ABSTRACT
BACKGROUND: Extracellular sulfatases (Sulfs), sulfatase 1 (Sulf1) and sulfatase 2 (Sulf2), play a pivotal role in cell signaling by remodeling the 6-O-sulfation of heparan sulfate proteoglycans on the cell surface. The present study examined the effects of Sulfs on angiotensin II (Ang II)-induced hypertensive mediator expression and vascular smooth muscle cells (VSMCs) proliferation in spontaneously hypertensive rats (SHR). METHODS: Ang II receptors, 12-lipoxygenase (12-LO), and endothelin-1 (ET-1) messenger RNA (mRNA) expressions in SHR VSMCs were analyzed by real-time polymerase chain reaction and Western blotting. VSMCs proliferation was determined by [³ H]-thymidine incorporation. RESULTS: Basal Sulfs mRNAs expression and enzyme activity were elevated in SHR VSMCs. However, Sulfs had no effect on the basal or Ang II-induced 12-LO and ET-1 mRNA expression in SHR VSMCs. The inhibition of Ang II-induced 12-LO and ET-1 expression by blockade of the Ang II type 2 receptor (AT₂ R) pathway was not observed in Sulf1 siRNA-transfected SHR VSMCs. However, Sulf2 did not affect the action of AT₂ R inhibitor on Ang II-induced 12-LO and ET-1 expression in SHR VSMCs. The down-regulation of Sulf1 induced a reduction of AT₂ R mRNA expression in SHR VSMCs. In addition, the inhibition of Ang II-induced VSMCs proliferation by blockade of the AT₂ R pathway was mediated by Sulf1 in SHR VSMCs. CONCLUSION: These findings suggest that extracellular sulfatase Sulf1 plays a modulatory role in the AT₂ R pathway that leads to an Ang II-induced hypertensive effects in SHR VSMCs.
Subject(s)
Angiotensin II , Angiotensins , Arachidonate 12-Lipoxygenase , Blotting, Western , Down-Regulation , Endothelin-1 , Heparan Sulfate Proteoglycans , Hypertension , Muscle, Smooth, Vascular , Rats, Inbred SHR , Real-Time Polymerase Chain Reaction , Receptor, Angiotensin, Type 2 , RNA, Messenger , SulfatasesABSTRACT
BACKGROUND: Extracellular sulfatases (Sulfs), sulfatase 1 (Sulf1) and sulfatase 2 (Sulf2), play a pivotal role in cell signaling by remodeling the 6-O-sulfation of heparan sulfate proteoglycans on the cell surface. The present study examined the effects of Sulfs on angiotensin II (Ang II)-induced hypertensive mediator expression and vascular smooth muscle cells (VSMCs) proliferation in spontaneously hypertensive rats (SHR).METHODS: Ang II receptors, 12-lipoxygenase (12-LO), and endothelin-1 (ET-1) messenger RNA (mRNA) expressions in SHR VSMCs were analyzed by real-time polymerase chain reaction and Western blotting. VSMCs proliferation was determined by [³ H]-thymidine incorporation.RESULTS: Basal Sulfs mRNAs expression and enzyme activity were elevated in SHR VSMCs. However, Sulfs had no effect on the basal or Ang II-induced 12-LO and ET-1 mRNA expression in SHR VSMCs. The inhibition of Ang II-induced 12-LO and ET-1 expression by blockade of the Ang II type 2 receptor (AT₂ R) pathway was not observed in Sulf1 siRNA-transfected SHR VSMCs. However, Sulf2 did not affect the action of AT₂ R inhibitor on Ang II-induced 12-LO and ET-1 expression in SHR VSMCs. The down-regulation of Sulf1 induced a reduction of AT₂ R mRNA expression in SHR VSMCs. In addition, the inhibition of Ang II-induced VSMCs proliferation by blockade of the AT₂ R pathway was mediated by Sulf1 in SHR VSMCs.CONCLUSION: These findings suggest that extracellular sulfatase Sulf1 plays a modulatory role in the AT₂ R pathway that leads to an Ang II-induced hypertensive effects in SHR VSMCs.
Subject(s)
Angiotensin II , Angiotensins , Arachidonate 12-Lipoxygenase , Blotting, Western , Down-Regulation , Endothelin-1 , Heparan Sulfate Proteoglycans , Hypertension , Muscle, Smooth, Vascular , Rats, Inbred SHR , Real-Time Polymerase Chain Reaction , Receptor, Angiotensin, Type 2 , RNA, Messenger , SulfatasesABSTRACT
Objective:To investigate the mRNA and protein expressions of steroid sulfatase (STS) in breast cancer tissues and normal breast tissues, and analyze its relationship with clinicopathologic characteristics. Methods:The mRNA and protein expressions of STS, in 40 cases of breast cancer tissues and corresponding paracancerous normal breast tissues, were examined by reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemistry. The correlation of STS expression level with clinicopathologic characteristics was analyzed. Results:STS protein was mainly expressed in the cytoplasm of breast carcinoma cells and epithelial cells in normal breast glands, but not in the stroma. It could be detected in the nucleus of carcinoma cells in 3 cases of breast cancer tissues, which was pathologically classified as invasive ductal carcinoma, invasive lobular carcinoma, and invasive micropapillary carcinoma. STS was not observed in interstitial tissues of breast glands. STS protein expression had positive correlation with its mRNA expressing level. The positivity of STS was 70.0% in breast cancer tissues, significantly higher than that of normal breast tissues (42.5%). The difference was significant (P =0.013). Stratified analysis showed that the positive rates of STS protein were significantly higher in premenopausal patients, the patients with lymph node metastasis, and those with advanced breast carcinoma than those in the matched normal breast tissues (P<0.05). Conclusion:Breast cancer tissues highly expressed STS protein to stimulate local estrogen production, thereby enhancing the progression and migration of breast cancer cells. In addition, as the tumor growth, locally biosynthesized estrogens may play more and more important roles.
ABSTRACT
Objective To explore the expression of estrogen sulfotransferase(EST) and steroid sulfatase (STS) in formalin-fixed paraffin-embedded endometrioid adenocarcinoma samples. Methods The RNA of EST and STS in 30 formalin-fixed paraffin-embedded endometrium samples were extracted using Roche products. Results The RNA expression of EST and STS were 0.25±0.03 and 0.08±0.02 respectively and the STS/EST was 0.11±0.08 in normal endometrium. While in endometrioid adenocarcinoma the RNA expression of EST and STS were 0.06±0.02 and 0.24±0.92 respectively and the STS/EST was 4.40±0.64. There were significant differences between these two groups. Conclusions (1) Formalin-fixed paraffin-embedded tissues could be used to study the endometrioid adenocarcinoma. (2) EST is decreased, STS and STS/EST are increased in human endometrioid adenocarcinoma. STS/EST may be related with the prognosis of the endometrioid adenocarcinoma.
ABSTRACT
Sulfoconjugates occur ubiquitously as sulfopolysaccharides, sulfolipids and sulfoproteins. A variety of sulfotransferases catalyze the sulfation process with 3'- phosphoadenosine 5'-phosphosulfate as the sulfate donor. Sulfatases that catalyze the desulfation of different sulfoconjugates are known to be deficient in a number of genetic storage disorders.