粪便SDC2基因甲基化检测在结直肠癌辅助诊断中的价值

doi:10.3969/j.issn.1673-8640.2022.04.005

摘要/Abstract

摘要：

目的探讨粪便黏结蛋白聚糖2（SDC2）基因甲基化检测在结直肠癌辅助诊断中的价值。方法选取结直肠癌患者51例（结直肠癌组）、结直肠腺瘤患者22例（腺瘤组）、健康体检者39名（正常对照组）,同时收集所有对象的粪便和血清样本,检测粪便SDC2基因甲基化和血清癌胚抗原（CEA）、糖类抗原19-9（CA19-9）水平;其中有26份粪便样本采用P12全自动样本预处理仪和手工法同时进行样本预处理,比较2种方法预处理后检测结果的相关性。采用Peason相关分析评估2种方法之间的相关性,采用kappa检验分析2种方法的一致性。结果结直肠癌组粪便SDC2基因甲基化阳性率显著高于腺瘤组和正常对照组（P<0.01）。结直肠癌组粪便SDC2基因甲基化阳性率显著高于血清CEA和CA19-9的阳性率（P<0.01）。结直肠癌Ⅰ~Ⅳ期患者之间粪便SDC2基因甲基化阳性率差异均无统计学意义（P>0.05）。结直肠癌Ⅰ~Ⅲ期患者的粪便SDC2基因甲基化阳性率显著高于血清CEA和CA19-9的阳性率（P<0.01）。ROC曲线分析结果显示,粪便SDC2基因甲基化、血清CEA、CA19-9单项检测及联合检测诊断结直肠癌的曲线下面积（AUC）分别为0.965、0.694、0.567、0.976。血清CEA联合粪便SDC2基因甲基化可将结直肠癌阳性率提高至90.2%（46/51）。P12自动化样本预处理仪与手工法预处理样本的检测结果呈正相关（r=0.994,P<0.001）,一致性良好（kappa=0.845,P<0.001）。结论粪便SDC2基因甲基化诊断结直肠癌有较高的敏感性和特异性,在早期结直肠癌的筛查中也有较高的临床价值。

关键词: 黏结蛋白聚糖2, 甲基化, 癌胚抗原, 糖类抗原19-9, 结直肠癌

Abstract:

Objective To investigate the value of methylation determination of fecal adhesin syndecan 2（SDC2） gene in the auxiliary diagnosis of colorectal cancer. Methods Totally,51 patients with colorectal cancer（colorectal cancer group）,22 patients with adenoma（adenoma group） and 39 healthy subjects（healthy control group） were enrolled. The stool and serum samples of all the subjects were collected to determine fecal SDC2 gene methylation and serum carcinoembryonic antigen（CEA） and carbohydrate antigen 19-9（CA19-9）levels. Among them,26 fecal samples were pretreated by P12 automatic sample pretreatment instrument and manual method,and the correlation of the determination results after pretreatment by the 2 methods was compared. Pearson correlation analysis and kappa test were used to evaluate the correlation and consistency between the 2 methods. Results The methylation positive rate of fecal SDC2 gene in colorectal cancer group was higher than those in adenoma group and healthy control group（P<0.01）. In colorectal cancer group,the methylation positive rate of fecal SDC2 gene was higher than those of serum CEA and CA19-9（P<0.01）. There was no statistical significance in methylation positive rate of fecal SDC2 gene between patients with colorectal cancer stage Ⅰ-Ⅳ（P>0.05）. The methylation positive rate of fecal SDC2 gene in patients with colorectal cancer stage Ⅰ-Ⅲ was higher than those of CEA and CA19-9 in serum（P<0.01）. Receiver operating characteristic（ROC） curve analysis showed that the areas under curves（AUC） of the methylation of fecal SDC2 gene,serum CEA,serum CA19-9 and the combined determination for colorectal cancer diagnosis were 0.965,0.694,0.567 and 0.976,respectively. Serum CEA combined with the methylation of fecal SDC2 gene can increase the positive rate of colorectal cancer to 90.2%（46/51）. There was a positive correlation between the determination results of P12 automatic sample pretreatment instrument and manual method（r=0.994,P<0.001）,and the consistency was good（kappa=0.845,P<0.001）. Conclusions The methylation of fecal SDC2 gene has high sensitivity and specificity,which can be used as an auxiliary diagnosis of colorectal cancer,and it also has high clinical value in early colorectal cancer screening.

Key words: Syndecan 2, Methylation, Carcinoembryonic antigen, Carbohydrate antigen 19-9, Colorectal cancer

中图分类号:

R446.1

龚志贇, 江铭磊, 施卫忠, 卢仁泉, 郭林. 粪便SDC2基因甲基化检测在结直肠癌辅助诊断中的价值[J]. 检验医学, 2022, 37(4): 325-329.

GONG Zhiyun, JIANG Minglei, SHI Weizhong, LU Renquan, GUO Lin. Application value of fecal SDC2 gene methylation determination in the auxiliary diagnosis of colorectal cancer[J]. Laboratory Medicine, 2022, 37(4): 325-329.

图/表 5

参考文献 17

[1]	刘宗超, 李哲轩, 张阳, 等. 2020全球癌症统计报告解读[J]. 肿瘤综合治疗电子杂志, 2021, 7(2):1-14.
[2]	夏晨静, 陈志荣. 粪便DNA甲基化检测对结直肠癌诊断及预后评估临床价值研究进展[J]. 临床军医杂志, 2019, 47(6):657-659.
[3]	BRAY F, FERLAY J, SOERJOMATARAM I, et al. Global cancer statistics 2018:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018, 68(6):394-424. DOI URL
[4]	ALEKSANDROVA K, PISCHON T, JENAB M, et al. Combined impact of healthy lifestyle factors on colorectal cancer:a large European cohort study[J]. BMC Med, 2014, 12:168. DOI URL
[5]	LUO H, ZHAO Q, WEI W, et al. Circulating tumor DNA methylation profiles enable early diagnosis,prognosis prediction,and screening for colorectal cancer[J]. Sci Transl Med, 2020, 12(524):eaax7533.
[6]	NIU F, WEN J, FU X, et al. Stool DNA test of methylated Syndecan-2 for the early detection of colorectal neoplasia[J]. Cancer Epidemiol Biomarkers Prev, 2017, 26(9):1411-1419. DOI URL
[7]	中国结直肠癌诊疗规范(2020年版)[J]. 中国实用外科杂志, 2020, 40(6):601-625.
[8]	WEISER M R. AJCC 8th edition:colorectal cancer[J]. Ann Surg Oncol, 2018, 25(6):1454-1455. DOI URL
[9]	柏愚, 杨帆, 马丹, 等. 中国早期结直肠癌筛查及内镜诊治指南(2014年,北京)[J]. 胃肠病学, 2015, 20(6):345-365.
[10]	ZOU H, TAYLOR W R, HARRINGTON J J, et al. High detection rates of colorectal neoplasia by stool DNA testing with a novel digital melt curve assay[J]. Gastroenterology, 2009, 136(2):459-470. DOI URL
[11]	王强, 郑海涛, 丁德祥. 结直肠癌的流行病学和筛查进展[J]. 中国现代医生, 2008, 2(18):103-104.
[12]	BOSCH L J, CARVALHO B, FIJNEMAN R J, et al. Molecular tests for colorectal cancer screening[J]. Clin Colorectal Cancer, 2011, 10(1):8-23. DOI URL
[13]	张丽丽, 吴建新. DNA甲基化——肿瘤产生的一种表观遗传学机制[J]. 遗传, 2006, 28(7):880-885.
[14]	赵慧霞, 李秋文, 董伟伟, 等. 粪便SEPT9基因甲基化检测在结直肠癌早期诊断中的应用研究[J]. 中华临床医师杂志(电子版), 2012, 6(10):2599-2602.
[15]	DENYS A, ALLAIN F. The emerging roles of heparan sulfate 3-O-sulfotransferases in cancer[J]. Front Oncol, 2019, 9:507. DOI URL
[16]	LETOHA T, HUDÁK A, KUSZ E, et al. Contribution of syndecans to cellular internalization and fibrillation of amyloid-β(1-42)[J]. Sci Rep, 2019, 9(1):1393. DOI URL
[17]	ARRINGTON C B, PETERSON A G, YOST H J. Sdc2 and Tbx16 regulate Fgf2-dependent epithelial cell morphogenesis in the ciliated organ of asymmetry[J]. Development, 2013, 140(19):4102-4109. DOI URL

组别	例数	SDC2基因甲基化	CEA	CA19-9
结直肠癌组	51	43（84.3）	25（49.0）^#	17（33.3）^#
Ⅰ期	10	7（70.0）	2（20.0）^#	1（10.0）^#
Ⅱ期	13	10（76.9）	5（38.5）^#	2（15.4）^#
Ⅲ期	11	10（90.9）	6（54.5）^#	5（45.5）^#
Ⅳ期	17	16（94.1）	12（70.6）	9（52.9）
腺瘤组	22	5（22.7）^*	2（9.1）^*	2（9.1）^*
正常对照组	39	3（7.7）^*	1（2.6）^*	1（2.6）^*

组别	例数	SDC2基因甲基化	CEA	CA19-9
结直肠癌组	51	43（84.3）	25（49.0）^#	17（33.3）^#
Ⅰ期	10	7（70.0）	2（20.0）^#	1（10.0）^#
Ⅱ期	13	10（76.9）	5（38.5）^#	2（15.4）^#
Ⅲ期	11	10（90.9）	6（54.5）^#	5（45.5）^#
Ⅳ期	17	16（94.1）	12（70.6）	9（52.9）
腺瘤组	22	5（22.7）^*	2（9.1）^*	2（9.1）^*
正常对照组	39	3（7.7）^*	1（2.6）^*	1（2.6）^*

项目	AUC（95%CI^①）	最佳临界值	敏感性/%	特异性/%	Youden指数
SDC2基因甲基化	0.965（0.912~0.990）	-38.1	88.2	96.7	0.839
CEA	0.694（0.600~0.778）	5.6 ng/mL	45.1	96.7	0.418
CA19-9	0.567（0.470~0.661）	35.0 U/mL	27.5	98.4	0.259
联合检测	0.976（0.928~0.996）	0.40	90.3	98.2	0.885

项目	AUC（95%CI^①）	最佳临界值	敏感性/%	特异性/%	Youden指数
SDC2基因甲基化	0.965（0.912~0.990）	-38.1	88.2	96.7	0.839
CEA	0.694（0.600~0.778）	5.6 ng/mL	45.1	96.7	0.418
CA19-9	0.567（0.470~0.661）	35.0 U/mL	27.5	98.4	0.259
联合检测	0.976（0.928~0.996）	0.40	90.3	98.2	0.885

组别	CEA/（ng/mL）	CEA阳性/例（%）	SDC2基因甲基化阳性/ 例（%）	SDC2基因甲基化+CEA阳性/ 例（%）
结直肠癌组	3.99（2.32~12.5）	25（49.0）	43（84.3）^*	46（90.2）^*
对照组	1.86（1.19~3.03）	2（3.3）	5（8.2）	5（8.2）