AIM： To investigate the expression and methylation status of the secreted frizzled-related protein 2 （SFRP2） in esophageal squamous cell carcinoma （ESCC） and ex- plore its role in ESCC carcinogenesis.METHODS： Se...AIM： To investigate the expression and methylation status of the secreted frizzled-related protein 2 （SFRP2） in esophageal squamous cell carcinoma （ESCC） and ex- plore its role in ESCC carcinogenesis.METHODS： Seven ESCC cell lines （KYSE 30, KYSE150, KYSE410, KYSE510, EC109, EC9706 and TE-1） and one immortalized human esophageal epithelial cell line （Het- 1A）, 20 ESCC tissue samples and 20 paired adjacent non-tumor esophageal epithelial tissues were analyzed in this study. Reverse-transcription polymerase chain reaction （RT-PCR） was employed to investigate the expression of SFRP2 in cell lines, primary ESCC tumor tissue, and paired adjacent normal tissue. Methylation status was evaluated by methylation-specific PCR and bisulfite sequencing. The correlation between expres- sion and promoter methylation of the SFRP2 gene was confirmed with treatment of 5-aza-2＇-deoxycytidine. To assess the potential role of SFRP2 in ESCC, we es-tablished stable SFRP2-transfected cells and examined them with regard to cell proliferation, colony formation, apoptosis and cell cycle in vivo and in vitro.RESULTS： SFRP2 mRNA was expressed in the im- mortalized normal esophageal epithelial cell line but not in seven ESCC cell lines. By methylation-specific PCR, complete methylation was detected in three cell lines with silenced SFRP2 expression, and extensive methylation was observed in the other four ESCC cell lines. 5-aza-2＇-deoxycytidine could restore the expres- sion of SFRP2 mRNA in the three ESCC cell lines lack- ing SFRP2 expression. SFRP2 mRNA expression was obviously lower in primary ESCC tissue than in adjacent normal tissue （0.939 ± 0.398 vs 1.51 ± 0.399, P 〈 0.01）. SFRP2 methylation was higher in tumor tissue than in paired normal tissue （95% vs 65%, P 〈 0.05）. The DNA methylation status of the SFRP2 correlated inversely with the SFRP2 expression. To assess the potential role of SFRP2 in ESCC, we established stable SFRP2 transfectants and control counterparts by in- troducing pcDNA3.1/v5 hisA -SFRP2 or pcDNA3.1/v5 hisA -empty vector into KYSE30 cells lacking SFRP2 expression. After transfection, the forced-expression of SFRP2 was confirmed by the RT-PCR. In comparison with the control groups, stably-expressed SFRP2 in KYSE 30 cells significantly reduced colony formation in vitro （47.17% 4± 15.61% vs 17% ：1： 3.6%, P = 0.031） and tumor growth in nude mice （917.86：1：249.35 mm3 vs 337.23 ± 124.43 mm3, P 〈 0.05）. Using flow cytom- etry analysis, we found a significantly higher number of early apoptotic cells in SFRP2-transfected cells than in the control cells （P = 0.025）. The mean cell number in the S and G2-M phases of the cell cycle was also significantly lower in SFRP2-transfected KYSE30 cells compared with mock transfected counterparts. CONCLUSION： Silencing of SFRP2 expression through promoter hypermethylation may be a factor in ESCC carcinogenesis through loss of its tumor-suppressive activity.展开更多
Objective Microcystin-leucine-arginine(MC-LR)exposure induces lipid metabolism disorders in the liver.Secreted frizzled-related protein 5(SFRP5)is a natural antagonist of winglesstype MMTV integration site family,memb...Objective Microcystin-leucine-arginine(MC-LR)exposure induces lipid metabolism disorders in the liver.Secreted frizzled-related protein 5(SFRP5)is a natural antagonist of winglesstype MMTV integration site family,member 5A(Wnt5a)and an anti-inflammatory adipocytokine.In this study,we aimed to investigate whether MC-LR can induce lipid metabolism disorders in hepatocytes and whether SFRP5,which has anti-inflammatory effects,can alleviate the effects of hepatic lipid metabolism by inhibiting the Wnt5a/Jun N-terminal kinase(JNK)pathway.Methods We exposed mice to MC-LR in vivo to induce liver lipid metabolism disorders.Subsequently,mouse hepatocytes that overexpressed SFRP5 or did not express SFRP5 were exposed to MC-LR,and the effects of SFRP5 overexpression on inflammation and Wnt5a/JNK activation by MC-LR were observed.Results MC-LR exposure induced liver lipid metabolism disorders in mice and significantly decreased SFRP5 mRNA and protein levels in a concentration-dependent manner.SFRP5 overexpression in AML12cells suppressed MC-LR-induced inflammation.Overexpression of SFRP5 also inhibited Wnt5a and phosphorylation of JNK.Conclusion MC-LR can induce lipid metabolism disorders in mice,and SFRP5 can attenuate lipid metabolism disorders in the mouse liver by inhibiting Wnt5a/JNK signaling.展开更多
AIM： To investigate the functions of promoter hypermethylation of secreted frizzled-related proteins （sFRPs） genes in colorectal tumorigenesis and progression. METHODS： The promoter hypermethylation and expression...AIM： To investigate the functions of promoter hypermethylation of secreted frizzled-related proteins （sFRPs） genes in colorectal tumorigenesis and progression. METHODS： The promoter hypermethylation and expression of sFRP genes in 72 sporadic colorectal carcinomas, 33 adenomas, 18 aberrant crypt foci （ACF） and colorectal cancer cell lines RKO, HCT116 and SW480 were detected by methylation-specific PCR and reverse transcription PCR, respectively. RESULTS： None of the normal colorectal mucosa tissues showed methylated bands of any of four sFRP genes, sFRP1, 2, 4 and 5 were frequently methylated in colorectal carcinoma, adenoma and ACF （sFRP1 〉 85%, sFRP2 〉75%, sFRP5 〉 50%）, and the differences between three colorectal tissues were not significant （P 〉 0.05）. IVlethylation in colorectal tumors was more frequent than in normal mucosa and adjacent normal mucosa. The mRNA of sFRP1-5 genes was expressed in all normal colorectal mucosa samples. Expression of sFRP1, 2, 4 and 5 and sFRP1, 2 and 5 was downregulated in carcinoma and adenoma, respectively. The downregulation of sFRP2, 4 and 5 was more frequent in carcinoma than in adenoma. Expression of sFRP3 which promoter has no CpG island was downregulated in only a few of colorectal tumor samples （7/105）. The downregulation ofsFRP1, 2, 4 and 5 expression was significantly associated with promoter hypermethylation in colorectal tumor. After cells were treated by DAC/TSA combination, the silenced sFRP mRNA expression could be effectively re-expressed in colorectal cancer cell lines. CONCLUSION： Hypermethylation of sFRP genes is a common early event in the evolution of colorectal tumor, occurring frequently in ACF, which is regarded as the earliest lesion of multistage colorectal carcinogenesis. It appears to functionally silence sFRP genes expression. Methylation of sFRP1, 2 and 5 genes might serve as indicators for colorectal tumor.展开更多
AIM： To identify the methylation of secreted frizzled-related protein 1 （SFRP1） in gastric cancer and to investigate the aberrant expression of SFRP1 and its correlation with the clinical pathological features of p...AIM： To identify the methylation of secreted frizzled-related protein 1 （SFRP1） in gastric cancer and to investigate the aberrant expression of SFRP1 and its correlation with the clinical pathological features of patients. METHODS： We determined SFRP1 methylation and SFRP1 mRNA expression in 3 gastric cancer cell lines SGC-7901, BGC-823, HGC-27, from 52 primary gastric cancer specimens and matched tumor adjacent tissue specimens by methylation-specific （MSP） PCR and RT-PCR respectively. Fisher＇s exact test was used to analyze the statistical association between clinical pathological data and aberrant expression of SFRP1. RESULTS： In 3 cancer cell lines, BGC-823 and HGC-27 had methylated SFRP1 and lost SFRP1 mRNA expression. After treatment of BGC-823 and HGC-27 with the demethylating agent, 5-aza-2′-deoxycytidine, SFRP1 was re-expressed. In 52 primary gastric cancer specimens and matched tumor adjacent tissue specimens, hypermethylation of SFRP1 was detected in 23 （44%） and 8 （15%） specimens respectively （x^2= 10.34, P 〈 0.01）. Loss of SFRP1 expression was detected in 17（33%） and 6 （12%） specimens respectively （x^2= 6.75, P 〈 0.01）. There was a significant correlation between SFRP1 hypermethylation and SFRP1 expression loss. SFRP1 expression was also correlated significantly with tumor stage and lymph node status, but not with patient sex, age and histological type. CONCLUSION： SFRP1 inactivation is a common and early event caused mainly by hypermethylation in gastric cancer. SFRP1 expression loss may be correlated with tumor metastasis in primary gastric cancer.展开更多
Objective Obstructive sleep apnea (OSA) is closely related to obesity, insulin resistance and inflammation. Secreted frizzled-related protein 5 (SFRP5) is a recently discovered adipokine. It is involved in insulin res...Objective Obstructive sleep apnea (OSA) is closely related to obesity, insulin resistance and inflammation. Secreted frizzled-related protein 5 (SFRP5) is a recently discovered adipokine. It is involved in insulin resistance and inflammation in obesity. This study aimed at evaluating the association between SFRP5and sleeping characteristics as well as biochemical parameters of OSA patients.Methods This was a prospective case control study. Nondiabetic OSA patients and controls were consecutively recruited and divided into three groups: OSA group, apnea–hypopnea Index (AHI)≥5/h; healthy controls with normal body mass index (BMI); obese controls without OSA, and BMI > 24.0 kg/m2. All participants underwent polysomnography (PSG). Plasma SFRP5 was examined using enzyme-linked immunosorbent assay (ELISA). Blood biochemical examinations, including fasting blood glucose (FBG), lipid profile, hypersensitive Creactive protein (hsCRP), were performed early in the morning after PSG. Patients with severe OSA were treated with nasal continuous positive airway pressure (nCPAP), and plasma SFRP5 was repeatedly measured for comparison.Results Sixty-eight subjects were enrolled in the study, including 38 patients of OSA, whose medium AHI was 58.70 /h (36.63, 71.15), 20 obese controls, and 10 healthy controls. The plasma SFRP5 level of OSA patients was not significantly different from that of healthy controls or obese controls. In OSA patients, SFRP5 level correlated positively with triglyceride level (r=0.447, P=0.005) and negatively with LDL-cholesterol level and HDLcholesterol level (r=?0.472 and P=0.003; r=?0.478 and P=0.002; respectively). SFRP5 level was not found correlating with FBG, AHI, or any of nocturnal hypoxia parameters. After overnight nCPAP treatment, plasma SFRP5 levels of OSA patients did not change significantly (t=1.557, P = 0.148) compared to that of pretreatment.Conclusions In nondiabetic OSA patients, plasma SFRP5 is associated with the lipid profile. However,no correlation was observed between SFRP5 and FBG or sleep parameters. The SFRP5 level of OSA patients did not differ from that of non-OSA individuals in our study.展开更多
The methanol inducible alcohol oxidase 1 promoter and the Saccharomyces cerevisiae alpha-factor prepro secretion signal were used to drive expression and secretion of a mucin-type fusion protein by Pichia pastoris in ...The methanol inducible alcohol oxidase 1 promoter and the Saccharomyces cerevisiae alpha-factor prepro secretion signal were used to drive expression and secretion of a mucin-type fusion protein by Pichia pastoris in 1 L scale bioreactors. The aim of the study was to understand how varying expression rates influenced the secretion dynamics of the fusion protein in terms of intracellular- and extracellular concentrations. Endoplasmic reticulum (ER) folding stress was assessed by the relative expression of the unfolded protein response controlled KAR2 gene. Three predefined methanol feeding models were applied to control the fusion protein synthesis rate. To track the fusion protein synthesis in a non-invasive manner and to follow its intracellular distribution, its C-terminal was linked to the green fluorescent protein. Under all conditions the fusion protein was found to partially accumulate intracellularly, where the major fraction was an insoluble, fluorescent full-sized protein. The high degree of glycosylation of the insoluble fusion protein indicated a secretory bottle-neck in the Golgi-system. This result was consistent with low ER folding stress as quantified by the relative expression of the KAR2 gene. Reduction of recombinant protein synthesis rate, by using lower feed rates of methanol, enhanced extracellular concentrations from 8 to 18 mg·L–1 and reduced the rate of intracellular accumulation. This clearly demonstrates the importance of tuning the synthesis rate with secretory bottle-necks to maintain secretion.展开更多
目的检测不同临床阶段2型糖尿病(T2DM)患者胰腺再生蛋白(Reg)水平,分析血清Reg与T2DM及其慢性并发症的关系。方法选取2012年9月—2014年12月东南大学附属中大医院及南京6个社区医院(红山、玄武、莫愁、夫子庙、中华门和徐家巷)收治的门...目的检测不同临床阶段2型糖尿病(T2DM)患者胰腺再生蛋白(Reg)水平,分析血清Reg与T2DM及其慢性并发症的关系。方法选取2012年9月—2014年12月东南大学附属中大医院及南京6个社区医院(红山、玄武、莫愁、夫子庙、中华门和徐家巷)收治的门诊、住院患者以及健康体检者,根据诊断标准与排除标准共纳入950名参与者,将其分为5组:健康对照组(HC组,n=97)、高危人群组(HR组,n=209)、糖调节受损组(IGR组,n=292)、初发T2DM组(Onset组,n=188)和长程T2DM无并发症组(LD-无并发症组,n=94)、长程T2DM有并发症组(LD-有并发症组,n=70)。采用酶联免疫吸附法(ELISA)测定受试者清晨空腹状态下的血清Reg水平,分析Reg与T2DM病程及慢性并发症的关系。结果各组年龄、糖尿病家族史、吸烟史、收缩压比较,差异均无统计学意义(P>0.05);各组性别、体质指数(BMI)、舒张压、空腹血糖(FPG)、餐后2 h血糖(2 h PG)、糖化血红蛋白(Hb A1c)、Reg比较,差异均有统计学意义(P<0.05)。T2DM患者血清Reg水平与病程呈正相关(rs=0.284,P<0.01)。在所有受试者中血清Reg水平与Hb A1c、FPG、2 h PG呈正相关(rs=0.188、0.115、0.111,P<0.001)。Reg诊断T2DM的ROC曲线下面积为0.640〔95%CI(0.605,0.674)〕,最佳截点为25 ng/ml,其灵敏度为46%,特异度为76%,阳性似然比为1.92,阴性似然比为0.71。Reg诊断T2DM慢性并发症的ROC曲线下面积为0.754〔95%CI(0.694,0.813)〕,最佳截点为27 ng/ml,其灵敏度为65%,特异度为74%,阳性似然比为2.50,阴性似然比为0.47。结论检测血清Reg水平有助于预测T2DM及其慢性并发症,Reg将来可能成为T2DM及其进展的一种预测因子。展开更多
Objective：To investigate the feasibility of the combination of detecting hypermethylated secreted frizzled-related protein 1 （SFRP1） and secreted frizzled-related protein 2（SFRP2） in feces as a panel of biomarker...Objective：To investigate the feasibility of the combination of detecting hypermethylated secreted frizzled-related protein 1 （SFRP1） and secreted frizzled-related protein 2（SFRP2） in feces as a panel of biomarkers for colorectal cancer（CRC） screening. Methods： Methylation-specific PCR（MSP） was performed to analyze methylation status of SFRP1 and SFRP2 in a blinded fashion in tumor tissues and in matched stool samples from 39 patients with primary CRC, 34 patients with adenomas, 17 patients with hyperplastic polyps and 20 endoscopically normal subjects as normal controls. Simultaneously we analyzed the correlation of hypermethylated SFRP1 and SFRP2 with the clinicopathological features of CRC. Results：Hypermethylated SFRP1 was detected in 92.3%, 76.5%, 47.1% of tissue samples and in 89.7%, 64.7%, 35.3% of matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. Hypermethylated SFRP2 was detected in 87.2%, 67.6%, 35.3% of tissue samples and in 82.1%, 55.9%, 29.4% of matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. Of these two genes, at least one hypermethylated was 94.9%, 82.4%, 52.9% in tissue samples and 92.3%,73.5%, 47.1% in matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. In contrast, no hypermethylated SFRP1 and SFRP2 were detected in mucosa tissues of normal controls, only 2 cases of fecal samples was detected with hypermethylated SFRP1 and another 1 case was detected with hypermethylated SFRP2. Moreover, no significant associations were observed between hypermethylated SFRP1,SFRP2 and clinicopathological features of CRC. Conclusion： Hypermethylation of SFRP1 and SFRP2 in feces are novel epigenetic biomarkers of CRC and carded high potential for the remote detection of CRC as non-invasive screening method, and combined analysis of hypermethylated SFRP1 and SFRP2 in fecal could further increase the detection rate of CRC and premalignant lesions.展开更多
目的探讨恩格列净联合利拉鲁肽对肥胖型2型糖尿病(type 2 diabetes,T2DM)患者糖脂代谢及血清分泌型卷曲相关蛋白5(secreted frizzled-related protein 5,SFRP5)、鸢尾素(Irisin)、总脂联素(total adiponectin,tAPN)水平的调节效应。方...目的探讨恩格列净联合利拉鲁肽对肥胖型2型糖尿病(type 2 diabetes,T2DM)患者糖脂代谢及血清分泌型卷曲相关蛋白5(secreted frizzled-related protein 5,SFRP5)、鸢尾素(Irisin)、总脂联素(total adiponectin,tAPN)水平的调节效应。方法选取2019年3月至2021年3月邯郸市中心医院肥胖型T2DM患者116例,以随机抽签法分为观察组(n=58)、对照组(n=58)。对照组给予利拉鲁肽治疗,观察组在对照组基础上联合恩格列净治疗,均治疗3个月。比较两组治疗前、治疗3个月后减重有关指标[体重、腰围、体重指数(body mass index,BMI)]、糖代谢指标[糖化血红蛋白(glycated hemoglobin,HbA1c)、空腹血糖(fasting blood glucose,FPG)、餐后2 h血糖(2 h postprandial blood glucose,2 hPG)]、脂代谢指标[低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、三酰甘油(triglycerides,TG)、总胆固醇(total cholesterol,TC)]、胰岛素敏感性有关指标[稳态模型胰岛β细胞功能指数(homeostasis model isletβcell function index,HOMA-β)、稳态模型胰岛素抵抗指数(homeostasis model insulin resistance index,HOMA-IR)]、血清SFRP5、Irisin、tAPN水平,并统计不良反应发生情况。结果观察组与对照组相比治疗3个月后体重[(74.93±2.68)kg比(78.21±3.47)kg]、腰围[(90.40±2.17)cm比(93.28±2.53)cm]、BMI[(26.57±0.94)kg/m2比(27.94±1.06)kg/m^(2)]、HbA1c[(6.03±0.59)%比(6.76±0.68)%]、FPG[(6.24±0.73)mmol/L比(6.97±0.95)mmol/L]、2 hPG[(8.06±1.25)mmol/L比(8.83±1.50)mmol/L]、LDL-C[(2.27±0.21)mmol/L比(2.58±0.27)mmol/L]、TG[(1.83±0.34)mmol/L比(2.20±0.48)mmol/L]、TC[(5.36±0.42)mmol/L比(5.92±0.53)mmol/L]、HOMA-IR(2.35±0.24比2.69±0.31)水平均下降(P<0.05),HDL-C[(1.50±0.38)mmol/L比(1.19±0.25)mmol/L]、HOMA-β(68.94±13.26比57.52±10.39)及血清SFRP5、Irisin、tAPN水平均升高(P<0.05);观察组治疗期间不良反应发生率17.24%与对照组12.07%相比,差异无统计学意义(P>0.05)。结论应用恩格列净联合利拉鲁肽治疗肥胖型T2DM患者可改善糖脂代谢与胰岛素敏感性,提高血清SFRP5、Irisin、tAPN水平,提升减重效果,且具有安全性。展开更多
基金Supported by National Natural Science Foundation of China,No. 81050016Research Fund for the Doctoral Program of Higher Education of China,No. 200800250003
文摘AIM： To investigate the expression and methylation status of the secreted frizzled-related protein 2 （SFRP2） in esophageal squamous cell carcinoma （ESCC） and ex- plore its role in ESCC carcinogenesis.METHODS： Seven ESCC cell lines （KYSE 30, KYSE150, KYSE410, KYSE510, EC109, EC9706 and TE-1） and one immortalized human esophageal epithelial cell line （Het- 1A）, 20 ESCC tissue samples and 20 paired adjacent non-tumor esophageal epithelial tissues were analyzed in this study. Reverse-transcription polymerase chain reaction （RT-PCR） was employed to investigate the expression of SFRP2 in cell lines, primary ESCC tumor tissue, and paired adjacent normal tissue. Methylation status was evaluated by methylation-specific PCR and bisulfite sequencing. The correlation between expres- sion and promoter methylation of the SFRP2 gene was confirmed with treatment of 5-aza-2＇-deoxycytidine. To assess the potential role of SFRP2 in ESCC, we es-tablished stable SFRP2-transfected cells and examined them with regard to cell proliferation, colony formation, apoptosis and cell cycle in vivo and in vitro.RESULTS： SFRP2 mRNA was expressed in the im- mortalized normal esophageal epithelial cell line but not in seven ESCC cell lines. By methylation-specific PCR, complete methylation was detected in three cell lines with silenced SFRP2 expression, and extensive methylation was observed in the other four ESCC cell lines. 5-aza-2＇-deoxycytidine could restore the expres- sion of SFRP2 mRNA in the three ESCC cell lines lack- ing SFRP2 expression. SFRP2 mRNA expression was obviously lower in primary ESCC tissue than in adjacent normal tissue （0.939 ± 0.398 vs 1.51 ± 0.399, P 〈 0.01）. SFRP2 methylation was higher in tumor tissue than in paired normal tissue （95% vs 65%, P 〈 0.05）. The DNA methylation status of the SFRP2 correlated inversely with the SFRP2 expression. To assess the potential role of SFRP2 in ESCC, we established stable SFRP2 transfectants and control counterparts by in- troducing pcDNA3.1/v5 hisA -SFRP2 or pcDNA3.1/v5 hisA -empty vector into KYSE30 cells lacking SFRP2 expression. After transfection, the forced-expression of SFRP2 was confirmed by the RT-PCR. In comparison with the control groups, stably-expressed SFRP2 in KYSE 30 cells significantly reduced colony formation in vitro （47.17% 4± 15.61% vs 17% ：1： 3.6%, P = 0.031） and tumor growth in nude mice （917.86：1：249.35 mm3 vs 337.23 ± 124.43 mm3, P 〈 0.05）. Using flow cytom- etry analysis, we found a significantly higher number of early apoptotic cells in SFRP2-transfected cells than in the control cells （P = 0.025）. The mean cell number in the S and G2-M phases of the cell cycle was also significantly lower in SFRP2-transfected KYSE30 cells compared with mock transfected counterparts. CONCLUSION： Silencing of SFRP2 expression through promoter hypermethylation may be a factor in ESCC carcinogenesis through loss of its tumor-suppressive activity.
基金supported by the Natural Science Research Project of colleges and Universities in Anhui Province[2022AH052336]High Level Talent Research Initiation Fund Of Anhui Medical College[2023RC004]。
文摘Objective Microcystin-leucine-arginine(MC-LR)exposure induces lipid metabolism disorders in the liver.Secreted frizzled-related protein 5(SFRP5)is a natural antagonist of winglesstype MMTV integration site family,member 5A(Wnt5a)and an anti-inflammatory adipocytokine.In this study,we aimed to investigate whether MC-LR can induce lipid metabolism disorders in hepatocytes and whether SFRP5,which has anti-inflammatory effects,can alleviate the effects of hepatic lipid metabolism by inhibiting the Wnt5a/Jun N-terminal kinase(JNK)pathway.Methods We exposed mice to MC-LR in vivo to induce liver lipid metabolism disorders.Subsequently,mouse hepatocytes that overexpressed SFRP5 or did not express SFRP5 were exposed to MC-LR,and the effects of SFRP5 overexpression on inflammation and Wnt5a/JNK activation by MC-LR were observed.Results MC-LR exposure induced liver lipid metabolism disorders in mice and significantly decreased SFRP5 mRNA and protein levels in a concentration-dependent manner.SFRP5 overexpression in AML12cells suppressed MC-LR-induced inflammation.Overexpression of SFRP5 also inhibited Wnt5a and phosphorylation of JNK.Conclusion MC-LR can induce lipid metabolism disorders in mice,and SFRP5 can attenuate lipid metabolism disorders in the mouse liver by inhibiting Wnt5a/JNK signaling.
基金Supported by the Special-purpose Scientific Research Foundation for University Doctorate Project of the Ministry of Education of China, No. 301090255
文摘AIM： To investigate the functions of promoter hypermethylation of secreted frizzled-related proteins （sFRPs） genes in colorectal tumorigenesis and progression. METHODS： The promoter hypermethylation and expression of sFRP genes in 72 sporadic colorectal carcinomas, 33 adenomas, 18 aberrant crypt foci （ACF） and colorectal cancer cell lines RKO, HCT116 and SW480 were detected by methylation-specific PCR and reverse transcription PCR, respectively. RESULTS： None of the normal colorectal mucosa tissues showed methylated bands of any of four sFRP genes, sFRP1, 2, 4 and 5 were frequently methylated in colorectal carcinoma, adenoma and ACF （sFRP1 〉 85%, sFRP2 〉75%, sFRP5 〉 50%）, and the differences between three colorectal tissues were not significant （P 〉 0.05）. IVlethylation in colorectal tumors was more frequent than in normal mucosa and adjacent normal mucosa. The mRNA of sFRP1-5 genes was expressed in all normal colorectal mucosa samples. Expression of sFRP1, 2, 4 and 5 and sFRP1, 2 and 5 was downregulated in carcinoma and adenoma, respectively. The downregulation of sFRP2, 4 and 5 was more frequent in carcinoma than in adenoma. Expression of sFRP3 which promoter has no CpG island was downregulated in only a few of colorectal tumor samples （7/105）. The downregulation ofsFRP1, 2, 4 and 5 expression was significantly associated with promoter hypermethylation in colorectal tumor. After cells were treated by DAC/TSA combination, the silenced sFRP mRNA expression could be effectively re-expressed in colorectal cancer cell lines. CONCLUSION： Hypermethylation of sFRP genes is a common early event in the evolution of colorectal tumor, occurring frequently in ACF, which is regarded as the earliest lesion of multistage colorectal carcinogenesis. It appears to functionally silence sFRP genes expression. Methylation of sFRP1, 2 and 5 genes might serve as indicators for colorectal tumor.
基金Supported by Liaoning Education Divison Foundation, No.05L557
文摘AIM： To identify the methylation of secreted frizzled-related protein 1 （SFRP1） in gastric cancer and to investigate the aberrant expression of SFRP1 and its correlation with the clinical pathological features of patients. METHODS： We determined SFRP1 methylation and SFRP1 mRNA expression in 3 gastric cancer cell lines SGC-7901, BGC-823, HGC-27, from 52 primary gastric cancer specimens and matched tumor adjacent tissue specimens by methylation-specific （MSP） PCR and RT-PCR respectively. Fisher＇s exact test was used to analyze the statistical association between clinical pathological data and aberrant expression of SFRP1. RESULTS： In 3 cancer cell lines, BGC-823 and HGC-27 had methylated SFRP1 and lost SFRP1 mRNA expression. After treatment of BGC-823 and HGC-27 with the demethylating agent, 5-aza-2′-deoxycytidine, SFRP1 was re-expressed. In 52 primary gastric cancer specimens and matched tumor adjacent tissue specimens, hypermethylation of SFRP1 was detected in 23 （44%） and 8 （15%） specimens respectively （x^2= 10.34, P 〈 0.01）. Loss of SFRP1 expression was detected in 17（33%） and 6 （12%） specimens respectively （x^2= 6.75, P 〈 0.01）. There was a significant correlation between SFRP1 hypermethylation and SFRP1 expression loss. SFRP1 expression was also correlated significantly with tumor stage and lymph node status, but not with patient sex, age and histological type. CONCLUSION： SFRP1 inactivation is a common and early event caused mainly by hypermethylation in gastric cancer. SFRP1 expression loss may be correlated with tumor metastasis in primary gastric cancer.
文摘Objective Obstructive sleep apnea (OSA) is closely related to obesity, insulin resistance and inflammation. Secreted frizzled-related protein 5 (SFRP5) is a recently discovered adipokine. It is involved in insulin resistance and inflammation in obesity. This study aimed at evaluating the association between SFRP5and sleeping characteristics as well as biochemical parameters of OSA patients.Methods This was a prospective case control study. Nondiabetic OSA patients and controls were consecutively recruited and divided into three groups: OSA group, apnea–hypopnea Index (AHI)≥5/h; healthy controls with normal body mass index (BMI); obese controls without OSA, and BMI > 24.0 kg/m2. All participants underwent polysomnography (PSG). Plasma SFRP5 was examined using enzyme-linked immunosorbent assay (ELISA). Blood biochemical examinations, including fasting blood glucose (FBG), lipid profile, hypersensitive Creactive protein (hsCRP), were performed early in the morning after PSG. Patients with severe OSA were treated with nasal continuous positive airway pressure (nCPAP), and plasma SFRP5 was repeatedly measured for comparison.Results Sixty-eight subjects were enrolled in the study, including 38 patients of OSA, whose medium AHI was 58.70 /h (36.63, 71.15), 20 obese controls, and 10 healthy controls. The plasma SFRP5 level of OSA patients was not significantly different from that of healthy controls or obese controls. In OSA patients, SFRP5 level correlated positively with triglyceride level (r=0.447, P=0.005) and negatively with LDL-cholesterol level and HDLcholesterol level (r=?0.472 and P=0.003; r=?0.478 and P=0.002; respectively). SFRP5 level was not found correlating with FBG, AHI, or any of nocturnal hypoxia parameters. After overnight nCPAP treatment, plasma SFRP5 levels of OSA patients did not change significantly (t=1.557, P = 0.148) compared to that of pretreatment.Conclusions In nondiabetic OSA patients, plasma SFRP5 is associated with the lipid profile. However,no correlation was observed between SFRP5 and FBG or sleep parameters. The SFRP5 level of OSA patients did not differ from that of non-OSA individuals in our study.
基金This work was supported by the Research Council of NorrbottenIn-novationsbron and Langmanska foretagarfonden.J.H.was supported by the Swedish Research Council(No.K2011-65X-3031-01-6)the County Council of Vastra Gotaland(ALF).
文摘The methanol inducible alcohol oxidase 1 promoter and the Saccharomyces cerevisiae alpha-factor prepro secretion signal were used to drive expression and secretion of a mucin-type fusion protein by Pichia pastoris in 1 L scale bioreactors. The aim of the study was to understand how varying expression rates influenced the secretion dynamics of the fusion protein in terms of intracellular- and extracellular concentrations. Endoplasmic reticulum (ER) folding stress was assessed by the relative expression of the unfolded protein response controlled KAR2 gene. Three predefined methanol feeding models were applied to control the fusion protein synthesis rate. To track the fusion protein synthesis in a non-invasive manner and to follow its intracellular distribution, its C-terminal was linked to the green fluorescent protein. Under all conditions the fusion protein was found to partially accumulate intracellularly, where the major fraction was an insoluble, fluorescent full-sized protein. The high degree of glycosylation of the insoluble fusion protein indicated a secretory bottle-neck in the Golgi-system. This result was consistent with low ER folding stress as quantified by the relative expression of the KAR2 gene. Reduction of recombinant protein synthesis rate, by using lower feed rates of methanol, enhanced extracellular concentrations from 8 to 18 mg·L–1 and reduced the rate of intracellular accumulation. This clearly demonstrates the importance of tuning the synthesis rate with secretory bottle-necks to maintain secretion.
文摘目的检测不同临床阶段2型糖尿病(T2DM)患者胰腺再生蛋白(Reg)水平,分析血清Reg与T2DM及其慢性并发症的关系。方法选取2012年9月—2014年12月东南大学附属中大医院及南京6个社区医院(红山、玄武、莫愁、夫子庙、中华门和徐家巷)收治的门诊、住院患者以及健康体检者,根据诊断标准与排除标准共纳入950名参与者,将其分为5组:健康对照组(HC组,n=97)、高危人群组(HR组,n=209)、糖调节受损组(IGR组,n=292)、初发T2DM组(Onset组,n=188)和长程T2DM无并发症组(LD-无并发症组,n=94)、长程T2DM有并发症组(LD-有并发症组,n=70)。采用酶联免疫吸附法(ELISA)测定受试者清晨空腹状态下的血清Reg水平,分析Reg与T2DM病程及慢性并发症的关系。结果各组年龄、糖尿病家族史、吸烟史、收缩压比较,差异均无统计学意义(P>0.05);各组性别、体质指数(BMI)、舒张压、空腹血糖(FPG)、餐后2 h血糖(2 h PG)、糖化血红蛋白(Hb A1c)、Reg比较,差异均有统计学意义(P<0.05)。T2DM患者血清Reg水平与病程呈正相关(rs=0.284,P<0.01)。在所有受试者中血清Reg水平与Hb A1c、FPG、2 h PG呈正相关(rs=0.188、0.115、0.111,P<0.001)。Reg诊断T2DM的ROC曲线下面积为0.640〔95%CI(0.605,0.674)〕,最佳截点为25 ng/ml,其灵敏度为46%,特异度为76%,阳性似然比为1.92,阴性似然比为0.71。Reg诊断T2DM慢性并发症的ROC曲线下面积为0.754〔95%CI(0.694,0.813)〕,最佳截点为27 ng/ml,其灵敏度为65%,特异度为74%,阳性似然比为2.50,阴性似然比为0.47。结论检测血清Reg水平有助于预测T2DM及其慢性并发症,Reg将来可能成为T2DM及其进展的一种预测因子。
基金supported by the grant from Programs of Science and Technology Commission Foundation of Jiangsu Province(NO.BS2005036)
文摘Objective：To investigate the feasibility of the combination of detecting hypermethylated secreted frizzled-related protein 1 （SFRP1） and secreted frizzled-related protein 2（SFRP2） in feces as a panel of biomarkers for colorectal cancer（CRC） screening. Methods： Methylation-specific PCR（MSP） was performed to analyze methylation status of SFRP1 and SFRP2 in a blinded fashion in tumor tissues and in matched stool samples from 39 patients with primary CRC, 34 patients with adenomas, 17 patients with hyperplastic polyps and 20 endoscopically normal subjects as normal controls. Simultaneously we analyzed the correlation of hypermethylated SFRP1 and SFRP2 with the clinicopathological features of CRC. Results：Hypermethylated SFRP1 was detected in 92.3%, 76.5%, 47.1% of tissue samples and in 89.7%, 64.7%, 35.3% of matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. Hypermethylated SFRP2 was detected in 87.2%, 67.6%, 35.3% of tissue samples and in 82.1%, 55.9%, 29.4% of matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. Of these two genes, at least one hypermethylated was 94.9%, 82.4%, 52.9% in tissue samples and 92.3%,73.5%, 47.1% in matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. In contrast, no hypermethylated SFRP1 and SFRP2 were detected in mucosa tissues of normal controls, only 2 cases of fecal samples was detected with hypermethylated SFRP1 and another 1 case was detected with hypermethylated SFRP2. Moreover, no significant associations were observed between hypermethylated SFRP1,SFRP2 and clinicopathological features of CRC. Conclusion： Hypermethylation of SFRP1 and SFRP2 in feces are novel epigenetic biomarkers of CRC and carded high potential for the remote detection of CRC as non-invasive screening method, and combined analysis of hypermethylated SFRP1 and SFRP2 in fecal could further increase the detection rate of CRC and premalignant lesions.
文摘目的探讨恩格列净联合利拉鲁肽对肥胖型2型糖尿病(type 2 diabetes,T2DM)患者糖脂代谢及血清分泌型卷曲相关蛋白5(secreted frizzled-related protein 5,SFRP5)、鸢尾素(Irisin)、总脂联素(total adiponectin,tAPN)水平的调节效应。方法选取2019年3月至2021年3月邯郸市中心医院肥胖型T2DM患者116例,以随机抽签法分为观察组(n=58)、对照组(n=58)。对照组给予利拉鲁肽治疗,观察组在对照组基础上联合恩格列净治疗,均治疗3个月。比较两组治疗前、治疗3个月后减重有关指标[体重、腰围、体重指数(body mass index,BMI)]、糖代谢指标[糖化血红蛋白(glycated hemoglobin,HbA1c)、空腹血糖(fasting blood glucose,FPG)、餐后2 h血糖(2 h postprandial blood glucose,2 hPG)]、脂代谢指标[低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、三酰甘油(triglycerides,TG)、总胆固醇(total cholesterol,TC)]、胰岛素敏感性有关指标[稳态模型胰岛β细胞功能指数(homeostasis model isletβcell function index,HOMA-β)、稳态模型胰岛素抵抗指数(homeostasis model insulin resistance index,HOMA-IR)]、血清SFRP5、Irisin、tAPN水平,并统计不良反应发生情况。结果观察组与对照组相比治疗3个月后体重[(74.93±2.68)kg比(78.21±3.47)kg]、腰围[(90.40±2.17)cm比(93.28±2.53)cm]、BMI[(26.57±0.94)kg/m2比(27.94±1.06)kg/m^(2)]、HbA1c[(6.03±0.59)%比(6.76±0.68)%]、FPG[(6.24±0.73)mmol/L比(6.97±0.95)mmol/L]、2 hPG[(8.06±1.25)mmol/L比(8.83±1.50)mmol/L]、LDL-C[(2.27±0.21)mmol/L比(2.58±0.27)mmol/L]、TG[(1.83±0.34)mmol/L比(2.20±0.48)mmol/L]、TC[(5.36±0.42)mmol/L比(5.92±0.53)mmol/L]、HOMA-IR(2.35±0.24比2.69±0.31)水平均下降(P<0.05),HDL-C[(1.50±0.38)mmol/L比(1.19±0.25)mmol/L]、HOMA-β(68.94±13.26比57.52±10.39)及血清SFRP5、Irisin、tAPN水平均升高(P<0.05);观察组治疗期间不良反应发生率17.24%与对照组12.07%相比,差异无统计学意义(P>0.05)。结论应用恩格列净联合利拉鲁肽治疗肥胖型T2DM患者可改善糖脂代谢与胰岛素敏感性,提高血清SFRP5、Irisin、tAPN水平,提升减重效果,且具有安全性。
