2.Effects of rotation on osteonectin and osteopontin mRNA level of cultured osteoblasts in rats.
Yu-Min WAN ; Yong-Jie MA ; Xiao-You ZHANG ; Bin ZENG ; Hong-Hui WANG ; Ying-Hui LI
Acta Physiologica Sinica 2005;57(3):384-388
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
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Animals, Newborn
;
Cells, Cultured
;
Extracellular Matrix Proteins
;
biosynthesis
;
genetics
;
Osteoblasts
;
cytology
;
metabolism
;
Osteonectin
;
biosynthesis
;
genetics
;
Osteopontin
;
RNA, Messenger
;
biosynthesis
;
genetics
;
Rats
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Rats, Wistar
;
Rotation
;
Sialoglycoproteins
;
biosynthesis
;
genetics
;
Skull
;
cytology
;
Weightlessness Simulation
3.Expressions of RhoC and osteopontin in esophageal squamous carcinoma and association with the patients' prognosis.
Hui-zhong ZHANG ; Jin-geng LIU ; Yi-ping WEI ; Cheng WU ; Yong-ke CAO ; Mei WANG
Journal of Southern Medical University 2006;26(11):1612-1615
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
4.Effects of matrix metalloproteinase-3, osteopontin, and tissue inhibitor of metalloproteinase-1 in the formation of cataract.
Qian TAN ; Jian JIANG ; Dong-qiang LUO
Journal of Central South University(Medical Sciences) 2006;31(5):737-741
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
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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
5.Immunohistochemical study of osteopontin in boar testis.
Journal of Veterinary Science 2007;8(2):107-110
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
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Animals, Newborn
;
Blotting, Western/veterinary
;
Immunohistochemistry/veterinary
;
Leydig Cells/metabolism
;
Male
;
Osteopontin/*biosynthesis
;
Sertoli Cells/metabolism
;
Spermatogenesis/physiology
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Spermatozoa/metabolism
;
Swine/*metabolism
;
Testis/cytology/*metabolism
6.Effect of astragalus-angelica mixture on osteopontin expression in rats with chronic nephrosclerosis.
Ya-ni ZHAO ; Jing-zi LI ; Ling YU
Chinese Journal of Integrated Traditional and Western Medicine 2002;22(8):613-617
OBJECTIVETo observe the effect of Astragalus-Angelica Mixture (AAM) on osteopontin (OPN) expression in rats with chronic nephrosclerosis.
METHODSChronic nephrosclerosis model rats induced by repeated intraperitoneal injection of puromycin were randomly divided into the model group, AAM group and Irbesartan (an antagonist of angiotensin) group. The experimental course lasted 12 weeks. Blood and urine samples were examined by biochemical method. Kidney tissue was taken for pathological stain and immunohistochemical method and was applied to examine OPN expression, mononuclear macrophage, laminin in extracellular matrix and decorin expressions.
RESULTSAAM showed the effects of decreasing urinary protein and improving renal function similar to that of Irbesartan. It also could alleviate the pathological damage of kidney tissue, especially in decreasing renal tubular mesenchymal damage index. The accumulation of decorin and laminin in the mesenchymal extracellular matrix significantly decreased. Renal tubular OPN expression and mesenchymal infiltration of mononuclear macrophage decreased significantly and in a positive correlated manner (r = 0.885, P < 0.01).
CONCLUSIONAAM has similar renal protective action to that of Irbesartan, this action may be related to the inhibition of up-regulated OPN expression.
Animals ; Astragalus Plant ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Male ; Nephrosclerosis ; chemically induced ; metabolism ; Osteopontin ; Phytotherapy ; Puromycin ; Rats ; Rats, Sprague-Dawley ; Sialoglycoproteins ; biosynthesis
7.Expression of osteopontin and survivin in prostate cancer and the clinical significance.
Hong-bo YU ; Xiao-bing HAN ; Yi-qian LIANG ; Jian-gang LIU ; Hao WANG
Journal of Southern Medical University 2010;30(5):1141-1143
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
8.Anti-miR-145 promotes human airway smooth muscle cell proliferation and osteopontin synthesis in vitro.
Peifen CHEN ; Zhihui QIU ; Guohua HUANG ; Xiangmei ZHANG ; Wujian PENG ; Hui CENG ; Wenyan LAI
Journal of Southern Medical University 2015;35(7):1073-1075
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
9.Expression of osteopontin mRNA and its clinical significance in gastric cancer.
Xian-jun SUN ; Wen-shu ZUO ; Heng MA ; Wen-hong HOU ; Shu-Ping CAI ; Xi-hong JIANG
Chinese Journal of Oncology 2005;27(5):292-295
OBJECTIVETo investigate the expression of osteopontin mRNA and its correlation with clinicopathologic features of gastric cancer and elucidate its role in tumor invasion and distant metastasis.
METHODSThe expression of OPN mRNA was detected by semi-quantitive RT-PCR. The relationship between the relative content of OPN mRNA and clinicopathologic features of gastric cancer was analyzed.
RESULTSIn 66 cancer tissue samples, a 330 bp band was detected in 50 cases, the positive rate of OPN mRNA expression was 75.8% (50/66). The expression in all 20 cases of normal gastric mucosa was negative. The expression was associated with the depth of tumor invasion, diameter, lymph node metastasis and but had no correlation with differentiation grades. The 66 patients were followed up for 10 approximately 27 months (mean 16 months). The OPN mRNA expression positive group (50 cases) had recurrence in 15 patients and the negative group (16 cases) had only 1 case with recurrence (P = 0.05); 10 patients died in OPN mRNA expression positive group but no patient died in OPN staining negative group (P = 0.05).
CONCLUSIONOPN mRNA is over-expressed in gastric cancer. It reflects the progression of disease and association with poor prognosis of gastric cancer. OPN may play an important role in the process of distant metastasis in gastric cancer.
Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Osteopontin ; biosynthesis ; genetics ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; metabolism ; pathology
10.Expression of osteopontin mRNA in oral squamous cell carcinoma and normal oral mucosa.
Xiao-zhi LU ; Xue-min YIN ; Yuan-yong FENG ; Li-qiong DUAN ; Wan-tao CHEN ; Chen-ping ZHANG
Journal of Southern Medical University 2008;28(7):1165-1167
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