8 weeks of treatment (P < 0.05). The serum MMP-9 was also decreased after treatment, showing a positive relationship with VEGF.

CONCLUSIONSPropranolol can treat the proliferative hemangioma through decreasing the serum VEGF and MMP-9.

Female ; Hemangioma ; blood ; drug therapy ; pathology ; Humans ; Infant ; Male ; Matrix Metalloproteinase 9 ; blood ; Propranolol ; therapeutic use ; Serum ; metabolism ; Vascular Endothelial Growth Factor A ; blood

OBJECTIVETo evaluate the prognostic value of surgical operation-related factors in patients with hepatocellular carcinoma (HCC).

METHODSThe clinical data of 234 patients after hepatic resection (214 men and 20 women) were retrospectively studied. Univariate and multivariate COX regression analyses were performed for surgical operation-related prognostic factors including age, gender, intraoperative blood loss, iatrogenic tumour rupture, transfusion, operation duration, hepatectomy extent, Pringle manoeuvre, with or without devarscularization, and complications (e.g. postoperative ascites, biliary leakage, incision infection, and pleural effusion). Kaplan-Meier and log-rank tests were used to compare survival rates. Kendall's tau bivariate analyses were used to examine the correlations of these surgical operation-related factors.

RESULTSUnivariate COX regression analysis revealed that iatrogenic blood loss (chi2 = 19.721, P < 0.001), transfusion (chi2 = 7.769, P = 0.005), tumour rupture (chi2 = 6.401, P = 0.011), operation duration (chi2 = 4.793, P = 0.029), and postoperative ascites (chi2 = 4.452, P = 0.035) were statistically significant predictors in patients with HCC after hepatic resection. Multivariate COX regression analysis revealed that pathological factors, such as blood loss (RR: 2.138, 95% CI: 1.556-2.939), tumour rupture (RR: 2.260, 95% CI: 1.182-4.321), and postoperative ascites (RR: 1.648, 95% CI: 1.088-2.469), independently influenced the HCC prognosis. Blood loss correlated with transfusion (Kendall's tau = 0.416, P < 0.001). There was no correlation between hepatectomy extent and blood loss (Kendall's tau = 0.057, P = 0.383), while transfusion closely correlated with the hepatectomy extent (Kendall's tau = 0.185, P = 0.004). The postoperative ascites closely correlated with Child classification (Kendall's tau = 0.151, P = 0.024).

CONCLUSIONSThe long-term survival of patients with HCC after hepatectomy may be improved by avoiding blood loss and iatrogenic tumour rupture. The indications of blood transfusion may not be strictly obeyed in some severe cases. Child class B and C cirrhotic patients may experience postoperative ascites and a worse prognosis, and therefore may be candidates for liver transplantation.

Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; mortality ; pathology ; surgery ; Child ; Female ; Hepatectomy ; adverse effects ; Humans ; Intraoperative Complications ; Liver Neoplasms ; mortality ; pathology ; surgery ; Male ; Middle Aged ; Postoperative Complications ; Retrospective Studies ; Survival Rate ; Treatment Outcome ; Young Adult