DSA-FACE技术筛选胰腺癌血清特异糖链的研究
摘要
胰腺癌是一种临床表现隐匿、发展迅速和预后极差的消化系统肿瘤,目前临床上缺乏特异性强的肿瘤标志物。基于DNA测序仪的荧光糖电泳(DSA-FACE)技术是一种能实现高通量、高灵敏度的糖组学检测方法,该技术现正应用于肝脏病学、衰老和乳腺癌等血清糖组学的研究领域。
本研究利用DSA-FACE技术对41例胰腺导管腺癌(PDA)患者血清和100例健康人群血清N-糖链的差异进行比较分析,发现两者之间存在显著性差异。选取在PDA患者组显著高于健康人群组的糖峰中差异最显著的峰14( P <0.001)以及PDA患者组显著低于健康人群组的糖峰中差异最显著的峰6(P <0.001)作为特异性糖链,并以log(p14/p6)为指标进行ROC曲线分析,发现曲线下面积为0.803±0.041,检测的灵敏度为75.6%,特异性为77%。进一步分析发现,峰14和峰6检测胰腺癌早中期的灵敏度可达77.8%,特异性可达80%。以上结果表明,峰14和峰6作为胰腺癌特异性的血清标志物有望成为临床诊断PDA的糖组学指标。
已有研究表明,不同性别间的血清糖谱图存在明显差异,因此我们对不同性别的PDA患者与对照组的糖峰进行比较,结果发现在PDA男性患者中峰15显著高于健康男性,而在女性中没有显著差异,以log(p15/p6)作为指标,ROC曲线下面积为0.865±0.044,对PDA男性患者诊断的灵敏度为92.6%,特异性为80.3%。在PDA女性患者中峰10显著低于健康女性,而在男性中没有显著差异,以峰log(p14/p10)作为指标,ROC曲线下面积为0.925±0.047,诊断PDA女性患者的灵敏度为71.4%,特异性可达到100%。由此可以看出峰15和峰10可能与不同性别当中胰腺癌的发生密切相关,此外这些峰还可分别作为不同性别的胰腺癌临床特异性诊断标志物。此外,我们进一步比较分析了60岁上下的PDA患者和健康人群组的差异,筛选出适合诊断60岁以下和60岁以上PDA患者的糖峰,有望成为个性化医疗的辅助指标,使检测更为精确。
目前临床上使用的胰腺癌肿瘤标志物存在灵敏度和特异性有限、无法实现高通量检测等缺点,本研究利用高通量的DSA-FACE技术筛选出PDA患者特异性的糖峰,其可以作为潜在的胰腺癌诊断标志物。根据不同性别、不同年龄PDA患者的糖链含量存在差异的情况下,筛选出了适合各自诊断的PDA糖组学指标,该结果将会对胰腺癌的预测、诊断及个性化医疗产生影响,并为建立一种可应用于临床的无创、快速、便捷的高通量诊断方法奠定基础。
Pancreatic carcinoma is one of the malignant tumors that has the worst therapeutic effect, because of the often late onset of clinical symptoms, low rate of early diagnosis and poor result of clinical treatment. Till now, the clinicl study is still lack of specific tumor markers. DNA sequencer assisted fluorophore-assisted carbohydrate electrophoresis (DSA-FACE) is a powerful high throughput, high sensitivity detection method for Glycomics and is being used in the research field of serum Glycomics of hepatology, aging and breast cancer.
To find Glycomic specific tumor markers of pancreatic carcinoma, DSA-FACE was performed in 41 patients with pancreatic ductal adenocarcinoma (PDA) and 100 healthy human. The results indicated that there were significant differences in N-linked glycans between PDA patients and healthy persons. Peak 6 and Peak 14 were the most significant peaks which were picked as a new tumor marker of PDA. Taking log (p14/p6) as indicator of the ROC curve analysis, the area under ROC curve was 0.803±0.041 with 75.6% sensitivity and 77% specificity. Further analysis showed that for detection of pancreatic carcinoma early metaphase with peak 14 and peak 6, sensitivity is up to 77.8% and specificity reaches 80%. These results suggested that as the serum markers for pancreatic carcinoma specific clinical diagnosis of PDA, peak 14 and peak 6 might possibly become the Glycomics indicator.
It’s known that there are differences in serum profile between different sexes so we grouped according to the gender to make analysis. It is found that the peak 15 showed significant difference between male patients of PDA and the healthy male but no significant difference in female. Taking log (p15/p6) as indicator, the area under ROC curve is 0.865±0.044 with 92.6% sensitivity and 80.3% specificity. Also, the peak 10 showed significant difference between female patients of PDA and the healthy female but no significant difference in male. Taking log (p14/p10) as indicator, the area under the ROC curve is 0.925±0.047 with 71.4% sensitivity and 100% specificity. So, the peak 15 and peak 10 may be closely related to the occurrence of pancreatic carcinoma of different gender, in addition, these peaks can be also taken as different sex-specific diagnostic markers of pancreatic carcinoma.
Meanwhile, for the age of high incidence of pancreatic carcinoma, the N-glycomic changes in serum were analyzed between PDA and healthy persons of the age at the cutoff 60. the appropriate PDA glycan peaks were checked for the diagnose of PDA between the age over 60 and the age below it. As the specific Glycomics indexes with the cutoff point of 60-year-old for the diagnosis of pancreatic carcinoma, it’s possible that theses results may become an auxiliary index for the personalized health care and make the detection more accurate and useful.
The sensitivity and specificity of the current clinical tumor markers of pancreatic carcinoma is limited and high throughput detection is impossible. In this work, with high-throughput screening technology of DSA-FACE, the specific glycan peaks were founded and it can be used as potential diagnostic markers of pancreatic carcinoma. According to gender, age and many differences cases in PDA, the Glycomics indicator for each diagnosis group were analyzed. The result should be helpful for the forecast of pancreatic carcinoma, diagnosis and personalized medical treatment and might be used as a foundation to establish clinical noninvasive, fast and convenient high-throughput diagnostic methods.
本研究利用DSA-FACE技术对41例胰腺导管腺癌(PDA)患者血清和100例健康人群血清N-糖链的差异进行比较分析,发现两者之间存在显著性差异。选取在PDA患者组显著高于健康人群组的糖峰中差异最显著的峰14( P <0.001)以及PDA患者组显著低于健康人群组的糖峰中差异最显著的峰6(P <0.001)作为特异性糖链,并以log(p14/p6)为指标进行ROC曲线分析,发现曲线下面积为0.803±0.041,检测的灵敏度为75.6%,特异性为77%。进一步分析发现,峰14和峰6检测胰腺癌早中期的灵敏度可达77.8%,特异性可达80%。以上结果表明,峰14和峰6作为胰腺癌特异性的血清标志物有望成为临床诊断PDA的糖组学指标。
已有研究表明,不同性别间的血清糖谱图存在明显差异,因此我们对不同性别的PDA患者与对照组的糖峰进行比较,结果发现在PDA男性患者中峰15显著高于健康男性,而在女性中没有显著差异,以log(p15/p6)作为指标,ROC曲线下面积为0.865±0.044,对PDA男性患者诊断的灵敏度为92.6%,特异性为80.3%。在PDA女性患者中峰10显著低于健康女性,而在男性中没有显著差异,以峰log(p14/p10)作为指标,ROC曲线下面积为0.925±0.047,诊断PDA女性患者的灵敏度为71.4%,特异性可达到100%。由此可以看出峰15和峰10可能与不同性别当中胰腺癌的发生密切相关,此外这些峰还可分别作为不同性别的胰腺癌临床特异性诊断标志物。此外,我们进一步比较分析了60岁上下的PDA患者和健康人群组的差异,筛选出适合诊断60岁以下和60岁以上PDA患者的糖峰,有望成为个性化医疗的辅助指标,使检测更为精确。
目前临床上使用的胰腺癌肿瘤标志物存在灵敏度和特异性有限、无法实现高通量检测等缺点,本研究利用高通量的DSA-FACE技术筛选出PDA患者特异性的糖峰,其可以作为潜在的胰腺癌诊断标志物。根据不同性别、不同年龄PDA患者的糖链含量存在差异的情况下,筛选出了适合各自诊断的PDA糖组学指标,该结果将会对胰腺癌的预测、诊断及个性化医疗产生影响,并为建立一种可应用于临床的无创、快速、便捷的高通量诊断方法奠定基础。
Pancreatic carcinoma is one of the malignant tumors that has the worst therapeutic effect, because of the often late onset of clinical symptoms, low rate of early diagnosis and poor result of clinical treatment. Till now, the clinicl study is still lack of specific tumor markers. DNA sequencer assisted fluorophore-assisted carbohydrate electrophoresis (DSA-FACE) is a powerful high throughput, high sensitivity detection method for Glycomics and is being used in the research field of serum Glycomics of hepatology, aging and breast cancer.
To find Glycomic specific tumor markers of pancreatic carcinoma, DSA-FACE was performed in 41 patients with pancreatic ductal adenocarcinoma (PDA) and 100 healthy human. The results indicated that there were significant differences in N-linked glycans between PDA patients and healthy persons. Peak 6 and Peak 14 were the most significant peaks which were picked as a new tumor marker of PDA. Taking log (p14/p6) as indicator of the ROC curve analysis, the area under ROC curve was 0.803±0.041 with 75.6% sensitivity and 77% specificity. Further analysis showed that for detection of pancreatic carcinoma early metaphase with peak 14 and peak 6, sensitivity is up to 77.8% and specificity reaches 80%. These results suggested that as the serum markers for pancreatic carcinoma specific clinical diagnosis of PDA, peak 14 and peak 6 might possibly become the Glycomics indicator.
It’s known that there are differences in serum profile between different sexes so we grouped according to the gender to make analysis. It is found that the peak 15 showed significant difference between male patients of PDA and the healthy male but no significant difference in female. Taking log (p15/p6) as indicator, the area under ROC curve is 0.865±0.044 with 92.6% sensitivity and 80.3% specificity. Also, the peak 10 showed significant difference between female patients of PDA and the healthy female but no significant difference in male. Taking log (p14/p10) as indicator, the area under the ROC curve is 0.925±0.047 with 71.4% sensitivity and 100% specificity. So, the peak 15 and peak 10 may be closely related to the occurrence of pancreatic carcinoma of different gender, in addition, these peaks can be also taken as different sex-specific diagnostic markers of pancreatic carcinoma.
Meanwhile, for the age of high incidence of pancreatic carcinoma, the N-glycomic changes in serum were analyzed between PDA and healthy persons of the age at the cutoff 60. the appropriate PDA glycan peaks were checked for the diagnose of PDA between the age over 60 and the age below it. As the specific Glycomics indexes with the cutoff point of 60-year-old for the diagnosis of pancreatic carcinoma, it’s possible that theses results may become an auxiliary index for the personalized health care and make the detection more accurate and useful.
The sensitivity and specificity of the current clinical tumor markers of pancreatic carcinoma is limited and high throughput detection is impossible. In this work, with high-throughput screening technology of DSA-FACE, the specific glycan peaks were founded and it can be used as potential diagnostic markers of pancreatic carcinoma. According to gender, age and many differences cases in PDA, the Glycomics indicator for each diagnosis group were analyzed. The result should be helpful for the forecast of pancreatic carcinoma, diagnosis and personalized medical treatment and might be used as a foundation to establish clinical noninvasive, fast and convenient high-throughput diagnostic methods.
引文
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