人粪便SFRP2基因甲基化分析对结直肠癌的诊断价值

目的探讨人粪便中分泌型卷曲相关蛋白2(SFRP2)基因甲基化分析用于结直肠癌(CRC)早期诊断的可行性。方法从87例结直肠癌或良性病变的患者及24例正常对照者的粪便中分别提取DNA,采用甲基化特异性PCR(MSP)技术分析其SFRP2基因甲基化状态。结果CRC、腺瘤、增生性息肉和溃疡性结肠炎患者的SFRP2基因甲基化阳性率分别为94.2%(49/52)、52.4%(11/21)、37.5%(3/8)和16.7%(1/6)。1例正常对照SFRP2基因甲基化检测阳性。检测SFRP2基因甲基化诊断CRC及癌前病变的敏感性和特异性分别为90.5%和85.4%。结论SFRP2基因甲基化是CRC进展过程中的早期事件。粪便SFRP2基因甲基化分析可望成为CRC早期无创诊断或CRC高风险人群筛查的新途径。     

粪便中SFRP2基因甲基化与大肠癌的相关性

目的探讨粪便中的分泌型卷曲相关蛋白(SFRP)2基因甲基化对大肠癌患者的诊断价值。方法选取2011年12月至2013年12月该院经病理检查确诊的52例大肠癌病人及32例结肠镜阴性的健康者(对照组)。通过亚硫酸氢盐的限制酶法(COBRA)检测大肠癌52例粪便中SFRP2基因启动子两个区域(区域1和区域2)甲基化状态,分析SFRP2基因甲基化与肿瘤临床病理因素的关系。结果大肠癌患者及健康者粪便中广泛甲基化、部分甲基化及无甲基化分别为59.6%(31/52)、84.6%(44/52)、15.4%(8/52)和12.5%(4/32),25.0%(8/32)、75.0%(24/32);广泛和部分SFRP2的甲基化率在大肠癌患者粪便的检出率均明显高于健康体检者(P0.05);SFRP2基因甲基化与大肠癌患者性别、肿瘤分期、浸润程度无显著关联。结论 SFRP2基因甲基化是大肠癌进展过程中的早期事件,粪便SFRP2基因甲基化有望成为早期无创诊断大肠癌的新途径。     

粪便SDC2、TFPI2、SFRP2基因甲基化联合检测在结直肠癌早期筛查中的临床价值

目的 探讨粪便SDC2、TFPI2、SFRP2基因甲基化联合检测在结直肠癌早期筛查中的临床价值。方法 收集苏州大学附属第一医院2021年6月至2022年2月就诊患者105例，其中结直肠癌组患者30例，腺瘤组患者45例，健康对照者30名，采用荧光定量PCR法检测患者粪便SDC2、TFPI2、SFRP2的甲基化情况。分析粪便SDC2、TFPI2、SFRP2甲基化及其三者联合检测诊断结直肠癌的敏感度及特异度，以病理结果为金标准，通过ROC曲线分析该检测方法对疾病的诊断效能。结果 结直肠癌组患者粪便SDC2、TFPI2、SFRP2甲基化及其联合结直肠癌检测阳性率高于腺瘤组和正常对照组，且腺瘤组高于正常对照组。SDC2、TFPI2、SFRP2甲基化及其联合检测结直肠癌的敏感度分别为46.7%、53.3%、66.7%、76.7%;特异度分别为96.7%、93.3%、96.7%、86.7%。ROC曲线分析显示粪便SDC2、SFRP2、TFPI2甲基化联合检测筛查结直肠癌的效能较高。结论 粪便SDC2、TFPI2、SFRP2甲基化联合检测对结直肠癌早期筛查的诊断价值较高，具有重要意义。     

联合检测vimentin和SFRP2甲基化在大肠癌筛查中的应用评价

目的评价在粪便样本中联合检测vimentin和SFRP2甲基化来筛查大肠癌的性能。方法搜集合格的新鲜粪便标本95例,其中40例大肠癌患者、25例进展期腺瘤患者和30例结肠镜阴性的正常人,应用甲基化特异性PCR(MSP)法检测vimentin和SFRP2甲基化状态,并与单个基因甲基化检测和粪隐血试验(FOBT)的诊断性能相比较是否有统计学差异。结果大肠癌组中vimentin和SFRP2甲基化阳性检出率分别为55.0%(22/40)和70.0%(28/40);进展期腺瘤组中为48.0%(12/25)和64.0%(16/25);正常对照组中为6.7%(2/30)和0(0/30)。在病例组中两基因联合检测的敏感性为83.1%(54/65),明显高于vimentin的52.3%(34/65)和SFRP2的67.7%(44/65),更要高于FOBT的27.7%(18/65)(均P0.05)。而在正常对照组中两基因联合检测的特异性为93.3%(28/30),与vimentin的93.3%(28/30)和SFRP2的100%(30/30)以及FOBT的96.7%(29/30)相比均无明显差异(均P0.05)。结论在大肠癌筛查中,联合检测粪便中vimentin和SFRP2基因甲基化状态明显优于单个基因和粪便潜血试验,具有潜在的应用价值。     

人粪便中波形蛋白基因甲基化在结直肠癌早期诊断中的意义

目的 分析结直肠癌患者粪便中波形蛋白基因甲基化状态,探讨其作为结直肠癌早期诊断分子标志物的可行性和临床意义.方法 从郑州大学第一附属医院收治的60例结直肠癌患者、门诊行结肠镜检查的17例结直肠腺瘤患者及30名正常对照者的粪便中分别提取DNA,采用甲基化特异性PCR(MSP)方法分析其波形蛋白基因甲基化状态.结果 60例结直肠癌患者粪便中波形蛋白基因甲基化阳性率为51.67％(31/60),17例结直肠腺瘤患者粪便中波形蛋白基因甲基化阳性率为4/17,30名正常对照中无一例检测到波形蛋白基因甲基化.结论 粪便中波形蛋白基因甲基化是结直肠癌进展过程中的早期事件,粪便中波形蛋白基因甲基化分析有望成为结直肠癌早期诊断的标志物之一.     

人粪便中OSMR和TFPI2基因甲基化在结直肠癌诊断中的意义

目的:探讨检测人粪便中抑瘤素M型受体(OSMR)基因和组织因子途径抑制子2(TFPI2)基因甲基化对于结直肠癌及腺瘤诊断的可行性及临床意义.方法:从60例结直肠癌患者和17例腺瘤患者及30名正常对照者的粪便中分别提取DNA,应用甲基化特异性PCR方法检测人粪便中OSMR和TFPI2基因的甲基化状态.结果:结直肠癌患者粪便中OSMR及TFPI2基因甲基化检出率分别高于腺瘤患者和正常对照者[OSMR:35%(21/60)vs12%(2/17),7%(2/30);TFPI2:70%(42/60)vs18%(3/17),3%(1/30);均P<0.01].二者联合检测甲基化检出率为81.7%(49/60),特异性90%.结论:检测粪便中OSMR和TFPI2基因甲基化在结直肠癌诊断和筛查中有潜在的应用价值.     

粪便标本中 SFRP1和 WIF-1基因启动子甲基化在大肠癌早期筛查中的意义

目的：探讨联合检测粪便中分泌型卷曲蛋白1（SFRP1）和Wnt途径抑制因子-1（WIF-1）基因启动子甲基化作为大肠癌（ CRC）早期筛查指标的可行性。方法收集48例CRC患者、35例大肠腺瘤患者、32例大肠增生性息肉患者及30例健康人的粪便样本,采用甲基化特异性PCR技术分析样本中SFRP1和WIF-1基因启动子甲基化状态。结果 CRC患者粪便中SFRP1和WIF-1基因启动子甲基化率分别为68．8％和60．4％,腺瘤患者分别为57．1％和42．9％,增生性息肉患者分别为21．9％和18．8％;两者联合检测CRC和大肠腺瘤的敏感度分别为77．1％和65．7％,特异度为93．3％。2个基因甲基化水平在CRC和腺瘤患者的粪便样本中均显著高于增生性息肉和健康人群（P均＜0．05）。此外,SFRP1和WIF-1基因甲基化与临床病理特征无明显相关性。结论检测粪便中SFRP1和WIF-1基因启动子甲基化有望成为CRC早期筛查的一种非侵入性检测方法,在早期筛查和诊断中具有潜在的应用价值。     

粪便DNA中MGMT、XAF1基因启动子甲基化联合检测在结直肠癌筛查中的应用研究

目的使用甲基化特异性PCR(MSP)方法检测粪便DNA中O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)、X染色体连锁凋亡抑制蛋白相关因子1(XAF1)基因启动子甲基化情况,并探讨其在结直肠肿瘤诊断中的意义。方法收集40例结直肠腺癌、40例腺瘤性息肉、52例正常对照住院患者粪便标本,使用试剂盒提取其粪便中肠道脱离细胞DNA,通过MSP方法检测其MGMT、XAF1基因启动子甲基化情况。结果 MGMT、XAF1基因启动子甲基化在结直肠腺癌中的阳性率分别为50.0%、55.9%;在腺瘤性息肉中的阳性率分别为42.1%、52.6%;二者联合检测在结直肠腺癌及腺瘤性息肉中的阳性率分别为73.5%、68.4%;特异性为52.0%。结直肠腺癌患者粪便中粪便潜血阳性率为35.3%,CEA阳性率为35.3%。结论通过试剂盒提取粪便DNA具有较高成功率;粪便DNA中MGMT、XAF1基因启动子甲基化状态用于检测结直肠腺癌及腺瘤性息肉具有较高的敏感性。检测粪便基因甲基化有望成为CRC高风险人群筛查的一个重要途径。     

粪便Vimentin和SFRP2甲基化在结直肠癌筛查中的作用

目的探讨粪便中Vimentin和SFRP2甲基化状态在结直肠癌筛查中的价值。方法收集患者合格的粪便标本69例,其中结直肠癌23例、进展期腺瘤24例和健康人群22名,采用甲基化特异性PCR技术分析Vimentin和SFRP2甲基化状态,并与单个基因甲基化和粪便免疫隐血试验(FIT)的检测性能相比较,评价其在结直肠癌筛查中的灵敏度和特异度。结果结直肠癌组中单个Vimentin和SFRP2甲基化检出率分别为82.6%和69.6%,进展期腺瘤组为62.5%和41.7%,正常对照组为13.6%和13.6%。Vimentin和SFRP2联合检测在结直肠癌组的灵敏度为87.0%,高于FIT的56.5%(χ~2=5.25,P0.05),与单基因检测比较,差异无统计学意义(P0.05)。进展期腺瘤组中,联合检测的灵敏度为70.8%,高于SFRP2的41.7%(χ~2=4.15,P0.05)和FIT的29.2%(χ~2=8.33,P0.01),与Vimentin检测比较差异无统计学意义(P0.05)。正常对照组联合检测的特异度为86.4%,与单基因检测相同,与FIT(72.7%)比较,差异无统计学意义(P0.05)。联合检测在管状腺瘤中检出率为92.3%,高于SFRP2的53.8%(χ~2=4.9,P0.05),与Vimentin(76.9%)比较,差异无统计学意义(P0.05),在绒毛状管状腺瘤和管状腺瘤/绒毛状管状腺瘤中检出率与Vimentin相同,与SFRP2比较,差异无统计学意义(P0.05)。联合检测在伴有上皮内瘤变中的检出率与单基因检测差异均无统计学意义(P0.05)。结论粪便中联合Vimentin和SFRP2检测优于单基因及FIT检测,在结直肠癌筛查中具有潜在的应用价值。     

人粪便中UNC5C基因甲基化在早期诊断大肠癌中的临床意义

目的探讨人粪便中UNC5C基因甲基化在临床上早期诊断大肠癌的临床意义。方法选取2011年12月至2013年12月该院收治的大肠癌病人52例,健康体检者40例,对UNC5C启动子的两个区域(区域1和区域2),使用结合重亚硫酸盐限制性内切酶法(COBRA)研究对象的粪便标本中UNC5C基因甲基化状态。结果广泛UNC5C的甲基化及部分甲基化在大肠癌患者粪便、健康体检者中分为54%(28/52)和81%(42/52);0%(0/40)和10%(4/40)。无论广泛还是部分UNC5C的甲基化率大肠癌患者粪便的检出率均明显高于健康体检者(P<0.05)。人粪便UNC5C基因甲基化状态改变与大肠癌患者的性别、年龄、肿块大小没有明显的相关性(P>0.05)。结论无论广泛还是部分UNC5C的甲基化率大肠癌患者粪便的检出率均明显高于健康体检者,而广泛甲基化为大肠癌所特有。     

Detection of aberrant methylation in fecal DNA as a molecular screening tool for colorectal cancer and precancerous lesions

AIM: To investigate the feasibility of detecting methylated fecal DNA as a screening tool for colorectal carcinoma (CRC) and precancerous lesions. METHODS: Methylated secreted frizzled-related protein gene 2 (SFRP2 ), hyperplastic polyposis protein gene (HPP1 ) and O6-methylguanine-DNA methyltransferase gene (MGMT) in stools from 52 patients with CRC, 35 patients with benign colorectal diseases and 24 normal individuals were analyzed using methylation-specific PCR. RESULTS: Methylated SFRP2, HPP1 and MGMT were detected in 94.2%, 71.2%, 48.1% of CRC patients and 52.4%, 57.1%, 28.6% of adenoma patients, respectively. The overall prevalence of fecal DNA with at least one methylated gene was 96.2% and 81.8% in patients with CRC and precancerous lesions, respectively. In contrast, only one of the 24 normal individuals revealed methylated DNA. These results indicated a 93.7% sensitivity and a 77.1% specificity of the assay for detecting CRC and precancerous lesions. CONCLUSION: Methylation testing of fecal DNA using a panel of epigenetic markers may be a simple and promising non-invasive screening method for CRC and precancerous lesions.     

Hypermethylated SFRP2 gene in fecal DNA is a high potential biomarker for colorectal cancer noninvasive screening

AIM： To investigate the feasibility of detecting hypermethylated secreted frizzled-related protein 2 （SFRP2） gene in fecal DNA as a non-invasive screening tool for colorectal cancer （CRC）. METHODS： Fluorescence-based real-time PCR assay （MethyLight） was performed to analyze SFRP2 gene promoter methylation status in a blinded fashion in tumor tissues and in stool samples taken from 69 CRC patients preoperatively and at the 9th postoperative day, 34 patients with adenoma ≥1 cm, 26 with hyperplastic polyp, and 30 endoscopically normal subjects. Simultaneously the relationship between hypermethylation of SFRP2 gene and clinicopathological features was analyzed. RESULTS： SFRP2 gene was hypermethylated in 91.3% （63/69） CRC, 79.4% （27/34） and 53.8% （14/26） adenoma and hyperplastic polyp tissues, and in 87.0% （60/69）, 61.8% （21/34） and 42.3% （11/26） of corresponding fecal samples, respectively. In contrast, no methylated SFRP2 gene was detected in mucosal tissues of normal controls, while two cases of matched fecal samples from normal controls were detected with hypermethylated SFRP2. A significant decrease （P 〈 0.001） in the rate of hypermethylated SFRP2 gene was detected in the postoperative （8.7%, 6/69） fecal samples as compared with the preoperative fecal samples （87%, 60/69） of CRC patients. Moreover, no significant associations were observed between SFRP2 hypermethylation and clinicopathological features including sex, age, tumor stage, site, lymph node status and histological grade, etc.CONCLUSION： Hypermethylation of SFRP2 gene in fecal DNA is a novel molecular biomarker of CRC and carries a high potential for the remote detection of CRC and premalignant lesions as noninvasive screening method.     

Detection of promoter hypermethylation of Wnt antagonist genes in fecal samples for diagnosis of early colorectal cancer

AIM: To investigate the feasibility of detecting aberrantly hypermethylated Wnt-antagonist gene promoters (SFRP2 and WIF-1) in fecal DNA as non-invasive biomarkers for early colorectal cancer (CRC).METHODS: The methylation-specific polymerase chain reaction assay was performed to blindly analyze the methylation status of SFRP2 and WIF-1 gene promoters in fecal samples from 48 subjects with CRC, 35 with adenomas, 32 with hyperplastic polyps and 30 endoscopically normal subjects. Additionally, we compared the diagnostic efficiency of measuring the hypermethylated SFRP2 and WIF-1 genes in the feces to the fecal occult blood test (FOBT) for the early detection of CRC.RESULTS: Hypermethylated SFRP2 was detected in the feces of 56.3% (27/48) of CRC cases, 51.4% (18/35) of adenoma cases and 12.5% (4/32) of patients with hyperplastic polyps. The hypermethylation of WIF-1 was detected in 60.4% (29/48), 45.7% (16/35) and 18.7% (6/32) of fecal samples from CRC, adenoma and hyperplastic polyp patients, respectively. At least one hypermethylated gene was detected in 81.3% (39/48) of CRC and 65.7% (23/35) of adenoma samples. In contrast, only a hypermethylated WIF-1 gene was detected in one case of normal fecal samples. Moreover, no significant associations were observed between SFPR2 and WIF-1 hypermethylation and clinicopathological features. Additionally, 81.8% of CRC cases diagnosed as Dukes A stage or advanced adenomas had at least one hypermethylated gene detected, while the detection rate with the FOBT was only 31.8% (P < 0.001).CONCLUSION: Hypermethylated SFRP2 and WIF-1 genes in fecal DNA are novel and promising molecular biomarkers that have great diagnostic potential for early CRC.     

SFRP2 methylation in fecal DNA—a marker for colorectal polyps

Introduction DNA methylation of secreted frizzled-related proteins (SFRPs) can be detected in colorectal cancer (CRC) tissue, in tissue of adenomas, and in aberrant crypt foci, whereas in normal colorectal mucosa tissue, SFRP genes are unmethylated. Recently, our study group was able to demonstrate SFRP2 methylation as the most sensitive single DNA-based marker in stool for identification of CRC. The purpose of this study was to clarify whether SFRP2 methylation in fecal DNA can be found in stool of individuals with hyperplastic and adenomatous colorectal polyps. Materials and methods Patients who were diagnosed with colorectal polyps or showed negative colonoscopy were included in this study. DNA from stool samples was isolated. SFRP2 methylation was assessed by means of MethyLight. Results Stool samples from 68 individuals were checked for DNA content; 23% of the samples (6 of 26) from healthy controls, 46% of the samples (6 of 13) from patients with hyperplastic polyps, and 45% of the samples (13 of 29) from patients with adenomas were positive for human DNA. SFRP2 methylation in stool samples was found in none of the healthy controls, in 33% (2 of 6) patients with hyperplastic polyps, and in 46% (6 of 13) patients with adenomas. Statistical analysis revealed that the frequency of SFRP2 methylation increased significantly (P = 0.028) from healthy controls to patients with hyperplastic polyps and to patients with adenomas. Conclusions In the current study, we report for the first time that SFRP2 methylation in fecal DNA increases significantly from healthy controls to patients with hyperplastic polyps and to patients with adenomas. SFRP2 methylation may serve as a marker for molecular stool-based adenoma and CRC screening.     

Detection of hypermethylated DNA or cyclooxygenase-2 messenger RNA in fecal samples of patients with colorectal cancer or polyps

BACKGROUND: Detection of fecal DNA is a promising approach to colorectal cancer screening. However, the sensitivity of current fecal DNA tests for colorectal polyps is low. We evaluated the feasibility of detecting aberrantly methylated DNA or cyclooxygenase-2 (COX-2) mRNA in feces of patients with colorectal cancer or polyps. METHODS: Fecal samples were collected prior to colonoscopy from 20 patients with colorectal cancer, 30 patients with colorectal polyps, and 30 subjects with normal colonic examination. Presence of hypermethylated DNA in 7 tumor-related genes (APC, ATM, hMLH1, sFRP2, HLTF, MGMT, and GSTP1) in stool was analyzed by methylation-specific PCR. COX-2 mRNA in fecal samples was detected by RT-PCR. RESULTS: With the use of this panel of methylation markers, the sensitivity of detecting colorectal cancer and adenoma was 75% (95% CI 50.9-91.3%) and 68% (95% CI 46.5-85.1%), respectively. Three normal subjects also had methylated DNA detected in stool, which gives a specificity of 90% (95% CI 73.5-97.9%). The mean number of genes methylated in DNA from the stool of patients with colorectal cancer and adenoma was 1.4 and 0.9, respectively. In contrast, COX-2 mRNA was detected in the stool samples of 10 (50%) cancer patients and one (4%) patient with advanced adenoma only. Two (6.7%) stool samples from normal subjects also had COX-2 mRNA detected. CONCLUSION: Detection of aberrantly methylated DNA in fecal samples is more sensitive than COX-2 mRNA for detection of colorectal cancer and adenoma.     

Hypermethylation of <Emphasis Type="Italic">SFRP2</Emphasis> as a Potential Marker for Stool-Based Detection of Colorectal Cancer and Precancerous Lesions

DNA methylation is a key mechanism of colorectal carcinogenesis. Analysis of aberrantly methylation in stool DNA might provide a novel strategy for noninvasive detection of colorectal cancer (CRC). To explore the feasibility of this approach, we have assessed the methylation status of secreted frizzled-related protein gene 2 (SFRP2) in stool samples from patients with CRC with respect to a series of healthy individuals and patients with benign colorectal diseases, using methylation-specific polymerase chain reaction. Methylated SFRP2 occurs in 94.2%, 52.4%, 37.5%, and 16.7% of patients with CRC, adenomas, hyperplstic polyps, and ulcerative colitis, respectively. Of the 24 normal individuals, only 1 revealed methylated DNA. The pilot study revealed that aberrant methylated SFRP2 could be detected frequently in stools from patients with CRC and precancerous lesions. Methylation testing of fecal DNA may be a simple, promising, and noninvasive screening tool for colorectal neoplasia.