OBJECTIVETo investigate the effect of anti-miR-145 on human airway smooth muscle cell (HASMC) proliferation and osteopontin systhesis in vitro and explore the mechanisms.

METHODSHASMCs were treated with 10-100 nmol/L anti-miR-145, and the cell proliferation and apoptosis were investigated using a CCK-8 assay and flow cytometry, respectively. The changes in osteopontin synthesis after the treatment was quantified with Western blotting.

RESULTSTreatment with 10 and 50 nmol/L anti-miR-145 significantly promoted the proliferation and osteopontin synthesis in HASMCs (P<0.05 or <0.01), and 50 nmol/L anti-miR-145 obviously inhibited the cell apoptosis (P<0.01).

CONCLUSIONAnti-miR-145 promotes HASMC proliferation and osteopontin synthesis and inhibits HASMC apoptosis in vitro, indicating the important role of anti-miR-145 in the pathogenesis of airway remodeling.

Airway Remodeling ; Apoptosis ; Cell Proliferation ; Cells, Cultured ; Humans ; MicroRNAs ; antagonists & inhibitors ; Myocytes, Smooth Muscle ; drug effects ; Osteopontin ; biosynthesis ; Respiratory System ; cytology

OBJECTIVETo determine the expression of osteopontin (OPN) and survivin in prostate cancer tissue, and study their correlation and roles in tumor invasion and metastasis.

METHODSThe expressions of OPN and survivin in prostate cancer tissue, prostate hyperplasia tissue and normal prostate tissue were determined by RT-PCR and immunohistochemistry.

RESULTSThe positive expression rates of OPN mRNA and protein in prostate cancer tissue [76.1% (35/46) and 69.6% (32/46)] were significantly correlated to survivin expression [67.4% (31/46) and 67.4% (31/46)] (P<0.05). The expressions of OPN and survivin were related to the tumor grade and clinical stages (P<0.05). OPN and survivin were not found in prostate hyperplasia and normal prostate tissues.

CONCLUSIONOPN and survivin may play important roles in the progression of prostate cancer and can be potential markers for invasion and metastasis of prostate cancer. OPN and survivin might play synergetic roles in prostate carcinogenesis.

Adult ; Aged ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Osteopontin ; biosynthesis ; genetics ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism

OBJECTIVETo investigate osteopontin (OPN) mRNA expression in oral squamous cell carcinoma (OSCC) and normal oral mucosa tissues.

METHODSDifferential OPN gene expression were detected in 30 cancerous tissues and their paired normal tissues using real-time reverse transcription-PCR (real-time RT-PCR), and the data were analyzed statistically.

RESULTSReal-time RT-PCR results demonstrated that the relative expression level of OPN mRNA in the cancerous tissues were significantly higher than that in paired normal samples (4.17-/+0.51 vs 0.97-/+0.12, P<0.001), showing a 4.3-fold up-regulation. In the 30 OSCC specimens, OPN mRNA expression in the OSCC of histological grades I showed a 3.1-fold down-regulation, significantly lower than the expression in grade II/III tumors (2.16-/+0.17 vs 6.80-/+0.72, P<0.05); its expression was significantly lower in early stage than in advanced stage OSCCs (2.34-/+0.17 vs 4.73-/+0.35, P<0.05). In cases of cervical lymph node metastasis, the expression was significantly higher than that in cases without lymphatic metastasis (6.38-/+0.56 vs 2.89-/+0.32, P<0.05).

CONCLUSIONOPN mRNA overexpression may play an important role in OSCC carcinogenesis and can be a potential target for OSCC therapy.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; genetics ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Mouth Mucosa ; metabolism ; pathology ; Mouth Neoplasms ; genetics ; pathology ; Osteopontin ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

The expression of osteopontin (OPN) in boar testis was studied. Western blot analysis detected 66- and 32-kDa OPN immunopositive bands in the testes of adult boars. In postnatal piglets, the 66-kDa OPN band was detected in the testes, but not the 32-kDa band. In the newborn testis, OPN immunostaining was seen in gonocytes and in some supporting cells in the seminiferous tubules, as well as in interstitial Leydig cells. In the adult boar testis, OPN immunoreactivity was detected in seminiferous tubules with varying intensities. Intense OPN immunostaining was seen in the residual bodies and acrosomes in the spermatids while, occasionally, OPN immunostaining was seen in spermatogonia and various stage of spermatocytes but in few Sertoli cells in the seminiferous tubules. In addition, Leydig cells in adult boars were weakly immunostained with OPN. These findings suggest that OPN is detected in the majority of germ cells and is involved in spermatogenesis in boar testis.
Animals ; Animals, Newborn ; Blotting, Western/veterinary ; Immunohistochemistry/veterinary ; Leydig Cells/metabolism ; Male ; Osteopontin/*biosynthesis ; Sertoli Cells/metabolism ; Spermatogenesis/physiology ; Spermatozoa/metabolism ; Swine/*metabolism ; Testis/cytology/*metabolism

Adenocarcinoma ; metabolism ; pathology ; Adenocarcinoma, Mucinous ; metabolism ; pathology ; Adenocarcinoma, Papillary ; metabolism ; pathology ; Carcinoma, Signet Ring Cell ; metabolism ; pathology ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Organic Cation Transport Proteins ; biosynthesis ; genetics ; Organic Cation Transporter 2 ; Osteopontin ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Sialoglycoproteins ; biosynthesis ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Survival Rate

OBJECTIVETo investigate the expressions of RhoC and osteopontin (OPN) protein in esophageal squamous carcinoma (ESC) and their association with the biological behavior of ESC.

METHODSThe expressions of RhoC and OPN protein were detected in 80 ESC cases by immunohistochemistry.

RESULTSThe positive expression rate of RhoC was 66.25% in these ESC cases. The rate was significantly higher in cases with lymph node metastasis than in those without (r(s)=-2.115, P<0.05), but RhoC expression was not associated with the tumor diameter, differentiation or TNM grade (P>0.05). The RhoC-positive patients had significantly shorter survival time than the negative patients (P<0.001). All the 80 ESC patients were positive for OPN expression, and OPN expression levels were correlated with the differentiation (chi(2)=10.766, P<0.05) and lymph node metastasis of the tumor (r(s)=-2.289, P<0.05), but not with the tumor diameter or TNM grade (P>0.05). Higher expression level of OPN was closely related to shorter survival time of the patients (P<0.05). A positive correlation was found between RhoC protein and OPN expressions (r(s)= 0.408, P<0.001) in these cases.

CONCLUSIONThe expressions of RhoC and OPN protein are closely related to lymph node metastasis of ESC and the patients survival time, and therefore may serve the purpose of prognostic evaluation of ESC.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; biosynthesis ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Osteopontin ; biosynthesis ; Prognosis ; Survival Analysis ; rho GTP-Binding Proteins ; biosynthesis ; rhoC GTP-Binding Protein

OBJECTIVETo observe the effects of perindopril on left ventricular remodeling and myocardial osteopontin expression in rats with myocardial infarction (MI).

METHODSMale adult SD rats were randomly divided into sham-operation group, MI-saline group and MI-perindopril group, and in the latter two groups, ligation of the left anterior descending artery was performed to establish rat models of myocardial infarction and perindopril (2 mg/kg daily) or saline was administered since the next day of MI. Four weeks later, the left ventricular diameter (LVEDD and LVESD) and left ventricular ejection fraction were estimated with echocardiography, and the left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP) and -/+dp/dt(max) was obtained via hemodynamic measurement, with also evaluation of the cardiac myocyte diameter and interstitial fibrosis infiltration with histological methods. Western blotting was performed to detect the expression level of myocardium osteopontin protein.

RESULTSCompared with the sham-operation group, all MI rats developed significant systolic and diastolic dysfunction, as indicated by decreased LVEF, LVSP and -/+dp/dt(max), as well as increased LVEDP. MI rats showed significantly dilated left ventricles and higher ventricular weight/body weight ratio, significantly increased cardiomyocyte diameter and marked interstitial fibrosis in the non-infarction area. Perindopril treatment partially prevented cardiac dysfunction and left ventricular remodeling as indicated by the above indices. No osteopontin was detected in the myocardium of sham-operation rats, and in MI rats, high level of osteopontin expression, as detected in MI-saline group, was significantly inhibited by perindopril treatment.

CONCLUSIONPerindopril treatment can significantly inhibit left ventricular remodeling and myocardium osteopontin expression in rats with MI.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Blotting, Western ; Heart ; drug effects ; physiopathology ; Male ; Myocardial Infarction ; physiopathology ; Myocardium ; metabolism ; Osteopontin ; biosynthesis ; Perindopril ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Ventricular Remodeling ; drug effects

OBJECTIVETo study the effect of osteopontin (OPN) expression down-regulated by RNA interference (RNAi) on the invasiveness of hepatocelluar carcinoma cell line HCC-LM3.

METHODSHCC-LM3 cells were transfected with the chemically synthesized small interfering RNA (siRNA) formulated by lipofectamine 2000. Wild type HCC-LM3 and HCC-LM3 cells transfected with non-specific siRNA served as controls. Real-time PCR and Western blotting were used to quantify the mRNA and OPN protein levels. The malignant phenotypes of transfected HCC-LM3 cells including cellular growth rate, colony formation and Matrigel invasion activities were analyzed.

RESULTSSequence-specific siRNAs targeting OPN suppressed OPN RNA expression by 79% and also decreased OPN protein level by 81% in HCC-LM3 cells. The number of formed colonies and migrating numbers in vitro were decreased in HCC-LM3 cells transfected using sequence-specific siRNAs targeting OPN relative to controls (P < 0.05).

CONCLUSIONThis study demonstrated that specific siRNA is able to reduce OPN at both the mRNA and protein levels and significantly diminishes the invasiveness of hepatocellular carcinoma cells.

Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Down-Regulation ; Humans ; Liver Neoplasms ; genetics ; pathology ; Neoplasm Invasiveness ; Osteopontin ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVE: To investigate the expression and significance of matrix metalloproteinase-3 (MMP-3), osteopontin (OPN) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in human lens epithelial cells (LECs) of cataract. METHODS: The MMP-3, OPN, and TIMP-1 expressions in LECs of anterior subcapsular cataract (31 cases), nuclear cataract (28 cases) and normal lens (12 cases) were detected by immunohistochemistry, respectively. RESULTS: The MMP-3 expression in anterior subcapsular cataract was significantly higher than that in nuclear cataract and normal lens (chi2=31.49, 34.479; P=0.000); but there was no statistic significance between nuclear cataract and normal lens (chi2=2.449, P=0.118). The OPN expression in anterior subcapsular cataract was also significantly higher than that in nuclear cataract and normal lens (chi2=29.450, 15.889; P=0.000). There was no significant difference in the TIMP-1 expression among the 3 groups (P>0.05). Positive correlation was found between MMP-3 and OPN in LECs of anterior subcapsular cataract (r=0.381, P=0.035). But no significant correlation was found among MMP-3, OPN, and TIMP-1 (r=0.121, -0.289; P=0.516, 0.114). CONCLUSION Increased expression of MMP-3 and OPN and the expression imbalance between MMP-3 and TIMP-1 may play a critical role in the pathogenesis of anterior subcapsular cataract.
Adult ; Aged ; Aged, 80 and over ; Cataract ; classification ; etiology ; metabolism ; Epithelial Cells ; metabolism ; Female ; Humans ; Lens, Crystalline ; metabolism ; Male ; Matrix Metalloproteinase 3 ; biosynthesis ; Middle Aged ; Osteopontin ; biosynthesis ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis

Conditions of disuse such as bed rest, space flight, and immobilization result in decreased mechanical loading of bone, which is associated with reduced bone mineral density and increased fracture risk. Mechanisms involved in this process are not well understood except the suppression of osteoblast function. To investigate the effect of simulated weightlessness on mRNA level of extracellular matrix proteins, osteoblasts were rotated in horizontal plane as a model of simulated microgravity. Primordial osteoblasts of rats were grown for 2 d and then rotated for 24, 48 and 72 h, respectively. After isolating total RNA in cells, reverse transcription polymerase chain reaction (PT-PCR) was made to examine the mRNA level of osteopontin (OPN) and osteonectin (ON). Meanwhile, the levels of alkaline phosphatase (ALP) and osteocalcin (BGP) in the cultured medium were measured to evaluate the calcific function of cell. The expression of OPN and ON mRNA fell significantly after rotating for 24, 48 and 72 h, respectively. The contents of BGP descended significantly, meanwhile, the activity of ALP also showed a degressive tendency. Horizontal rotation decreased the expression of ON and OPN as well as diminished the secretion of BGP and ALP, which affected the calcific function of osteoblast. The results obtained suggest that depression of extracellular matrix proteins expression plays a key role in bone loss during weightlessness.
Animals ; Animals, Newborn ; Cells, Cultured ; Extracellular Matrix Proteins ; biosynthesis ; genetics ; Osteoblasts ; cytology ; metabolism ; Osteonectin ; biosynthesis ; genetics ; Osteopontin ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Rotation ; Sialoglycoproteins ; biosynthesis ; genetics ; Skull ; cytology ; Weightlessness Simulation