外周血SEPT9 DNA甲基化检测在结直肠癌筛查中的应用

结直肠癌是多种致癌因素作用下发生的恶性肿瘤,预后差,病死率较高。常用的结直肠癌筛查方法包括普通结肠镜检查、弯曲乙状结肠镜检查、愈创木脂化学法粪隐血试验、免疫化学粪隐血试验、粪便DNA检测、CT结肠成像以及血清癌胚抗原检测等,但这些检查均有缺陷。SEPT9 DNA甲基化水平升高与结直肠癌的发生相关,检测外周血SEPT9 DNA甲基化水平进行结直肠癌筛查,适合在易感人群中推广普及。本文就外周血SEPT9 DNA甲基化检测在结直肠癌筛查中的应用作一综述。     

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。     

SEPT9甲基化与多种血清肿瘤标志物对结直肠癌诊断价值的比较

[目的]探讨单一血浆SEPT9甲基化检测与多种血清学肿瘤标物在结直肠癌临床诊断中的意义,并对其诊断价值进行比较。[方法]选取82例不同分期的结直肠患者,64例良性息肉患者和60例正常对照,均行肠镜检查确认病理,采用定量甲基化特异性PCR检测SEPT9甲基化水平,采用化学发光法同时检测血清学中CEA、CA125、CA19-9、CA72-4、CA50和CA242六种血清学肿瘤标记物水平,并做统计学分析。[结果]血浆SEPT9甲基化检测对结直肠癌的检测灵敏度为:82.9%,单一的血清学肿瘤标物CEA、CA125、CA19-9、CA72-4、CA50和CA242对结直肠癌的检测灵敏度分别为35.4%、9.8%、18.3%、17.1%、13.4%和20.7%。6种血清学肿瘤标记物联合检测的灵敏度为57.3%。统计学分析显示,通过血清学6种肿瘤标记物联合检测后,其阳性检出率与任意一个单一肿瘤标记物的阳性检出率差异有统计学意义(P0.05)。与结直肠癌组比较,对照组6种血清学肿瘤标记物联合检测的阳性率差异有统计学意义(P0.05)。SEPT9甲基化检测对结直肠癌的检出率与6种单一的肿瘤标志物检出率差异有统计学意义(P0.05),且与6种肿瘤标记物的联合检测阳性率差异有统计学意义(P0.05)。与对照组比较,癌症组血浆SEPT9检测的阳性率差异有统计学意义(P0.01)。[结论]临床常用的单一肿瘤标物对结直肠癌的诊断意义不大,联合检测能提升诊断价值;而单独的血浆SEPT9甲基化检测对结直肠癌的诊断价值高于上述6种标记物联合检测,可以作为结直肠癌临床辅助诊断的一种理想手段。     

外周血SEPT9基因甲基化检测在结直肠癌筛查中的应用进展

结直肠癌(CRC)的早期诊断具有重要临床意义。外周血SEPT9基因甲基化在大量研究中被证实是CRC的特异性生物学标记物,敏感性和特异性均较高,较之目前常用的粪便隐血试验、血清肿瘤标记物和结肠镜筛查具有一定优势,但在筛查腺瘤、息肉等癌前病变中的应用价值有限,用于评估CRC术后复发和放化疗效果的潜力则尚需进一步临床验证。本文就外周血SEPT9基因甲基化检测在CRC筛查中的应用进展作一综述。     

伺机性筛查新模式在结直肠肿瘤患者一级亲属中的探索性研究

目的探究结直肠病房筛查新模式在结直肠肿瘤患者一级亲属筛查的有效性。 方法采用结直肠肿瘤风险问卷评分、粪便潜血免疫化学检测（FIT）以及粪便多靶点FIT-DNA检测对2019年10月至2021年7月在中国医学科学院肿瘤医院结直肠外科就诊的结直肠癌及进展期腺瘤患者的一级亲属进行检测，根据检测结果将一级亲属进行筛查风险分层以及肠镜检查推荐分类，分析不同分层分类后一级亲属的肠镜依从率与病变检出率。 结果共250名受试者被纳入本研究。总体人群肠镜依从率为38.0%（95/250），肠镜病变检出率为9.5%（9/95）；高风险人群（A类推荐人群）肠镜依从率为78.9%（15/19），肠镜病变检出率为26.7%（4/15）；中风险人群（B类推荐人群）肠镜依从率为61.2%（30/49），肠镜病变检出率为16.7%（5/30）；低风险人群（C类推荐人群）肠镜依从率为27.5%（50/182），肠镜病变检出率为0（0/50）。 结论三种筛查方法联合使用可以高效精准地区分一级亲属的筛查风险，此方案是一个可以在病房开展的有效可行的结直肠肿瘤患者一级亲属人群的伺机性筛查新模式。     

一氧化氮、一氧化氮合成酶与结直肠肿瘤

一氧化氮（NO）的生物学活性广泛，参与结直肠肿瘤的发生，发展，转移等过程，它能调控血管的生成，参与微循环的调节，在肿瘤免疫，细胞凋亡过程中发挥作用，对NO和一氧化氮合成酶抑制剂的研究，将为结直肠肿瘤的治疗提供新的思路。     

外周血SEPT9基因甲基化检测联合糖蛋白肿瘤标志物在结直肠癌诊断中的应用

背景:SEPT9基因甲基化是结直肠癌的特异性生物标志物,测定其外周血表达水平可评估受检者结直肠癌的患病风险。目的:初步探讨外周血SEPT9基因甲基化检测联合糖蛋白肿瘤标志物在结直肠癌诊断中的临床意义。方法:纳入2016年12月—2017年4月苏州大学附属第一医院诊断的正常对照组57例、腺瘤组61例和结直肠癌组71例患者,应用PCR荧光探针法测定外周血SEPT9基因甲基化情况,测定肿瘤标志物CEA、CA125、CA19-9、CA72-4、CA211水平,采用ROC曲线分析各指标诊断结直肠癌的价值。建立Logistic回归方程,评估SEPT9联合CEA诊断结直肠癌的价值。结果:正常对照组、腺瘤组、结直肠癌组的SEPT9基因甲基化阳性率分别为0、6. 6%、52. 1%。结直肠癌组CEA水平显著高于正常对照组、腺瘤组(P 0. 05)。ROC曲线分析显示,SEPT9、CEA以及两者联合诊断结直肠癌的AUC分别为0. 817、0. 707和0. 793。Logistic回归分析显示,SEPT9、CEA诊断结直肠癌的OR值分别为1. 394(1. 198～1. 762)、1. 325(0. 997～1. 622)。结论:随着疾病进展,外周血SEPT9基因甲基化率明显升高,对结直肠癌的诊断价值较高。联合外周血SEPT9基因甲基化和CEA有助于提高结直肠癌的早期筛查率。     

伺机性筛查新模式在结直肠肿瘤患者配偶的探索性研究

目的探究结直肠病房筛查新模式在结直肠肿瘤患者配偶筛查的有效性。 方法采用结直肠肿瘤风险问卷调查、粪便潜血免疫化学检测（FIT）以及粪便多靶点FIT-DNA检测对2019年10月至2021年7月在中国医学科学院肿瘤医院结直肠外科就诊的结直肠癌及进展期腺瘤患者的配偶进行检测,根据检测结果将配偶的筛查风险以及肠镜检查推荐分为A、B、C、D四类,分析不同分类后配偶肠镜依从率与病变检出率。 结果共206名受试者被纳入本研究。总体配偶人群肠镜依从率为29.6%（61/206）,肠镜病变检出率为9.8%（6/61）;A类至D类推荐人群肠镜依从率分别为90.9%（10/11）、53.5%（23/43）、20.5%（23/112）和12.5%（5/40）,肠镜病变检出率分别为30.0%（3/10）、8.7%（2/23）、4.3%（1/23）和0（0/5）。 结论三种筛查方法联合使用可以高效精准地区分配偶的筛查风险,此方案是一个可以在病房开展的有效可行的结直肠肿瘤患者配偶人群的伺机性筛查新模式。     

生物钟基因hClock蛋白在结直肠肿瘤中的表达研究

目的探讨人类生物钟基因hClock蛋白在结直肠肿瘤中的表达及其意义。方法应用免疫组织化学法检测不同Dukes分期结直肠肿瘤及相应癌旁组织中hClock基因蛋白产物（Clock蛋白）的表达，并比较其差异。结果Clock蛋白红结直肠肿瘤中呈现中或强阳性表达，与肿瘤分化程度及Dukes分期无显著相关性（P〉0.05）。癌旁组织呈现弱阳性表达。结论Clock蛋白与结直肠肿瘤的发生有相关性，与侵袭、转移的关系有待更深入地研究。     

一氧化氮、一氧化氮合成酶与结直肠肿瘤

一氧化氮(NO)的生物学活性广泛,参与结直肠肿瘤的发生、发展、转移等过程。它能调控血管的生成,参与微循环的调节,在肿瘤免疫、细胞凋亡过程中发挥作用。对NO和一氧化氮合成酶抑制剂的研究,将为结直肠肿瘤的治疗提供新的思路。     

几种粪便脱落细胞检查方法的大肠癌筛检效率比较

目的 评价不同粪便脱落细胞检查方法的大肠癌筛检效率.方法 对814例结肠镜检查者分别经清肠液法、二次自然粪便法、磁选法、自然排便联合清肠液液基法提取粪便脱落细胞,进行细胞学诊断.部分经清肠液法受检者提取细胞后行DNA核定量分析(清肠液+DNA核定量分析法).部分二次自然粪便法提取的粪便行基因检测.二次自然粪便法和自然排便联合清肠液液基法提取的粪便同时进行粪隐血检测.结果 粪便脱落细胞检出大肠癌的敏感性为66.27%(112/169),特异性为99.56%(225/226).大肠癌患者粪便中提取异犁细胞的阳件率与大肠癌分化程度和Duke分期无关(P＞0.05).二次自然排便法、自然排便联合清肠液液基法、清肠液+DNA核定量分析法筛检大肠癌的敏感性(分别为76.09%、68.29%、76.92%)优于单纯清肠液法和磁选法(分别为26.31%、43.24%).粪便脱落细胞p53、K-ras、腺瘤性息肉病(APC)基因检测阳性率与癌组织相应基因检测的一致率为83.33%(25/30)、9/10、9/10.基因检测的阳性率低于粪便脱落细胞学检测.联合进行粪隐血和粪脱落细胞学检测检出大肠癌的敏感性(93.10%)高于单独进行粪隐血试验(80.46%)和粪便脱落细胞学检测(73.56%).结论 粪便脱落细胞学检查是大肠癌筛检的有效手段.利用液基薄层技术对患者自然排便或清肠液进行两次筛查并联合粪隐血检测是最佳筛检方案.     

Systematic review of blood diagnostic markers in colorectal cancer

The purpose of this systematic review was to compare the diagnostic ability of blood markers for colorectal cancer (CRC). A systematic review of the literature for diagnostic blood markers for primary human colorectal cancer over the last 5 years was performed. The primary outcome was to assess the diagnostic ability of these markers in diagnosing colorectal cancer. The secondary outcome was to see whether the marker was compared to other markers. The tertiary outcome was to assess diagnostic ability in early versus late CRC, including stage IV disease. We identified 51 studies (29 prospective, 14 retrospective, and 8 meta-analyses). The markers were divided in broadly four groups: nucleic acids (RNA/DNA/messenger RNA/microRNAs), cytokines, antibodies, and proteins. The most promising circulating markers identified among the nucleid acids were NEAT_v2 non-coding RNA, SDC2 methylated DNA, and SEPT9 methylated DNA. The most promising cytokine to detect CRC was interleukin 8, and the most promising circulating proteins were CA11-19 glycoprotein and DC-SIGN/DC-SIGNR. Sensitivities of these markers for detecting primary colorectal carcinoma ranged from 70 to 98% and specificities from 84 to 98.7%. The best studied blood marker was SEPT9 methylated DNA, which showed great variability with sensitivities ranging from 48.2 to 95.6% and specificities from 80 to 98.9%, making its clinical applicability challenging. If combined with fecal immunochemical test (FIT), the sensitivity improved from 78 to 94% in detecting CRC. Methylated SEPT9, methylated SDC2, and -SIGN/DC-SIGNR protein had better sensitivity and specificity than CEA or CA 19-9. With the exception of SEPT9 which is currently being implemented as a screening test for CRC all other markers lacked reproducibility and standardization and were studied in relatively small population samples.     

Diagnostic value of occult fecal blood testing for colorectal cancer screening

AIM: To evaluate the diagnostic value of occult fecal blood testing in mass colorectal cancer screening.METHODS: A reverse passive hemagglutination reaction fecal occult blood test (RPHA-FOBT) and colorectal cancer risk factor quantitative method were used as preliminary screening for colorectal cancer. A 60-cm fiber optic colonoscopy was used to validate the preliminary screen and was used to detect colorectal cancer in a community of 75813 subjects.RESULTS: Compared to the 60-cm fiber optic colonoscopy as a standard reference, FOBT has a sensitivity of 41.9%, specificity of 95.8%, Youden′s index of 0.38, and positive predictive value of 0.68%. These results increased with subject age from the first detection. A 3-year follow up in the target mass showed that all new cases had initially been FOBT-negative.CONCLUSION: The value of FOBT as an indicator of colorectal cancer in mass screening is limited.     

Evaluation of Calgranulin B in Stools from the Patients with Colorectal Cancer

Purpose  Calprotectin (heterodimer of calgranulin A and B) has been previously studied as a candidate stool marker for detecting colorectal cancer. We assessed the clinical usefulness of calgranulin B as a stool marker for colorectal cancer in a pilot study of patients with colorectal cancer. Methods  We performed 2-DE-based proteomics to screen stool markers for colorectal cancer. We checked the calgranulin B in stools from 77 colorectal cancer patients and from 75 controls by western blot and enzyme-linked immunosorbent assay. We measured calgranulin A using the same methods, and stool hemoglobin by immunologic fecal occult blood test. Results  Fecal calgranulin A did not show any difference, but stool calgranulin B of colorectal cancer patients was significantly higher than controls [50.6 ng/mg stool protein (SD, 34.8) vs. 20.2 ng/mg stool protein (SD,24.0), respectively, P < 0.001). At the cut off level 24.4 ng/mg stool protein, the sensitivity was somewhat higher than fecal occult blood test (72.0 percent vs. 62.3 percent) but the specificity was much lower than fecal occult blood test (77.1 percent vs. 98.7 percent). Conclusions  Calgranulin B was increased in stools of colorectal cancer patients but our results suggest that colorectal cancer screening by determination of stool calgranulin B would not be better than conventional fecal occult blood test. This work was supported by research grants 0710670–1 (Yoo BC) and 0410063–3 (Lim SB) from the National Cancer Center, Korea. Poster presentation at the meeting of European Biomarkers Summit and Proteomics Europe, Amsterdam, The Netherlands, September 4 to 5, 2007.     

Measuring the Quality of Colorectal Cancer Screening: The Importance of Follow-Up

Purpose As evidence mounts for effectiveness, an increasing proportion of the United States population undergoes colorectal cancer screening. However, relatively little is known about rates of follow-up after abnormal results from initial screening tests. This study examines patterns of colorectal cancer screening and follow-up within the nation's largest integrated health care system: the Veterans Health Administration. Methods We obtained information about patients who received colorectal cancer screening in the Veterans Health Administration from an existing quality improvement program and from the Veterans Health Administration's electronic medical record. Linking these data, we analyzed receipt of screening and follow-up testing after a positive fecal occult blood test. Results A total of 39,870 patients met criteria for colorectal cancer screening; of these 61 percent were screened. Screening was more likely in patients aged 70 to 80 years than in those younger or older. Female gender (relative risk, 0.92; 95 percent confidence interval, 0.9–0.95), Black race (relative risk, 0.92; 95 percent confidence interval, 0.89–0.96), lower income, and infrequent primary care visits were associated with lower likelihood of screening. Of those patients with a positive fecal occult blood test (n = 313), 59 percent received a follow-up barium enema or colonoscopy. Patient-level factors did not predict receipt of a follow-up test. Conclusions The Veterans Health Administration rates for colorectal cancer screening are significantly higher than the national average. However, 41 percent of patients with positive fecal occult blood tests failed to receive follow-up testing. Efforts to measure the quality of colorectal cancer screening programs should focus on the entire diagnostic process. Supported by the Robert Wood Johnson Foundation Clinical Scholars Program (Etzioni), Department of Veterans Affairs (VA) Health Services Research and Development (HSR&D), and National Cancer Institute Colorectal Cancer (CRC) Quality Enhancement Research Initiative (QUERI) Service Directed Research (Project # CRS 02-163). The views expressed in this article are those of the authors and do not necessarily represent the views of the Robert Wood Johnson Foundation or the Department of Veterans Affairs.     

Immunochemical Fecal Occult Blood Test Is Inadequate for Screening Test of Stomach Cancer

The present study was carried out to investigate the diagnostic accuracy of the immunochemical fecal occult blood test in the screening for stomach cancer. In the hospital-based case–control study, the test was positive in 6 (14.3%) subjects with stomach cancer, in 32 (76.2%) subjects with colorectal cancer, and in 10 (7.9%) healthy subjects, respectively, showing a significant difference in detection rate between the subjects with stomach cancer and colorectal cancer (p < 0.01). In the population-based cross-sectional study, detection rate for stomach cancer was 0.13% and 0.15% for negative and positive groups decided by immunochemical fecal occult blood test, indicating no significant difference. These results reveal that the immunochemical fecal occult blood test is inadequate as the screening test for stomach cancer and suggest that examination of the upper digestive tract is unnecessary in cases where the result of fecal occult blood test is positive with no sign of colorectal diseases.     

Screening for colorectal cancer

PURPOSE: Screening for colorectal cancer using a guaiac-based fecal occult blood, or Hemoccult®, test has been demonstrated to reduce colorectal cancer mortality. However, the magnitude of effectiveness is relatively low because of poor sensitivity of the Hemoccult® test. The immunochemical fecal occult blood test has been shown to be much more sensitive than the Hemoccult® test in detecting preclinical colorectal cancer in an asymptomatic population. The purpose of this article is to discuss the validity of the immunochemical fecal occult blood test and the efficacy of a population-based screening program using the test. METHODS: Relevant articles were primarily identified through MEDLINE search. Review was focused on the studies of population screening programs with the immunochemical fecal occult blood test. RESULTS: Sensitivities for colorectal cancer calculated in the same population were reported to be 67 to 89 percent and only 33 to 37 percent for the immunochemical test and Hemoccult® test, respectively. Case-control studies and other observational studies showed that screening programs using the immunochemical fecal occult blood test by hemagglutination reaction would reduce the risk of dying of colorectal cancer by 60 percent or more for those screened annually compared with those unscreened. It was also shown that a screening strategy using the immunochemical fecal occult blood test had the best cost-effectiveness ratio among the methods available. Nearly 5 million persons are currently screened per year in Japan, yielding 0.15 to 0.2 percent colorectal cancer cases among persons with positive fecal occult blood test results. CONCLUSIONS: These results strongly suggest that a screening program with immunochemical fecal occult blood test has promising advantages in terms of effectiveness over programs with the Hemocult® test. More stress is warranted on introduction of immunochemical fecal occult blood testing as a screening test in place of the guaiac fecal occult blood test.     

互补序贯初筛方案对提高大肠癌筛检率的价值

目的：为进一步提高无症状人群普查中大肠癌的检出率,降低普查费用。方法：在既往序贯烘陷血筛检方案（Ｓ方案）、粪隐血与微量白蛋白互补方案（ＳＡ方案）的基础上,探索了互 序贯筛检方案,即烘陷血与微量白蛋白互补→肠道脱落细胞序贯方案（ＳＡＤ方案）。结果：经３９２例已知患者验证,ＳＡＤ方案大肠癌检出敏感性显著高于Ｓ方案（Ｐ〈０．０１）,与ＳＡ方案无显著差异（Ｐ〉０．０１）,特异性显著高于ＳＡ方案（Ｐ〈０．０     

粪便基因甲基化分析对大肠癌的早期诊断价值

目的探讨人粪便中分泌型卷曲相关蛋白(SFRP2)及增生性息肉蛋白(HPP1)基因甲基化分析对患者大肠癌诊断的价值。方法从30例结直肠癌患者、30例结直肠腺瘤患者及30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及HPP1基因甲基化状态。结果大肠癌、腺瘤和正常对照组的SFRP2基因甲基化阳性率分别为66.6%(20/30)、50.0%(15/30)和3.3%(1/30)。大肠癌、腺瘤和正常对照组的HPP1基因甲基化阳性率分别63.3%(19/30)、43.3%(13/30)和6.6%(2/30)。结论 SFRP2和HPP1基因甲基化是大肠癌进展过程中的早期事件。粪便SFRP2和HPP1基因甲基化分析可望成为大肠癌早期无创诊断的新途径。