OBJECTIVETo analyze clinical features and genetic mutations in a Chinese family affected with autosomal dominant caveolinopathies.

METHODSClinical data of the proband and her family members were collected. Genomic DNA was extracted from peripheral blood samples with a standard procedure. Next generation sequencing was carried out for the proband, and direct sequencing was employed to detect potential mutation of the CAV gene.

RESULTSThe proband presented with slowly progressing distal muscle weakness and atrophy, especially distal upper limbs and muscular soreness during early childhood, with her CK level moderately elevated and EMG showing myogenic and neurogenic injuries. Her sisters presented mild symptoms with hand muscle atrophy and fasciculation after exercise. A heterozygous missense mutation c.80G>A (p.Arg27Gln), which was reported as being pathogenic, was identified in the CAV3 gene in the proband and her sisters.

CONCLUSIONA heterozygous c.80G>A (p.Arg27Gln) mutation in the CAV3 gene probably underlies the autosomal dominant caveolinopathies in this Chinese family.

Caveolin 3 ; genetics ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Middle Aged ; Muscular Dystrophies ; genetics ; Mutation

Objective: To investigate whether androgens can regulate the expression of eNOS in rat corpus cavernosum through AKT3, PIK3CA, CALM, and CAV1 and influence erectile function. METHODS: Thirty-six 8-week-old male SD rats were randomly divided into groups A (4-week control), B (6-week control), C (4-week castration), D (6-week castration), E (4-week castration + testosterone replacement), and F (6-week castration + testosterone replacement). Both the testis and epididymis were removed from the rats in groups C, D, E and F, and on the second day after surgery, the animals of groups E and F were subcutaneously injected with testosterone propionate at 3 mg per kg of the body weight qd alt while all the others with isodose oil instead. At 4 weeks (for groups A, C and E) and 6 weeks (for groups B, D and F) after treatment, we detected the maximum intracavernous pressure (ICPmax), the mean carotid arterial pressure (MAP) and their ratio (ICPmax/MAP), measured the level of serum testosterone (T), and determined the expressions of eNOS, P-eNOS, AKT3, PIK3CA, CALM and CAV1 in the corpus cavernosum by Western blot and immunohistochemistry. RESULTS: No statistically significant differences were observed in the body weight and MAP among different groups. The serum T level and ICPmax/MAP were remarkably lower in groups C and D than in the other four groups (P<0.01) as well as in groups E and F than in A and B (P<0.05) but exhibited no significant differences either between E and F or between A and B. Immunohistochemistry showed that eNOS and P-eNOS were mainly expressed in the vascular endothelial cell membrane and cavernous vascular lumen, while AKT3, PIK3CA, CALM and CAV1 chiefly in the vascular endothelial cell cytoplasm and membrane, with a few in the smooth muscle cells. Western blot analysis manifested that the expressions of eNOS, P-eNOS, AKT3, PIK3CA, CALM and CAV1 were markedly lower in groups C and D than in A, B, E and F (P<0.01) as well as in D than in C (P<0.05) but those in groups E and F did not showed any significant difference from those in A and B, nor E from F or A from B. CONCLUSIONS Androgens can improve erectile function by upregulating the expressions of AKT3, PIK3CA, CALM and CAV1 protein molecules and activating eNOS after its phosphorylation, though the exact molecular mechanisms are yet to be further studied.
Animals ; Blood Pressure ; Blotting, Western ; Caveolin 1 ; metabolism ; Class I Phosphatidylinositol 3-Kinases ; metabolism ; Erectile Dysfunction ; Hormone Replacement Therapy ; Male ; Monomeric Clathrin Assembly Proteins ; metabolism ; Myocytes, Smooth Muscle ; Nitric Oxide Synthase Type III ; metabolism ; Orchiectomy ; Penile Erection ; physiology ; Penis ; enzymology ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Testosterone Propionate ; administration & dosage

Porphyromonas gingivalis is one of the most important periodontal pathogens and has been to known to invade various types of cells, including endothelial cells. The present study investigated the mechanisms involved in the internalization of P. gingivalis in human umbilical vein endothelial cells (HUVEC). P. gingivalis internalization was reduced by clathrin and lipid raft inhibitors, as well as a siRNA knockdown of caveolin-1, a principal molecule of lipid raft-related caveolae. The internalization was also reduced by perturbation of actin rearrangement, while microtubule polymerization was not required. Furthermore, we found that Src kinases are critical for the internalization of P. gingivalis into HUVEC, while neither Rho family GTPases nor phosphatidylinositol 3-kinase are required. Taken together, this study indicated that P. gingivalis internalization into endothelial cells involves clathrin and lipid rafts and requires actin rearrangement associated with Src kinase activation.
Actins ; Caveolae ; Caveolin 1 ; Clathrin* ; Endothelial Cells* ; GTP Phosphohydrolases ; Human Umbilical Vein Endothelial Cells ; Humans ; Microtubules ; Phosphatidylinositol 3-Kinase ; Phosphotransferases ; Polymerization ; Polymers ; Porphyromonas gingivalis* ; RNA, Small Interfering ; src-Family Kinases

Exercise training can improve strength and lead to adaptations in the skeletal muscle and nervous systems. Skeletal muscles can develop into two types: fast and slow, depending on the expression pattern of myosin heavy chain (MHC) isoforms. Previous studies reported that exercise altered the distribution of muscle fiber types. It is not currently known what changes in the expression of caveolins and types of muscle fiber occur in response to the intensity of exercise. This study determined the changes in expression of caveolins and MHC type after forced exercise in muscular and non-muscular tissues in rats. A control (Con) group to which forced exercise was not applied and an exercise (Ex) group to which forced exercise was applied. Forced exercise, using a treadmill, was introduced at a speed of 25 m/min for 30 min, 3 times/day (07:00, 15:00, 23:00). Homogenized tissues were applied to extract of total RNA for further gene analysis. The expression of caveolin-3 and MHC2a in the gastrocnemius muscle of female rats significantly increased in the Ex group compared with the Con group (P<0.05). Furthermore, in the gastrocnemius muscle of male rats, the expression of MHC2x was significantly different between the two groups (P<0.05). There was an increased expression in caveolin-3 and a slightly decreased expression in TGFbeta-1 in muscular tissues implicating caveolin-3 influences the expression of MHC isoforms and TGFbeta-1 expression. Eventually, it implicates that caveolin-3 has positive regulatory function in muscle atrophy induced by neural dysfunction with spinal cord injury or stroke.
Animals ; Caveolin 3 ; Caveolins ; Female ; Humans ; Male ; Muscle, Skeletal ; Muscles ; Muscular Atrophy ; Myosin Heavy Chains ; Myosins ; Nervous System ; Protein Isoforms ; Rats ; RNA ; Spinal Cord Injuries ; Stroke

We investigated the effect of phenylephrine (PE)- and isoproterenol (ISO)-induced cardiac hypertrophy on subcellular localization and expression of caveolin-3 and STAT3 in H9c2 cardiomyoblast cells. Caveolin-3 localization to plasma membrane was attenuated and localization of caveolin-3 to caveolae in the plasma membrane was 24.3% reduced by the catecholamine-induced hypertrophy. STAT3 and phospho-STAT3 were up-regulated but verapamil and cyclosporin A synergistically decreased the STAT3 and phospho-STAT3 levels in PE- and ISO-induced hypertrophic cells. Both expression and activation of STAT3 were increased in the nucleus by the hypertrophy. Immunofluorescence analysis revealed that the catecholamine-induced hypertrophy promoted nuclear localization of pY705-STAT3. Of interest, phosphorylation of pS727-STAT3 in mitochondria was significantly reduced by catecholamine-induced hypertrophy. In addition, mitochondrial complexes II and III were greatly down-regulated in the hypertrophic cells. Our data suggest that the alterations in nuclear and mitochondrial activation of STAT3 and caveolae localization of caveolin-3 are related to the development of the catecholamine-induced cardiac hypertrophy.
Animals ; Catecholamines/*pharmacology ; Caveolae/*metabolism ; Caveolin 3/*metabolism ; Cell Line ; Hypertrophy/metabolism ; Mitochondria/*metabolism ; Myocardium/cytology/*pathology ; Myocytes, Cardiac/cytology/*drug effects/metabolism ; Rats ; STAT3 Transcription Factor/*metabolism

OBJECTIVETo study the relationship between expression of caveolin-1 (Cav-1) and pERK1/2 and prognosis in non-small cell lung cancer (NSCLC).

METHODSCav-1 and pERK1/2 protein expression was assessed by immunohistochemistry in samples obtained from 160 patients with NSCLC and 20 patients with normal lung tissue.

RESULTSNormal bronchial and alveolar epithelial cells were positive for Cav-1 (membranous and cytoplasmic staining patterns). The expression rate of Cav-1 in NSCLC was 65.6% (105/160), which was significantly lower than that in normal lung tissue (P = 0.002). The Cav-1-positive rates in well to moderately differentiated tumors and poorly differentiated tumors were 56.8% (46/81) and 75.7% (53/70), respectively (P = 0.015). The expression of Cav-1 was much higher in patients with lymph node metastasis (77.8%, compared with 55.7% in lymph node-negative group, P = 0.003). The expression was also higher in stage III to IV than in stage I to II disease (75.4%, compared with 58.2%, P = 0.024). The overall survival of patients with Cav-1-positive tumors (71.4%, 37.1% and 17.1% 1-, 3- and 5-year survival, respectively) was lower than those with Cav-1-negative tumors (89.1%, 69.1% and 43.6% 1-, 3- and 5-year survival, respectively, P = 0.000). On the other hand, normal bronchial and alveolar epithelial cells were negative for pERK1/2. The expression rate of pERK1/2 in NSCLC was 61.3%, which was significantly higher than that in normal lung tissues (P = 0.000). The pERK1/2-positive rates in well to moderately differentiated tumors and poorly differentiated tumors was 53.1% and 71.4%, respectively (P = 0.021). The expression of pERK1/2 was much higher in patients with lymph node metastasis (80.6%, compared with 45.5% in lymph node-negative group, P = 0.000). The expression of pERK1/2 was also higher in stage III to IV than in stage I to II disease (76.8%, compared with 49.5%, P = 0.426). The overall survival of patients with pERK1/2-positive tumors (74.5%, 42.9% and 19.4% 1-, 3- and 5-year survival, respectively) was lower than those with pERK1/2-negative tumors (82.3%, 56.5% and 37.1% 1-, 3- and 5-year survival, respectively, P = 0.002). Cav-1 and pERK1/2 expression showed negative correlation (P = 0.000).

CONCLUSIONSCav-1 expression is lower in NSCLC than in normal lung tissue, whereas pERK1/2 expression is higher in NSCLC. Positive expression of Cav-1 and overexpression of pERK1/2 correlates with tumorigenesis and tumor progression of NSCLC. Cav-1 and pERK1/2 may serve as potential markers for predicting prognosis in NSCLC.

Carcinoma, Non-Small-Cell Lung ; diagnosis ; metabolism ; Caveolin 1 ; genetics ; metabolism ; Cytoplasm ; Humans ; Immunohistochemistry ; methods ; Lung Neoplasms ; diagnosis ; metabolism ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; pathology ; Mitogen-Activated Protein Kinase 1 ; genetics ; metabolism ; Mitogen-Activated Protein Kinase 3 ; genetics ; metabolism ; Neoplasm Staging ; classification ; Prognosis

OBJECTIVETo investigate the role of caveolin-1 down-regulation in human hepatocyte proliferation in vitro.

METHODSThe expression vector psiRNA-CAV1 was constructed and transfected into Chang liver cells (CHL). The caveolin-1 down-regulated cell clones were selected by the antibiotic zeocin. The proliferation of the cell strain CAV7 was examined by MTT, in which untransfected CHL and HepG2 cells were set as controls. Expression of caveolin-1, Akt, Erk1/2, p-Akt and p-Erk1/2 in the transfected and control cells was detected by Western blot.

RESULTSAfter caveolin-1 expression was down-regulated by RNAi, CHL increased faster at first (24 h and 72 h, P<0.05; 96 h, P<0.01), but slower later. P-Akt and p-Erk1/2 expressions were down-regulated, indicating that the growth and proliferation related Akt and Erk1/2 pathways were inhibited after caveolin-1 down-regulation.

CONCLUSIONCaveolin-1 may play an important role in hepatocyte proliferation.

Caveolin 1 ; genetics ; metabolism ; Cell Line ; Cell Proliferation ; Down-Regulation ; Hepatocytes ; cytology ; Humans ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA Interference ; RNA, Small Interfering ; Signal Transduction ; Transfection

It is well known that exercise can have beneficial effects on insulin resistance by activation of glucose transporter. Following up our previous report that caveolin-1 plays an important role in glucose uptake in L6 skeletal muscle cells, we examined whether exercise alters the expression of caveolin-1, and whether exercise-caused changes are muscle fiber and exercise type specific. Fifity week-old Sprague Dawley (SD) rats were trained to climb a ladder and treadmill for 8 weeks and their soleus muscles (SOL) and extensor digitorum longus muscles (EDL) were removed after the last bout of exercise and compared with those from non-exercised animals. We found that the expression of insulin related proteins and caveolins did not change in SOL muscles after exercise. However, in EDL muscles, the expression of insulin receptor beta (IRbeta) and glucose transporter-4 (GLUT-4) as well as phosphorylation of AKT and AMPK increased with resistance exercise but not with aerobic exercise. Also, caveolin-1 and caveolin-3 increased along with insulin related proteins only in EDL muscles by resistance exercise. These results suggest that upregulation of caveolin-1 in the skeletal muscle is fiber specific and exercise type specific, implicating the requirement of the specific mode of exercise to improve insulin sensitivity.
AMP-Activated Protein Kinases ; Animals ; Caveolin 1/*biosynthesis ; Caveolin 3/metabolism ; Female ; Glucose Transporter Type 4/biosynthesis ; Insulin/*physiology ; Multienzyme Complexes/metabolism ; Muscle Fibers, Skeletal/*metabolism ; Muscle, Skeletal/metabolism/*physiology ; Phosphorylation ; *Physical Conditioning, Animal ; Protein-Serine-Threonine Kinases/metabolism ; Proto-Oncogene Proteins c-akt/metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Insulin/biosynthesis ; Up-Regulation

OBJECTIVETo investigate the effects of caveolin-1 on the biologic behavior of laryngeal squamous cell carcinoma HEp2 cell line in vitro.

METHODSEukaryotic expression vector of human caveolin-1 gene was constructed and transfected into HEp2 cells by Lipofectamine. The clones stably overexpressing caveolin-1 were identified by real-time PCR and Western blotting. Cell proliferation viability was tested by MTT assay. Anchorage-independent growth was determined by assaying colony formation in soft agar. Flow cytometry was used to assess the cell cycle and apoptosis. The relative phosphorylation level of EGFR and ERK1/2 were detected by Western blotting. Localization of caveolin-1 and EGFR were studied by laser confocal laser scanning microscopy.

RESULTSThe expression vector of caveolin-1 was constructed and three clones stably overexpressing caveolin-1 were obtained. Comparing with the parental HEp2 cells, the transfected cells exhibited a slower growth rate and formed fewer colonies in soft agar. The results of FACS analysis revealed that overexpression of caveolin-1 resulted in the cell cycle arrest at G0/G1 phase and increased the apoptotic cell fraction. EGFR was found to colocalize with caveolin-1 in transfected cells by confocal laser scanning microscopy and Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of EGFR and Erkl/2.

CONCLUSIONOverexpression of caveolin-1 suppresses the growth of HEp2 cells and induces apoptosis and inhibition of EGFR-MAPK signaling pathway may be involved in its mechanism.

Apoptosis ; Blotting, Western ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Caveolin 1 ; genetics ; metabolism ; physiology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; Flow Cytometry ; Genetic Vectors ; chemistry ; genetics ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; Lipids ; chemistry ; Microscopy, Confocal ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Polymerase Chain Reaction ; methods ; Receptor, Epidermal Growth Factor ; metabolism ; Signal Transduction ; physiology ; Transfection ; methods

The purpose of this study was to demonstrate the cellular localization of cyclooxygenase-2 (COX-2) and caveolin-3 (Cav-3) in primarily cultured rat chondrocytes. In normal rat chondrocytes, we observed relatively high levels of Cav-3 and a very low level of COX-2 mRNA and protein. Upon treating the chondrocytes with 5 microM of CdCl2 (Cd) for 6 hr, the expressions of COX-2 mRNA and protein were increased with the decreased Cav-3 mRNA and protein expressions. The detergent insoluble caveolae-rich membranous fractions that were isolated from the rat chondrocytes and treated with Cd contained the both proteins of both COX-2 and Cav-3 in a same fraction. The immuno-precipitation experiments showed complex formation between the COX-2 and Cav-3 in the rat chondrocytes. Purified COX-2 with glutathione S-transferase-fused COX-2 also showed complex formation with Cav-3. Confocal and electron microscopy also demonstrated the co-localization of COX-2 and Cav-3 in the plasma membrane. The results from our current study show that COX-2 and Cav-3 are co-localized in the caveolae of the plasma membrane, and they form a protein-protein complex. The co-localization of COX-2 with Cav-3 in the caveolae suggests that the caveolins might play an important role for regulating the function of COX-2.
Animals ; Animals, Newborn ; Blotting, Western ; Cadmium Chloride/pharmacology ; Caveolae/drug effects/metabolism/ultrastructure ; Caveolin 3/*genetics/metabolism ; Cell Membrane/drug effects/metabolism ; Cells, Cultured ; Chondrocytes/cytology/drug effects/*metabolism ; Cyclooxygenase 2/*genetics/metabolism ; Gene Expression ; Immunoprecipitation ; Microscopy, Confocal ; Microscopy, Electron ; RNA, Messenger/genetics/metabolism ; Rats ; Reverse Transcriptase Polymerase Chain Reaction