Clinical Significance of SFRP1 Gene Methylation in Patients with Childhood Acute Lymphoblastic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical significance of SFRP1 gene and its methylation in childhood acute lymphoblastic leukemia (ALL) .@*METHODS@#Methylation-specific PCR (MSP) was used to detect the methylation status of SFRP1 gene in bone marrow mononuclear cells of 43 children with newly diagnosed ALL before chemotherapy (primary group) and when the bone marrow reached complete remission d 46 after induction of remission chemotherapy (remission group), the expression of SFRP1 mRNA was detected by quantitative real-time polymerase chain reaction (qRT-PCR), the expression of SFRP1 protein was detected by Western blot, and clinical data of children were collected, the clinical significance of SFRP1 gene methylation in children with ALL was analyze.@*RESULTS@#The positive rate of SFRP1 gene promoter methylation in the primary group (44.19%) was significantly higher than that in the remission group (11.63%) (χ2=11.328, P<0.05). The relative expression levels of SFRP1 mRNA and protein in bone marrow mononuclear cells of children in the primary group were significantly lower than those in the remission group (P<0.05). Promoter methylation of SFRP1 gene was associated with risk level (χ2=15.613, P=0.000) and survival of children (χ2=6.561, P=0.010) in the primary group, children with SFRP1 hypermethylation had significantly increased risk and shortened event-free survival time, but no significant difference in other clinical data.@*CONCLUSION@#Hypermethylation of SFRP1 gene promoter may be involved in the development of childhood ALL, and its hypermethylation may be associated with poor prognosis.

SFRP1, Posible biomarcador en la progresión o regresión de lesiones de cérvix asociado al Virus del Papiloma Humano / SFRP1, Possible Biomarker in the Progression or Regression of Cervical Pre-neoplastic Lesions Associated with Human Papilloma Virus

Resumen Objetivo: Comparar la expresión de mRNA y proteínas de SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 y HAND1 en pacientes con lesión intra-epitelial cervical de bajo y alto grado, con posterior progresión o regresión. Material y Método: Se realizó análisis de expresión de genes mediante RT-PCR y análisis de expresión de proteínas por inmunohistoquímica. El análisis estadís tico fue realizado con las pruebas: Wilcoxon, coeficiente de correlación de Spearman e índice de concordancia. Las muestras fueron pareadas en momento 1 y momento 2. Resultados: SFRP1 mostró tendencia de mayor expresión de mRNA en lesión intra-epitelial de bajo grado (momento 2) Vs. alto grado (momento 1). La expresión de proteínas por inmunohistoquímica de SFRP1 en casos de progresión (83,3 %) mostró disminución en su graduación (p = 0,0313*); los demás genes en estudio no tuvieron cambios estadísticamente significativos. Discusión: SFRP1 mostró comportamiento ajustado a resultados de estudios previos donde se encontró hipermetilado en lesiones intra-epiteliales de alto grado; su subexpresión por hipermetilación se reportó en otros canceres, proceso que colabora con su silenciamiento y transición epitelial-mesenquimatosa del cáncer de cuello uterino. Conclusiones. SFRP1 es potencial biomarcador en lesiones preneoplásicas del cuello uterino asociadas al virus de papiloma humano.

Abstract Objective. The aim of this work was to compare the expression of mRNA and proteins of SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 and HAND1 in patients with low and high grade cervical intraepithelial lesion, with subsequent progression or regression. Material and Methods: Gene expression analysis was conducted through RT-PCR and protein expression analysis was performed by immunohistochemistry. The statistics analysis were Wilcoxon test, Spearman's correlation coefficient and concordance index. The samples were paired during moment 1 (initial patient diag nosis) and moment 2 (follow-up histological diagnosis). Results: SFRP1 showed a trend of higher mRNA expression in low-grade intra-epithelial lesions (moment 2) Vs. high-grade (moment 1). The expression of proteins by immunohistochemistry of SFRP1 in progression cases (83.3%) showed a decrease in its graduation (p = 0.0313*); the other genes under study had no statistically significant. Discussion: SFRP1 showed a biological behavior adjusted to the results of previous studies where hypermethylation was found in high-grade intra-epithelial lesions; its subexpression by hypermethylation has been reported in other cancers, a process that collaborates with its silencing and epithelial-mesenchymal tran sition of cervical. Conclusions. SFRP1 is a potential biomarker in preneoplastic lesions of the cervix associated with human papillomavirus.

Zebularine induces apoptosis of esophageal cancer cells via demethylation SFRP2/Dkk3 to regulate Wnt/β-catenin signaling pathway / 药学学报

To explore the effect and mechanisms of demethylation drug zebularine on esophageal cancer cells apoptosis, ECA109 cells and KYSE170 cells were treated with zebularine at different concentrations (25, 50, 100, 200, and 400 μmol·L-1). The cell viability was measured by CCK-8. Flow cytometry was used to detect the cell apoptosis rate, Western blot was performed to determine the expression of apoptosis protein (Bcl-2, Bax, cleaved-caspase-3, and cleaved-PARP) and Wnt signal pathway molecules (β-catenin, cyclin D1, and c-Myc), real-time quantitative PCR was used to detect the expression level of negative regulatory genes of Wnt signaling pathway, methylation specific PCR (MSP) was used to detect the methylation status of secreted frizzled related protein 2 (SFRP2) and dickkopf 3 (Dkk3) genes. After knockdown of SFRP2 and Dkk3, the effect of zebularine on apoptosis was detected. The studies showed that zebularine could inhibit the activity of ECA109 and KYSE170 cells in a dose-dependent and time-dependent manner; zebularine could induce cell apoptosis, down-regulate the expression of Bcl-2 protein, up-regulate the expression of Bax, cleaved-caspase-3, and cleaved-PARP protein, and inhibit the expression of β-catenin, cyclin D1, and c-Myc protein (P < 0.05); the mRNA expression levels of Dkk3 and SFRP2 were significantly up-regulated by zebularine, while the methylation levels of SFRP2 and Dkk3 promoters were decreased; knockdown of SFRP2 and Dkk3 could reduce the apoptosis induced by zebularine. In summary, zebularine could reduce the methylation level of SFRP2 and Dkk3 gene promoter, promote the expression of SFRP2 and Dkk3 gene, and then induce the apoptosis of esophageal cancer cells by inhibiting Wnt/β-catenin signaling pathway.

Association of serum levels of secreted frizzled-related protein 5 and Wnt member 5a with glomerular filtration rate in patients with type 2 diabetes mellitus and chronic renal disease: a cross-sectional study

ABSTRACT BACKGROUND: Diabetic nephropathy is a common complication of chronic kidney disease (CKD). ­Inflammation in the kidneys is crucial for promoting development and progression of this complication. Wnt member 5a (Wnt5a) and secreted frizzled-related protein 5 (Sfrp5) are proinflammatory proteins associated with insulin resistance and chronic low-grade adipose tissue inflammation. OBJECTIVE: To determine the correlation between serum Sfrp5 and Wnt5a concentrations and glomerular filtration rate in patients with type 2 diabetes mellitus and CKD. DESIGN AND SETTING: Cross-sectional, comparative and observational study in the Department of Endocrinology, Civil Hospital, Culiacán, Sinaloa, Mexico. METHODS: Eighty individuals with chronic kidney disease were recruited. Their serum Sfrp5 and Wnt5a concentrations were quantified using the enzyme-linked immunosorbent assay (ELISA) test. The statistical analysis consisted of the Mann-Whitney U test for independent samples and Spearman correlation, with statistical significance of P < 0.05. RESULTS: Serum Sfrp5 concentration continually increased through the stages of CKD progression, whereas serum Wnt5a concentration presented its highest levels in stage 3 CKD. Negative correlations between estimated glomerular filtration rate (eGFR) and serum concentrations of Sfrp5 (r = -0434, P = 0.001) and Wnt5a (r = -0481, P = 0.001) were found. CONCLUSIONS: There were negative correlations between serum Sfrp5 and Wnt5a concentrations and eGFR at each stage of CKD, with higher levels in female patients. This phenomenon suggests that Sfrp5 and Wnt5a might be involved in development and evolution towards end-stage renal disease.

Value of the methylation status of SDC2 and SFRP2 for colorectal cancer screening / 中华消化内镜杂志

Objective To explore the value of methylation of SDC2 and SFRP2 genes promoter in fecal DNA for colorectal cancer ( CRC) screening. Methods All stool samples were enrolled from Changhai Hospital of Naval Medical University, the Tenth People' s Hospital of Tongji University and the Seventh Medical Center of Chinese People's Liberation Army General Hospital. A total of 500 stool samples collected from March 2018 to December 2018 were allocated to CRC group ( 132 CRCs ) , adenoma group ( 38 advanced adenomas), healthy group (152 healthy individuals), interferential group (178 cases of benign colorectal disease or other non-colorectal tumors) and negative group (330 cases composed of healthy group and interferential group ) . The promoter methylation of fecal SDC2 and SFRP2 genes was detected by methylation-specific PCR (MSP) and compared with single gene methylation and the fecal immunochemical tests ( FIT) to evaluate its sensitivity and specificity. Results The stool sample analysis showed that the sensitivity of combined detection of SDC2 and SFRP2 in CRC group was 97. 73％ ( 129/132 ) , which was significantly higher than those of the single gene SDC2 test [ 70. 45％ ( 93/132) , P=0. 000] , single SFRP2 test [81. 82％ (108/132), P=0. 000] and FIT [69. 70％ (92/132), P=0. 000]. In adenoma group, the sensitivity of combined detection of SDC2 and SFRP2 was 57. 89％ (22/38), which was significantly higher than those of the single gene SDC2 test [ 15. 79％ ( 6/38 ) , P= 0. 000 ] and FIT [ 21. 05％ ( 8/38 ) , P=0. 021] , with no significant difference compared with that of SFRP2 test [ 47. 37％ ( 18/38) , P=0. 358] . In healthy group, the specificity of combined detection of SDC2 and SFRP2 was 98. 68％ (150/152), with no significant difference compared with those of single gene SDC2 test [ 100. 00％( 152/152) , P=0. 156] , single SFRP2 test [98. 68％ (150/152), P=1. 000] or FIT [95. 39％ (145/152), P=0. 091]. Specificities of combined detection of two genes in interferential and negative groups were 90. 45％ ( 161/178) and 94. 24％( 311/330) , which were significantly higher than 73. 03％( 130/178, P=0. 000) and 83. 33％( 275/330, P=0. 000) of FIT, respectively. Conclusion The combined detection test of methylation of SDC2 and SFRP2 is superior to single gene test, whose sensitivity of CRC and aggressive adenoma and specificity of distinguishing benign and malignant lesions are higher than FIT, which has potential application value.

MiR-1180 from bone marrow-derived mesenchymal stem cells induces glycolysis and chemoresistance in ovarian cancer cells by upregulating the Wnt signaling pathway / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

BACKGROUND@#Bone marrow-derived mesenchymal stem cells (BM-MSCs) play an important role in cancer development and progression. However, the mechanism by which they enhance the chemoresistance of ovarian cancer is unknown.@*METHODS@#Conditioned media of BM-MSCs (BM-MSC-CM) were analyzed using a technique based on microRNA arrays. The most highly expressed microRNAs were selected for testing their effects on glycolysis and chemoresistance in SKOV3 and COC1 ovarian cancer cells. The targeted gene and related signaling pathway were investigated using in silico analysis and in vitro cancer cell models. Kaplan-Merier survival analysis was performed on a population of 59 patients enrolled to analyze the clinical significance of microRNA findings in the prognosis of ovarian cancer.@*RESULTS@#MiR-1180 was the most abundant microRNA detected in BM-MSC-CM, which simultaneously induces glycolysis and chemoresistance (against cisplatin) in ovarian cancer cells. The secreted frizzled-related protein 1 (SFRP1) gene was identified as a major target of miR-1180. The overexpression of miR-1180 led to the activation of Wnt signaling and its downstream components, namely Wnt5a, β-catenin, c-Myc, and CyclinD1, which are responsible for glycolysis-induced chemoresistance. The miR-1180 level was inversely correlated with SFRP1 mRNA expression in ovarian cancer tissue. The overexpressed miR-1180 was associated with a poor prognosis for the long-term (96-month) survival of ovarian cancer patients.@*CONCLUSIONS@#BM-MSCs enhance the chemoresistance of ovarian cancer by releasing miR-1180. The released miR-1180 activates the Wnt signaling pathway in cancer cells by targeting SFRP1. The enhanced Wnt signaling upregulates the glycolytic level (i.e. Warburg effect), which reinforces the chemoresistance property of ovarian cancer cells.

MiR-1180 from bone marrow-derived mesenchymal stem cells induces glycolysis and chemoresistance in ovarian cancer cells by upregulating the Wnt signaling pathway / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Background: Bone marrow-derived mesenchymal stem cells (BM-MSCs) play an important role in cancer development and progression. However, the mechanism by which they enhance the chemoresistance of ovarian cancer is unknown. Methods: Conditioned media of BM-MSCs (BM-MSC-CM) were analyzed using a technique based on microRNA arrays. The most highly expressed microRNAs were selected for testing their effects on glycolysis and chemoresistance in SKOV3 and COC1 ovarian cancer cells. The targeted gene and related signaling pathway were investigated using in silico analysis and in vitro cancer cell models. Kaplan-Merier survival analysis was performed on a population of 59 patients enrolled to analyze the clinical significance of microRNA findings in the prognosis of ovarian cancer. Results: MiR-1180 was the most abundant microRNA detected in BM-MSC-CM, which simultaneously induces glycolysis and chemoresistance (against cisplatin) in ovarian cancer cells. The secreted frizzled-related protein 1 (SFRP1) gene was identified as a major target of miR-1180. The overexpression of miR-1180 led to the activation of Wnt signaling and its downstream components, namely Wnt5a, β-catenin, c-Myc, and CyclinD1, which are responsible for glycolysis-induced chemoresistance. The miR-1180 level was inversely correlated with SFRP1 mRNA expression in ovarian cancer tissue. The overexpressed miR-1180 was associated with a poor prognosis for the long-term (96-month) survival of ovarian cancer patients. Conclusions: BM-MSCs enhance the chemoresistance of ovarian cancer by releasing miR-1180. The released miR-1180 activates the Wnt signaling pathway in cancer cells by targeting SFRP1. The enhanced Wnt signaling upregulates the glycolytic level (i.e. Warburg effect), which reinforces the chemoresistance property of ovarian cancer cells.

Expression of WNT4 and its inhibitory factor SFRP1 in renal tissue of diabetic nephropathy rats / 基础医学与临床

Objective To observe the change of expression of WNT4/β-catenin signaling pathway and its inhibitory factor secreted frizzled-related protein 1 (SFRP1) in renal tissue in diabetic nephropathy(DN) rats, and to explore its possible role in the development of renal fibrosis. Methods Rats were randomly divided into normal control(NC) group and DN group, and equipped with 8 in each group. The IDDM model was prepared by tail vein injection of STZ 55 mg/kg. Hemotoxyin and eosin、Periodic Acid-Schiff and Masson stain were used to observe the morphological structure and fibrotic lesions in renal tissue;Immunohistochemical analysis was used to observe the protein expression of WNT4 and β-catenin in renal tissue;Western blot was used to detect the protein expression changes of WNT4, SFRP1, β-catenin, p-GSK-3β, GSK-3β, Collagenl, a-SMA, E-cadherin in renal tissue in each group;The mRNA expression of WNT4 and SFRPl in renal tissues of rat was detected by realtime PCR. Results Compared with NC group, renal tissue fibrosis was obvious in DN group. Compared with NC group, the protein and mRNA expressions of WNT4 significantly increased (P＜0.05), the protein expressions of β-catenin, p-GSK-3β, α-SMA and collagen I significantly increased (P < 0.05), the protein expressions of Ecadherin significantly decreased (P＜0. 05), the protein and mRNA expression of SFRPl significantly decreased (P＜0.05). Conclusions In the case of DN, the signal pathway of WNT4/β-catenin is abnormal activation. The expression of SFRPl is decreased, and that may inhibit this pathway and promote the development of renal fibrosis in DN.

Study on inhibition role of SFRP2 on cervical cancer cell line proliferation / 重庆医学

Objective To understand the expression change of SFRP2 in human cervical cancer tissue and to investigate the effect of SFRP2 on cervical cancer cell proliferation.Methods The expression of SFRP2 in cervical cancer tissue was detected by using Western blot and qRT-PCR;the SFRP overexpressed human cervical cancer line was constructed by using lentivirus,the effect of SFRP2 on the proliferation of human cervical cancer cell line was analyzed by CCK-8 and plate cloning.The effect of SFRP2 on the expression of WNT pathway related proteins and genes in human cervical cancer cell was detected by Western Bolt and qRTPCR.Results Compared with paracancerous tissue,SFRP2 was lowly expressed in human cervical cancer tissue;overexpressed SFRP2 cervical cancer cell proliferation was inhibited;SFRP2 inhibiting cellular proliferation was occurred via WNT signal pathway.Conclusion The role of SFRP2 as a candidate gene for cervical cancer remains to be deeply studied.

Relationship between plasma secreted frizzled-related protein 5 with type 2 diabetes mellitus and carotid atherosclerosis / 中国糖尿病杂志

Objective To investigate the relationships between plasma secreted frizzled-related protein (SFRP) 5 level with type 2 diabetes (T2DM ) and carotid atherosclerosis (CAS). Methods According to the results of carotid color Doppler ultrasound ,56 T2DM patients were divided into 28 with carotid atherosclerosis (CAS group) and 28 without carotid atherosclerosis (NAS group). 28 healthy volunteers served as normal control (NC group). Serum SFRP5 was measured by ELISA. Results The level of serum SFRP5 in T2DM patients was lower than NC group (114.36 ± 25.48) vs (48.19 ± 11.82) , (43.88 ± 8.19)pg/ml ,(P 0.05 ]. Pearson correlation analysis showed that serum SFRP5 was negatively correlated with age ,TG ,hsC-RP ,FPG ,FIns , HOMA-IR ,HbA1c ,TC ,LDL-C and BMI (P0.05). Multiple linear regression results showed that age ,FIns and HbA1 c were independent influencing factors of serum SFRP5. Conclusion SFRP5 may be a protective factor for T2DM by ameliorating insulin resistance which may provide a new clue for the prevention and treatment of T 2DM.

Correlation between Hypermethylation of SFRP2 promoter CpG island and colorectal cancer / 中国现代普通外科进展

Objective:SFRP2 gene is a member of the SFRPs family.The gene is located on chromosome 4q31.3 with 3 exons and 2 introns and first exons have higher density near the island of CpG.Many studies showed that the methylation level of SFRP2 gene and colon cancer,esophageal cancer,gastric cancer and other tumor occurrence relating to,development and prog nosis.This study aims to study the clinical characteristics of CpG SFRP2 promoter island hypermethylation in colorectal cancer and whether there is a certain correlation.Methods:by matrix assisted laser desorption ionization time of flight mass spectrometry for detecting specific CpG island methylation.Methylation status of SFRP2 promoter by Sequenom EpiTYPER was detected in 20 cases of normal tissue of colorectal cancer and tumor tissues.Results:Our study using multiple linear regression analysis in tumor tissue of SFRP2 methylation at promoter Ⅰ and Ⅱ,found that SFRP2 promoter methylation and clinical features of.SFRP2_01_CpG_5 significantly correlated (P=0.018),SFRP2_02_CpG_5 (P=0.018) associated with the location of the tumor,SFRP2_02_CpG_6,7,8,9(P=0.039) and the number of lymph node metastasis of.SFRP2_01_CpG_1.2(P=0.043),SFRP2_02_CpG_16 (P=0.044) correlated with tumor size.Conclusion:we from epigenetic aspects of its promoter CpG methylation level and colorectal cancer clinical and pathological features,found a correlation between clinical and pathological features of colorectal cancer and CpG methylation in its promoter,suggesting that the SFRP2 promoter may be at this stage of colorectal cancer and future biological genetics the progress of the potential surface markers.

Alteration of Wnt/β-catenin signaling pathway in type2 diabetic rats’ aorta and regulation of SIRT1 / 中国药理学通报

Aim To investigate the alteration of Wnt/β-catenin signaling and sirtuins 1 in type 2 diabetic rats’ aorta and clarify its role in the development of di-abetes aortic disease. Methods The type 2 diabetes rat model was established by injection of streptozocin after five-week of high fat diet. The rats were randomly divided into control group, DM model group of 2 weeks, 4 weeks, 8 weeks and 12 weeks. Fasting blood glucose ( FBG ) , total cholesterol ( TC ) , triglyceride ( TG) , high density lipoprotein-cholesterol( HDL-C) , low density lipoprotein- cholesterol ( LDL-C ) and fast-ing insulin( FINS) levels were tested. HE staining was used to observe the pathological changes of aortal struc-tures. The alteration of Wnt2, β-catenin, TCF4, SIRT1 and sFRP2 in aortawas determined by Western blot and RT-PCR. Results Compared with control group, TC, TG, LDL-C levels of type 2 diabetic rats were significantly increased, HDL-C levels were signif-icantly reduced( P0. 05). But the expression of TCF4 and SIRT1 was enhanced continuously in DM compared with control group while sFRP2 decreased in the duration of DM development. Conclusions Wnt/β-catenin signa-ling pathway was activated in diabetic aortal injury by regulation of SIRT1 via sFRP2 . Further researches on its mechanism of actionin DM aorta injury may find a new therapeutic target for the disease.

Relationship between Sfrp5 and T2DM macroangiopathy and activation of calcium dobesilate / 实用医学杂志

Objective To investigate the relationship between Sfrp5 and T2DM macroangiopathy , and to evaluate the activation of calcium dobesilate. Methods T2DM patients were divided into experimental group (IMT≥0.90 mm,n = 65)and control group (IMT 0.05). IMT in group A were decreased when compared with group B (P < 0.05). Conclusion Sfrp5 is negatively correlated with macroangiopathy. Calcium dobesilate can elevate Sfrp5 and IL-10,thus delay the progression of macroangiopathy.

Methylation of SFRP1 gene promoter in cutaneous squamous cell carcinoma tissue / 中华皮肤科杂志

Objective To investigate the relationship between SFRP1 gene and clinicopathologic features of cutaneous squamous cell carcinoma (CSCC), and to explore the possible mechanism of action of SFRP1 in the occurrence and development of CSCC.Methods CSCC and paracarcinomatous tissue specimens were obtained from 40 patients with CSCC, and normal skin tissue specimens from 40 healthy human controls.The EpiTYPER assay was conducted to evaluate the methylation status of SFRP1 gene promoter in all the specimens with a MassARRAY mass spectrometer.Results Totally, the methylation status of 1951 (86.52％, 1951/2255) CpG motifs were evaluated in 17 CpG loci in 2 fragments of the SFRP1 gene promoter.The methylation rate significantly differed in 10 (10/17, 58.82％) CpG loci between the CSCC and paracarcinomatous tissue specimens, and in 5 (5/17, 29.41％) CpG loci between the paracarcinomatous and normal tissue specimens (all P ＜ 0.05).Furthermore, significant differences were observed in the methylation rates of three CpG loci (CpG 1_5, CpG 1_7, CpG 2_8) in the SFRP1 gene promoter between tissue specimens from different pathological grades of CSCC (P ＜ 0.05), and their methylation rates sequentially decreased from grade Ⅲ to grade Ⅱ and Ⅰ.Conclusion The frequency of methylation is high in the SFRP1 gene promoter in patients with CSCC, and the SFRP1 gene may participate in the occurrence and development of CSCC.

Expression of Sfrp2 Is Increased in Catagen of Hair Follicles and Inhibits Keratinocyte Proliferation

BACKGROUND: Hair follicles undergo cycles of repeated growth and regression. The Wnt pathway plays an important role in the regeneration and differentiation of hair follicles. Sfrp2, a Wnt inhibitor, is involved in the developmental and disease processes of various cells and tissues by modulating the Wnt pathway. OBJECTIVE: The aim of this study was to understand the role of Sfrp2 in hair follicles through investigation of the Sfrp2 expression pattern in the skin and its effect on keratinocytes. METHODS: We investigated Sfrp2 mRNA expression and the expression of the wnt target genes, Ccnd1 and C-myc, at various mouse hair follicle developmental stages using Real-time polymerase chain reaction. We also investigated the effect of SFRP2 on the proliferation and differentiation of mouse keratinocyte cells by adding SFRP2 protein or overexpressing Sfrp2 using an in vitro culture system. RESULTS: Sfrp2 expression peaked in the catagen phase and remained high until telogen, and then declined at the beginning of the next anagen. An inverse relationship to Sfrp2 expression was found for the expression of the Wnt target genes, C-myc and Ccnd1. In addition, we also observed inhibited proliferation of mouse keratinocytes in the presence of SFRP2. CONCLUSION: These results suggest that Sfrp2 may play a role in the catagen phase by inhibiting the proliferation of keratinocyte and functioning as a Wnt inhibitor in keratinocytes.

The Expression and Clinical Significance of DKK1, SFRP4 and Wnt1 in Cervical Squamous Cell Carcinoma / 天津医药

Objective To investigate the expressions of DKK1,SFRP4 and Wnt1 in cervical squamous cell carcino-ma(SCC), and the clinical significance thereof. Methods There were 76 samples of cervical squamous cell carcinoma were included in SCC group and 36 benign uterine resection specimens were control group (NC). The immunohistochemical meth-od was applied to detect the expressions of DKK1,SFRP4 and Wnt1 in two groups. Results The expression of DKK1 was significantly lower in SCC group than that in NC group (P＜0.05). The expression levels of SFRP4 and Wnt1 were significant-ly higher in SCC group than those of NC group (P＜0.05). There were significant differences in the expressions of DKK1, SFRP4 and Wnt1 between samples of different clinical staging, differentiation, sizes of tumor and lymph node metastasis (P＜0.05). The expression of DKK1 was negatively correlated with SFRP4 and Wnt1 in SCC group (P＜0.05). The expression of SFRP4 was positively correlated with Wnt1 in SCC group (P＜0.05). Conclusion The roles of SFRP4 and Wnt1 are syn-ergistic interactions in the development of SCC. DKK1 is an inhibiting factor of SCC.

Chuju total flavonoids control the SFRP4 expression in Wnt pathway in rheumatoid arthritis model rats / 中南大学学报(医学版)

Objective:To determine the effect of Chuju total flavonoids (CJTF) on the secreted frizzled-related protein 4 (SFRP4) expression in Wnt pathway in rheumatoid arthritis (AR) model rats. Methods:hTe role of CJTF in the treatment of AR model rats was evaluated by rat arthritis score and paw edema score. The expression regulation of the SFRP4,β-catenin and C-myc in Wnt pathway in AR model rats was detected by RT-PCR and Western blot atfer CJTF gavage treatment. Results:Atfer CJTF treatment, the rat arthritis score and paw edema score in AR model rats were signiifcantly decreased when the AR model rats were treated with CJTF, the SFRP4 expression was signiifcantly up-regulated, while theβ-catenin and C-myc gene expression were signiifcantly down-regulated in AR model rat synovial tissues. Conclusion:CJTF has significant therapeutic effect and inhibitory effect on Wnt pathway activation by targeting SFRP4 in AR model rat synovium.

The Association of Polymorphisms of Secreted Frizzled Related Protein Genes with Circulating Osteoprotegerin (OPG): Soluble Receptor Activator of NF-kB Ligand (sRANKL) and Bone Mineral Density / 대한골다공증학회지

OBJECTIVES: To investigate the relationship between polymorphisms of secreted frizzled related protein (sFRP) genes, circulating osteoprotegerin (OPG), and soluble receptor activator of NF-kB ligand (sRANKL), and bone mineral density (BMD) in postmenopausal Korean women. METHODS: The sFRP1 c.3132C>T, rs16890444, sFRP2 c.-38C>G, and sFRP5 c.20G>C, and c.34A>T polymorphisms were analyzed by Taqman assay and dierect DNA sequencing in 170 postmenopausal Korean women. Serum CrossLaps (CTX), bone alkaline phosphatase (BAP), OPG, and sRANKL were measured by enzyme linked immunosorbent assay. The BMD at the lumbar spine and femoral neck was determined by dual energy X-ray absorptiometry. RESULTS: The sFRP2 c.-38C>G, and sFRP5 c.34A>T polymorphisms were not observed. No significant differences in adjusted BMD of lumbar spine and femoral neck and in risk for osteoporosis were noted among single genotypes of the sFRP genes polymorphisms measured or haplotype genotypes composed of the sFRP1 c.3132C>T and rs16890444 polymorphisms. No statistical significances in serum levels of bone chemical markers except BAP were observed according to single or haplotype genotypes. Serum BAP was significantly higher in women with the GG genotype than those in women with the GC genotype of the sFRP5 c.20G>C polymorphism. CONCLUSIONS: The sFRP1 c.3132C>T, rs16890444 and sFRP5 c.20G>C polymorphisms do not affect BMD at the lumbar spine and femoral neck in postmenopausal Korean women.

Regulation of the Wnt Signaling Pathways during Cell Culture of Human Mesenchymal Stem Cells for Efficient Bone Regeneration / Oral Science International

Tissue engineering and bone regeneration techniques using mesenchymal stem cells (MSCs) have started to be applied to the field of oral and maxillofacial surgery. Clinically, a shortened treatment time and improved efficiency are necessary because of the patients' needs and the running cost of cell culture. In the present study, the cultivation process for human MSCs (hMSCs) was examined by regulating the Wnt signaling pathway. We activated Wnt signaling with LiCl and inhibited Wnt signaling with sFRP-3 (secreted Frizzled-Related Protein-3). The proliferation of LiCl-treated hMSCs was examined by studying the cell growth rate and performing BrdU assays. Osteogenic differentiation of sFRP-3-treated hMSCs was examined by alizarin red staining, and osteogenic gene expression on days 7 and 14 after induction was examined by reverse-transcription polymerase chain reaction (RT-PCR) analysis and quantitative real-time RT-PCR analysis. LiCl-treated hMSCs showed increased cell numbers and BrdU-positive cells as compared to the untreated cells. Alizarin red staining showed early mineralization of hMSCs on day 7 of the sFRP-3 treatment. A high expression level of the alkaline phosphatase gene on days 7 and 14 of sFRP-3 treatment was also demonstrated. These results suggest that the regulation of the Wnt signaling pathway contributes to the increased cell numbers and the early osteogenic differentiation of hMSCs. This study supports the possibility that the regulation of the Wnt signaling pathway contributes to the development of effective and efficient bone regeneration techniques.

Aberrant promoter methylation of SFRP1 and SFRP2 gene in gastric cardia adenocarcinoma / 肿瘤研究与临床

Objective To investigate the promoter methylation status of SFRP1 and SFRP2 gene in gastric cardia adenocarcinoma (GCA). Methods Methylation specific PCR (MSP) method was used to examine the methylation status of the 5' CpG island of SFRP1 and SFRP2 gene in tumors and corresponding normal tissues. Results Methylation frequencies of SFRP1 and SFRP2 gene in tumor specimens were 87.2 % (82/94) and 83 %(78/94), which was significantly higher than that in corresponding normal tissues (14.9 % and 55.3 %, respectively) (P <0.001). Methylation frequencies of SFRP1 in lymph node metastasis group (96.4 %) was significantly higher than that in no lymph node metastasis group (73.7 %). Methylation frequencies of SFRP1 and SFRP2 gene in poor differentiation group were all higher than that in moderate and poor-moderate differentiation groups, but both of them did not show significant difference(P >0.05). 63 cases of GCA showed both of SFRP1 and SFRP2 gene simultaneous methylation, which including 36 cases of lymph node metastasis group, 27 cases of no lymph node metastasis group. Simultaneous methylation frequencies of SFRP1 and SFRP2 gene in lymph node metastasis group was higher than that in no lymph node metastasis group, poor differentiation group was higher than that in moderate and poor-moderate differentiation groups, but both of them did not show significant difference (P >0.05). Conclusion Promoter methylation of SFRP1 and SFRP2 might be related with oncogenesis of GCA and hypermethylation of SFRP1 gene might be related with the malignant behavior of GCA.