大肠癌SEPT9基因甲基化检测的性能评估及应用拓展

发布时间：2018-05-13 08:43

本文选题：大肠癌 + 甲基化　；参考：《河北北方学院》2017年硕士论文

【摘要】：SEPT9基因甲基化检测是美国食品药品管理局(Food and Drug Administration,FDA)、欧盟(conformite europeenne,CE)认证和中国食品药品监督管理总局(China Food and Drug Administration,CFDA)批准的用于大肠癌筛查(FDA)和/或早期检测(CE、CFDA)的基因甲基化外周血检测,是世界上第一个获得FDA批准的表观遗传学肿瘤液态活检检测。虽然此检测在大肠癌筛查和早期检测的性能已经得到广泛的验证,但各种影响因素对其性能的影响还未有系统性的研究,其对大肠癌前疾病的检测性能也缺乏足够的数据。本研究旨在探讨患者性别、年龄、癌症分期、癌症部位和算法等因素对此检测性能的影响,并明确此检测对大肠癌前疾病,包括各型腺瘤及息肉的检测性能。本研究以机会性筛查方法共入组428名受试者,其中包括98名大肠癌患者,86名各型腺瘤患者,67名增生性息肉患者和177名无疾病受试者。本研究运用实时荧光定量多聚酶链式反应(qRT-PCR)对SEPT9基因启动子区域的甲基化水平进行检测,试验采用ABI7500 Fast DX荧光定量PCR仪和Epi proColon 2.0 CE试剂盒进行检测。所有血液样品采集、运输、处理、血浆提取、DNA保存、亚硫酸盐转化、PCR、数据分析和判读等操作均遵循Epi proColon 2.0 CE试剂盒说明书进行。统计学分析和制图采用GraphPad PRISM 5.0软件完成。研究结果表明:1/3算法在大肠癌、腺瘤、增生性息肉的阳性检出率(positive detection rate,PDR)均高于2/3算法,然而,其在正常人中的假阳性率也高于2/3算法。不论采取何种算法,SEPT9检测对0期和I期大肠癌的阳性检出率显著低于对II~IV期大肠癌的阳性检出率。然而,SEPT9检测对0期和I期大肠癌的检出率均超过50%,说明其可以应用于大肠早癌筛查。60岁以上正常人群的阳性率显著高于小于60岁的人群,而在小于60岁人群中,SEPT9检测的阳性率随年龄组增高而逐渐递增。与此相对,大肠癌人群的阳性检出率在各年龄组间无显著性差异。进一步研究显示,大肠癌患者的阳性检出率在各大肠癌分期均未显示性别方面的差异,在结直肠各个部位,包括回盲部、升结肠、横结肠、降结肠、乙状结肠、直肠,SEPT9检测的阳性检出率也没有发现显著性差异。针对腺瘤的分型检测显示,若使用1/3算法,锯齿状腺瘤(serrated adenoma,SA)的PDR为27.8%,管状腺瘤(tubular adenoma,TA)的PDR为28.7%,管状绒毛腺瘤(tubulovillous adenoma,TVA)的PDR为53.7%,绒毛状腺瘤(villous adenoma,VA)的PDR为83.3%,表明PDR与腺瘤中绒毛成分的含量呈正相关。对TVA和VA的高阳性检出率说明SEPT9检测可用于癌前进展期腺瘤(advanced adenoma,AA)的筛查。我们的研究说明,算法、癌症分期和受检者年龄对SEPT9检测的性能有影响,而性别和癌症部位对检测性能没有影响。SEPT9检测对含绒毛成分的腺瘤有较高的灵敏度,而对增生性息肉的检出率较低,因此,它可作为进展期腺瘤及早期大肠癌的血液筛查工具。
[Abstract]:The SEPT9 gene methylation test was approved by the Food and Drug Administration (FDA), the Food and Drug Administration (and Drug), the European Union (EU), and the China Food and Drug Administration (CFDA-approved) for the screening of colorectal cancer and / or early detection for peripheral blood methylation. It is the first liquid biopsy of epigenetic tumors approved by FDA in the world. Although the performance of this test has been widely verified in colorectal cancer screening and early detection, the influence of various factors on its performance has not been systematically studied, and the detection performance of colorectal cancer precancerous disease is also lack of sufficient data. The purpose of this study was to investigate the effect of sex, age, stage, location and algorithm of cancer on the detection of precancerous diseases, including adenomas and polyps. In this study, a total of 428 subjects were enrolled by opportunistic screening, including 98 patients with colorectal cancer and 86 patients with adenoma of various types, 67 patients with proliferative polyps and 177 subjects without disease. In this study, the methylation level of promoter region of SEPT9 gene was detected by real-time fluorescence quantitative polymerase chain reaction (QRT-PCR), and ABI7500 Fast DX fluorescence quantitative PCR assay and Epi proColon 2.0 CE kit were used to detect the methylation level. All blood samples were collected, transported, processed, plasma extracted and preserved, sulfite transformed into PCRs, data analysis and interpretation were performed according to the instructions of Epi proColon 2.0 CE kit. Statistical analysis and cartography were completed by GraphPad PRISM 5.0 software. The results show that the positive detection positive rate of positive polyps in colorectal cancer, adenoma and proliferative polyps is higher than that in the control group. However, the false positive rate is also higher in the normal population than that in the control group. The positive rate of SEPT9 in stage 0 and stage I colorectal cancer was significantly lower than that in II~IV stage of colorectal cancer. However, the detection rate of stage 0 and stage I colorectal cancer by SEPT9 was more than 50, which indicated that the positive rate of SEPT9 was significantly higher than that of those aged less than 60 years in the normal population over 60 years of age. The positive rate of SEPT9 increased with the increase of age group. In contrast, there was no significant difference in the positive rate of colorectal cancer among different age groups. Further studies showed that there was no gender difference in the positive rate of colorectal cancer in all stages of colorectal cancer, including ileocecal, ascending, transverse, descending, sigmoid colon, There was no significant difference in the positive rate of SEPT9 in rectal cancer. For adenoma typing, if you use the 1 / 3 algorithm, The PDR of serrated adenoma was 27.8, the PDR of tubular adenoma was 28.7, the PDR of tubulovillous adenoma was 53.7 and the PDR of villous adenoma was 83.3, which indicated that PDR was positively correlated with the content of chorionic component in tubulovillous adenoma. The high positive rates of TVA and VA suggest that SEPT9 can be used to screen advanced adenoma. Our study shows that the algorithm, cancer stage and age of the person tested have influence on the performance of SEPT9 detection, but gender and cancer location have no effect on the detection performance. SEPT9 detection has high sensitivity to adenoma containing villi. But the detection rate of proliferative polyps is low, so it can be used as a blood screening tool for advanced adenoma and early colorectal cancer.
【学位授予单位】：河北北方学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.34

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