Establishment and identification of secreted frizzled-related protein 4 transgenic mice and analysis of their biological properties / 西安交通大学学报(医学版)

【Objective】 To generate secreted frizzled-related protein 4 (SFRP4) transgenic mice and analyze their biological properties. 【Methods】 We generated SFRP4 transgenic mice by DNA microinjection and detected the expression of SFRP4 by qRT-PCR and Western blotting. 【Results】 SFRP4 transgenic mice were successfully generated by DNA microinjection. The expression of SFRP4 was dramatically increased in transgenic mice liver compared with that of wide type. 【Conclusion】 The transgenic mice model of SFRP4 was successfully established by DNA microinjection. It provides a good model for studying the function of SFRP4 and the mechanism of participating in metabolic diseases.

Expression of secreted frizzled-related protein 4 in DNA mismatch repair-deficient and mismatch repair-proficient colorectal cancers / 南方医科大学学报

OBJECTIVE@#To investigate the expressions of secreted frizzled-related protein 4 (SFRP4) in stage Ⅱ DNA mismatch repair-deficient (dMMR) and mismatch repair- proficient (pMMR) colorectal cancers and explore their clinical significance.@*METHODS@#We collected fresh stage Ⅱ colon cancer tissues with different MMR status detected by immunohistochemistry (IHC). The differentially expressed mRNAs between dMMR and pMMR tumors were identified by Affymetrix Human oeLncRNA gene chip, and the expression of SFRP4 in these cancer tissues and in colorectal cancer cell lines were detected using Western blotting and real- time quantitative PCR. The apoptosis rates of HCT116 cells with and without siRNA- mediated transient SFRP4 knockdown were determined using flow cytometry. We further investigated the expression pattern of Ki-67 and its correlation with SFRP4 expression.@*RESULTS@#Compared with pMMR colon cancer tissues or cells, both dMMR colon cancer tissues (=0.014) and cells (=0.0079) showed significantly increased expression of SFRP4, which was in negative correlation with Ki-67 (=0.041). In HCT116 cells, transient SFRP4 knockdown resulted in decreased cell apoptosis, including both early apoptosis (=0.003) and late apoptosis (=0.024).@*CONCLUSIONS@#Up-regulation of SFRP4 in dMMR stage Ⅱ colon cancer promotes apoptosis and inhibits proliferation of the cancer cells, and may improve the prognosis of dMMR colon cancer.

Expression and clinical significance of SFRP4 in pancreatic ductal adenocarcinoma / 临床与实验病理学杂志

Purpose To investigate the expression of secreted frizzled-related protein 4 (SFRP4) and to evaluate its clinical significance in pancreatic ductal adenocarcinoma (PDAC).Methods RT-PCR was performed to analyze SFRP4 mRNA expression level in 30 paired PDAC lesion and matched adjacent non-tuimor tissue.Immunohistochemistry staining detection of 205 matched cases tissue microarray was conducted to explore SFRP4 protein expression pattern.The correlation between SFRP4 and clinical characteristics was also analyzed,including overall survival.Results SFRP4 expression pattern both at mRNA and protein level in PDAC lesion was higher than that in matched adjacent non-tumor tissue.At mRNA level,to found that expression of SFRP4 was elevated in 90％ (27/30) of PDAC tissues (P =0.007 2).To found that high expression of SFRP4 was detected in 56.5％ (116/205) of PDAC tissue,while only 28.8％ (59/205) in the adjacent non-tumor tissue.Moreover,no significant association was observed between SFRP4 expression and clinical characteristics.Kaplan-Meier survival analysis showed high level of SFRP4 expression was correlated with poor overall survival (x2 =3.467,P =0.024).Conclusion SFRP4 can be a novel prognostic biomarker in PDAC.

Relationship between serum secreted Frizzled-related protein 4 and the pancreaticβcell function / 中华内分泌代谢杂志

Objective To investigate the relationship between serum secreted frizzled-related protein 4 (SFRP4) and the first-phase of glucose-stimulated insulin secretion from pancreatic β cell under different glucose tolerance statuses. Methods Fifty-six patients with newly diagnosed type 2 diabetes mellitus ( T2DM group), 52 patients with impaired glucose tolerance (IGT group), and 42 subjects with normal glucose tolerance (NGT group) underwent intravenous glucose tolerance test. Fasting serum SFRP4 and interleukin ( IL)-1β were assayed by ELISA. Acute insulin response ( AIR), the area under the curve of the first-phase (0-10 min) insulin secretion (AUC), glucose disposition index(GDI), homeostasis model assessment for β cell function index(HOMA-β), and insulin resistance index(HOMA-IR) were calculated. Results (1) The levels of SFRP4 and IL-1β in T2DM group and IGT group were significantly higher than that in NGT group [(184. 38 ± 61. 34 or 141. 64 ± 40. 46 or 95. 46 ± 20. 13)ng/ ml, P<0. 01]. AIR, AUC, and GDI in T2DM group and IGT group were significantly lower than those in NGT group(P<0. 01), and these results were more significantly reduced in T2DM group compared with those in IGT group. (2) SFRP4 was negatively correlated with AIR, AUC, GDI, HOMA-β (P<0. 01), and positively correlated with fasting plasma glucose, 2 h plasma glucose after glucose loading, HbA1C , IL-1β, and high sensitive C-reactive protein(P<0. 01). (3) Multiple stepwise regression analysis showed that AUC, HOMA-IR, and serum IL-1β level were independently associated with SFRP4. Conclusion The concentration of serum SFRP4 is closely correlated with the glycolipid metabolic disorder, the first-phase of glucose-stimulated insulin secretion, and chronic low-grade inflammation. SFRP4 may be involved in the mechanism of β cell dysfunction in type 2 diabetes mellitus.

Effects of magnolol in combination with 5-FU on cell proliferation and SFRP-4 expression of human colon cancer HCT-8 cells / 肿瘤

Objective: To investigate the effects of magnolol in combination with 5-fluorouracil (5-FU) on the proliferation of human colon cancer HCT-8 cells and the expression of secreted frizzled-related protein-4 (SFRP-4). Methods: After treatment with magnolol and 5-FU alone or magnolol in combination with 5-FU, the proliferation of colon cancer HCT-8 cells was detected by MTT method, and the cell cycle distribution and the apoptosis of HCT-8 cells were examined by flow cytometry (FCM) and Hoechst 33258 nucleic acid staining, respectively. The expression levels of SFRP-4 and p-catenin proteins in HCT-8 cells were examined by Western blotting. Results: The proliferation of HCT-8 cells was inhibited by magnolol and 5-FU alone or mamgnolol in combination with 5-FU (all P 2/M phases were decreased in 5-FU alone group and the combination treatment group (both P < 0.05). The expression levels of SFRP-4 in HCT-8 cells in all three treatment groups were higher than that in the blank control group (without any treatment) (P < 0.05), while the expression level of p-catenin in HCT-8 cells in combination treatment group was lower than those in 5-FU and magnolol alone groups (P < 0.05). Conclusion: Magnolol in combination with 5-FU can enhance the inhibition effect on proliferation of human colon cancer HCT-8 cells, and arrest the cells at G0/G1 phase. This mechanism may be related to regulating the expressions of SFRP-4 and p-catenin proteins.