粪便DNA中联合SFRP1及SEPT9基因甲基化检测对老年性结直肠癌早期诊断的临床研究
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摘要
目的探讨粪便DNA中联合分泌型卷曲相关蛋白1(SFRP1)及胞裂蛋白9(SEPT9)基因甲基化检测对老年性结直肠癌(CRC)早期诊断的临床价值。方法选取我院从2016年1月至2016年9月确诊的35例年龄>60岁CRC患者为研究组,选取同期35例>60岁正常体检者为对照组。提取CRC患者和正常体检者粪便DNA,采用甲基化特异性PCR(MSP)技术分别检测SFRP1及SEPT9基因的甲基化状态;比较CRC患者单基因和联合双基因甲基化检出率的差异,以及CRC患者联合双基因甲基化检出率与对照组的差异;分析CRC患者SFRP1或SEPT9基因甲基化与临床病理特征(如年龄、性别、肿瘤位置)的关系。结果 CRC患者粪便DNA中SFRP1及SEPT9基因甲基化检出率分别为45.71%(16/35)和51.43%(18/35),均显著高于对照组的8.57%(3/35)和14.29%(5/35),差异均有统计学意义(P<0.01);联合双基因检测结果显示,82.86%(29/35)的CRC患者粪便DNA中至少存在一个基因的甲基化,显著高于单基因检出率(P<0.01),亦高于对照组的17.14%(6/35),差异有统计学意义(P<0.01)。CRC患者SFRP1或SEPT9基因甲基化与年龄、性别、肿瘤位置均无关(P>0.05)。结论粪便DNA中联合SFRP1及SEPT9基因甲基化检测比单基因检测更加敏感,对老年性CRC早期诊断具有重要的应用价值。
Objective To investigate the clinical value of detection of combined secreted frizzled-related proteins 1( SFRP1)and septin 9( SEPT9) gene methylation in stool DNA for early diagnosis of elderly colorectal cancer( CRC). Methods Thirty-five patients with CRC who were aged over 60 years from January 2016 to September 2016 were inrolled in the study. At the same time,35 normal people,who passed the physical examination and were aged over 60 years,were selected as the control group. DNA in the stool of CRC and normal subjects was extracted. Methylation specific PCR( MSP) technique was used to detect the methylation status of SFRP1 and SEPT9 genes respectively. The difference of detection rate of single gene and double gene methylation in CRC patients was compared,and the detection rate of combined double gene methylation in CRC patients was compared with that in normal control group.The correlation between the methylation of SFRP1 or SEPT9 gene in CRC patients and the clinicopathological parameters,such as age,sex,and tumor location,was analyzed. Results The detection rates of methylation of SFRP1 and SEPT9 genes in stool DNA of CRC patients were 45. 71%( 16/35) and 51. 43%( 18/35),respectively,which were significantly higher than those in control group8. 57%( 3/35) and 14. 29%( 5/35),and the differences were statistically significant( P<0. 01). There was at least one gene methylation in the stool DNA of 82. 86%( 29/35) CRC patients,which was significantly higher than the single gene detection rate( P <0. 01),which was also higher than that of the control group 17. 14%( 6/35),the difference was statistically significant( P< 0. 01).The methylation of SFRP1 or SEPT9 genes in CRC patients was not related to age,sex and tumor location( P>0. 05). Conclusion The methylation test of SFRP1 and SEPT9 genes in stool DNA is more sensitive than single gene detection,which is of great value in the early diagnosis of elderly colorectal cancer.
Objective To investigate the clinical value of detection of combined secreted frizzled-related proteins 1( SFRP1)and septin 9( SEPT9) gene methylation in stool DNA for early diagnosis of elderly colorectal cancer( CRC). Methods Thirty-five patients with CRC who were aged over 60 years from January 2016 to September 2016 were inrolled in the study. At the same time,35 normal people,who passed the physical examination and were aged over 60 years,were selected as the control group. DNA in the stool of CRC and normal subjects was extracted. Methylation specific PCR( MSP) technique was used to detect the methylation status of SFRP1 and SEPT9 genes respectively. The difference of detection rate of single gene and double gene methylation in CRC patients was compared,and the detection rate of combined double gene methylation in CRC patients was compared with that in normal control group.The correlation between the methylation of SFRP1 or SEPT9 gene in CRC patients and the clinicopathological parameters,such as age,sex,and tumor location,was analyzed. Results The detection rates of methylation of SFRP1 and SEPT9 genes in stool DNA of CRC patients were 45. 71%( 16/35) and 51. 43%( 18/35),respectively,which were significantly higher than those in control group8. 57%( 3/35) and 14. 29%( 5/35),and the differences were statistically significant( P<0. 01). There was at least one gene methylation in the stool DNA of 82. 86%( 29/35) CRC patients,which was significantly higher than the single gene detection rate( P <0. 01),which was also higher than that of the control group 17. 14%( 6/35),the difference was statistically significant( P< 0. 01).The methylation of SFRP1 or SEPT9 genes in CRC patients was not related to age,sex and tumor location( P>0. 05). Conclusion The methylation test of SFRP1 and SEPT9 genes in stool DNA is more sensitive than single gene detection,which is of great value in the early diagnosis of elderly colorectal cancer.
引文
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[11]Salehi R,Mohammadi M,Emami MH,et al.Methylation pattern of SFRP1 promoter in stool sample is a potential marker for early detection of colorectal cancer[J].Adv Biomed Res,2012,1(1):87.
[12]Wu D,Zhou GP,Ji P,et al.Detection of colorectal cancer using a simplified SEPT9 gene methylation assay is a reliable method for opportunistic screening[J].J Mol Diagn,2016,18(4):535-545.
[13]Xue M,Lai SC,Xu ZP,et al.Noninvasive DNA methylation biomarkers in colorectal cancer:A systematic review[J].Biochim Biophys Acta,2016,1866(1):106-120.
[14]Tanzer M,Balluff B,Distler J,et al.Performance of epigenetic markers SEPT9 and ALX4 in plasma for detection of colorectal precancerous lesions[J/OL].Plo S one,2010[2017-10-27].https://www.ncbi.nlm.nih.gov/pubmed/20140221.
[2]Siegel RL,Miller KD,Fedewa SA,et al.Colorectal cancer statistics,2017[J].CA Cancer J Clin,2017,67(3):177-193.
[3]Kazanets A,Shorstova T,Hilmi K,et al.Epigenetic silencing of tumor suppressor genes:Paradigms,puzzles,and potential[J].Bio Bioph Acta,2016,1865(2):275-288.
[4]Kim J,Kim S.In silico identification of SFRP1 as a hypermethylated gene in colorectal cancers[J].Genomics Inform,2014,12(4):171-180.
[5]Bartak BK,Kalmar A,Peterfia B,et al.Colorectal adenoma and cancer detection based on altered methylation pattern of SFRP1,SFRP2,SDC2,and PRIMA1 in plasma samples[J/OL].Epigenetics,2017[2017-10-25].http://www.tandfonline.com/doi/abs/10.1080/15592294.2017.1356957?journal Code=kepi20.
[6]Song L,Peng X,Li Y,et al.The SEPT9 gene methylation assay is capable of detecting colorectal adenoma in opportunistic screening[J].Epigenomics,2017,9(5):599-610.
[7]Ahmed D,Danielsen SA,Aagesen TH,et al.A tissue-based comparative effectiveness analysis of biomarkers for early detection of colorectal tumors[J/OL].Clin Transl Gastroenterol,2012[2017-10-25].https://www.ncbi.nlm.nih.gov/pubmed/23324654/.
[8]翟爱军,陈洪,王贵齐.三种结直肠癌筛查方法比较研究[J].中国全科医学,2016,19(2):170-173.
[9]Rengucci C,De Maio G,Menghi M,et al.Improved stool DNA integrity method for early colorectal cancer diagnosis.Cancer epidemiology,biomarkers&prevention:a publication of the American Association for Cancer Research,cosponsored by the American Society of Preventive[J].Oncology,2014,23(11):2553-2560.
[10]银广悦,刘创建,周志伟,等.SFRP1及Wi F-1基因启动子甲基化检测在结直肠癌早期筛查中的价值[J].山东医药,2016,56(41):54-56.
[11]Salehi R,Mohammadi M,Emami MH,et al.Methylation pattern of SFRP1 promoter in stool sample is a potential marker for early detection of colorectal cancer[J].Adv Biomed Res,2012,1(1):87.
[12]Wu D,Zhou GP,Ji P,et al.Detection of colorectal cancer using a simplified SEPT9 gene methylation assay is a reliable method for opportunistic screening[J].J Mol Diagn,2016,18(4):535-545.
[13]Xue M,Lai SC,Xu ZP,et al.Noninvasive DNA methylation biomarkers in colorectal cancer:A systematic review[J].Biochim Biophys Acta,2016,1866(1):106-120.
[14]Tanzer M,Balluff B,Distler J,et al.Performance of epigenetic markers SEPT9 and ALX4 in plasma for detection of colorectal precancerous lesions[J/OL].Plo S one,2010[2017-10-27].https://www.ncbi.nlm.nih.gov/pubmed/20140221.