【作者】 董德鑫；

【导师】 李汉忠； 纪志刚； 毛全宗；

【作者基本信息】 中国协和医科大学 ， 泌尿外科， 2010， 博士

【摘要】 背景：肾透明细胞癌早期常常无明显临床症状,临床缺乏准确、敏感的肾透明细胞癌诊断和监测用肿瘤标记物。肿瘤标志物大多为异常表达的蛋白质,通过对蛋白质的动态分析,可以探知疾病早期最微小的指标和征兆。寻找肿瘤中差异表达蛋白质是蛋白质组学研究的重点,因此蛋白质组学为肾癌标志物的筛选提供了理想的技术平台。第一部分目的：应用WCX磁珠联合MALDI-TOF MS技术对肾透明细胞癌进行血清差异蛋白研究,寻找显著性表达的差异蛋白。方法：应用WCX磁珠联合MALDI-TOF MS技术对肾透明细胞癌血清进行差异蛋白研究,筛选出肾透明细胞癌不同分组之间多个血清差异蛋白,并应用遗传算法建立诊断模型。结果：1.首次应用WCX磁珠联合MALDI-TOF-MS技术对肾透明细胞癌进行血清差异蛋白研究,该技术具有很好的重复性、可行性和可信性。2.首次应用WCX磁珠联合MALDI-TOF-MS技术筛选出肾透明细胞癌不同分组之间多个具有统计学意义的血清差异蛋白(P＜0.05)。(1)肾透明细胞癌术前组与非肿瘤对照组之间有显著性差异的蛋白峰5个,最显著差异的2个蛋白峰为M=2682.68,2864.38(P=0.000166,0.03)(2)肾透明细胞癌术前组与肾血管平滑肌脂肪瘤组之间有显著性差异的蛋白峰6个,最显著差异的2个蛋白峰为M=2683.14,1331.32(P=0.00203,0.00388)(3)肾透明细胞癌局限组与浸润转移组之间有显著性差异的蛋白峰2个,M=9293.53,4646.76(P=0.0212,0.0389)。(4)肾透明细胞癌术前组与术后组之间有显著性差异的蛋白峰10个,最显著差异的2个蛋白峰为M=3885.32,7769.69(P=0.00231,0.0167)。3.首次应用遗传算法建立肾透明细胞癌不同分组的诊断模型,诊断识别能力分别为94.45%,100%,91.67%和100%。结论：1.首次应用WCX磁珠联合MALDI-TOF MS技术进行肾透明细胞癌血清和尿液差异蛋白研究,该技术具有很好的重复性、可行性和可信性。2.首次筛选出肾透明细胞癌与比较组之间多个具有统计学意义的血清差异蛋白(P＜0.05)。3.首次应用遗传算法建立肾透明细胞癌血清和尿液差异蛋白的诊断模型,识别能力高。第二部分目的：应用WCX磁珠联合MALDI-TOF MS技术对肾透明细胞癌进行尿液差异蛋白研究,寻找显著性表达的差异蛋白。方法：应用WCX磁珠联合MALDI-TOF MS技术对肾透明细胞癌进行尿液差异蛋白研究,筛选出肾透明细胞癌不同分组之间多个尿液差异蛋白,并应用遗传算法建立诊断模型。结果：1.首次应用WCX磁珠联合MALDI-TOF MS技术对肾透明细胞癌进行尿液差异蛋白研究,该技术具有很好的重复性、可行性和可信性。2.首次应用WCX磁珠联合MALDI-TOF MS技术筛选出肾透明细胞癌不同分组之间多个具有统计学意义的尿液差异蛋白(P＜0.05)。(1)肾透明细胞癌术前组与正常对照组之间有显著性差异的蛋白峰1个,差异蛋白峰为M=2221.71(P=0.0304)。(2)肾透明细胞癌组与肾血管平滑肌脂肪瘤组之间有显著性差异的蛋白峰41个,最显著差异的2个蛋白峰为M=4270.15,2251.62(P=0.0167,P=0.0167)。3.首次应用遗传算法建立肾透明细胞癌不同分组的诊断模型,识别能力分别为100%和100%。结论：1.首次应用WCX磁珠联合MALDI-TOF MS技术进行肾透明细胞癌血清和尿液差异蛋白研究,该技术具有很好的重复性、可行性和可信性。2.首次筛选出肾透明细胞癌与比较组之间多个具有统计学意义的血清差异蛋白(P＜0.05)。3.首次应用遗传算法建立肾透明细胞癌血清和尿液差异蛋白的诊断模型,识别能力高。更多还原

【Abstract】 BackgroundThe early symptoms of renal clear cell carcinoma are often not obvious, and we are short of accurate and sensitive tumor marker for diagnosis and monitoring of renal clear cell carcinoma in clinical practice. Most tumor markers are proteins with abnormal expression. By means of dynamic analysis of proteins, we can detect the minor index and early symptoms of disease. The emphasis of proteomics is to search for proteins differentially expressed in cancer. Therefore proteomics provide an ideal technology platform for the screening tumor marker of renal cell carcinoma.Part IObjectiveThe application of WCX magnetic beads combined MALDI-TOF MS in detecting differentially expressed proteins in serum of renal clear cell carcinoma.MethodsWCX magnetic beads combined MALDI-TOF MS are applied in order to screen differentially expressed proteins in serum of renal clear cell carcinoma. Genetic algorithm (GA) is applied to establish diagnosis model.Results1. We are the first to apply WCX magnetic beads combined MALDI-TOF MS in screening differentially expressed proteins in serum of renal clear cell carcinoma. And the technique has good reproducibility, feasibility and credibility.2. We are the first to find many proteins statistically differentially expressed in serum with application of WCX magnetic beads combined MALDI-TOF-MS between the group of renal clear cell carcinoma and other groups (P＜0.05).(1) There are five protein peaks statistically differentially expressed between the group of preoperative renal clear cell carcinoma and the control group of non-tumor. The two protein peaks with the most significant difference are M=2682.68 and M=2864.38 (P=0.000166, P=0.03) (2) There are six protein peaks statistically differentially expressed between the group of preoperative renal clear cell carcinoma and the group of renal angiomyolipoma. The two protein peaks with the most significant difference are M=2683.14 and M=1331.32 (P=0.00203, P=0.00388).(3) There are two protein peaks statistically differentially expressed between the limited group and invasion and metastasis group of preoperative renal clear cell carcinoma. The two protein peaks with the most significant difference are M=9293.53 and M=4646.76 (P=0.0212, P=0.0389).(4) There are ten protein peaks statistically differentially expressed between the preoperative group and postoperative group of renal clear cell carcinoma. The two protein peaks with the most significant difference are M=3885.32 and M=7769.69 (P=0.00231,P=0.0167).3. We are the first to establish diagnosis models of renal clear cell carcinoma by means of genetic algorithm, and the recognition capabilities are 94.45%,100%,91.67% and 100%, respectively.Conclusions1. We are the first to apply WCX magnetic beads combined MALDI-TOF MS in screening differentially expressed proteins in serum of renal clear cell carcinoma. And the technique has good reproducibility, feasibility and credibility.2. We are the first to find many proteins statistically differentially expressed in serum with application of WCX magnetic beads combined MALDI-TOF-MS between the group of renal clear cell carcinoma and other groups (P＜0.05).3. We are the first to establish diagnosis models of renal clear cell carcinoma by means of genetic algorithm, and the recognition capabilities are high.Part IIObjectiveThe application of WCX magnetic beads combined MALDI-TOF MS in detecting differentially expressed proteins in urine of renal clear cell carcinoma. MethodsWCX magnetic beads combined MALDI-TOF MS are applied in order to screen differentially expressed proteins in urine of renal clear cell carcinoma. Genetic algorithm (GA) is applied to establish diagnosis model.Results1. We are the first to apply WCX magnetic beads combined MALDI-TOF MS in screening differentially expressed proteins in urine of renal clear cell carcinoma. And the technique has good reproducibility, feasibility and credibility.2. We are the first to find many proteins statistically differentially expressed in urine with application of WCX magnetic beads combined MALDI-TOF-MS between the group of renal clear cell carcinoma and other groups (P<0.05).(1) There are one protein peak statistically differentially expressed between the group of preoperative renal clear cell carcinoma and the normal control group. The protein peaks with significant difference is M=2221.71(P=0.0304).(2) There are forty-one protein peaks statistically differentially expressed between the group of renal clear cell carcinoma and the group of renal angiomyolipoma. The two protein peaks with the most significant difference are M=4270.15 and M=2251.62 (P=0.0167,P=0.0167).3. We are the first to establish diagnosis models of renal clear cell carcinoma by means of genetic algorithm, and the recognition capabilities are 100%and 100%.Conclusions1. We are the first to apply WCX magnetic beads combined MALDI-TOF MS in screening differentially expressed proteins in urine of renal clear cell carcinoma. And the technique has good reproducibility, feasibility and credibility.2. We are the first to find many proteins statistically differentially expressed in urine with application of WCX magnetic beads combined MALDI-TOF-MS between the group of renal clear cell carcinoma and other groups (P<0.05).3. We are the first to establish diagnosis models of renal clear cell carcinoma by means of genetic algorithm, and the recognition capabilities are high.更多还原