结直肠癌血清IncRNA诊断模型的建立及其对结直肠癌预后监测的临床意义

发布时间：2018-06-05 19:27

本文选题：结直肠癌 + 血清lncNA　；参考：《山东大学》2017年硕士论文

【摘要】：目的:筛选结直肠癌血清中特异表达的lncRNA,建立结直肠癌血清lncRNA诊断模型,并探讨该模型在结直肠癌诊断及预后监测中的临床意义。方法:1.初筛阶段选取6名结直肠癌患者,取其结直肠癌及癌旁正常组织进行基因芯片分析,筛选出结直肠癌与癌旁正常组织间差异表达的lncRNA。结合基因芯片结果及相关文献报道,选取候选lncRNA分子。2.确证阶段采用RT-qPCR技术,首先在80对结直肠癌及癌旁组织中对候选lncRNA分子进行验证,将其中存在显著性差异表达的候选分子在120名结直肠癌患者及120名对照者血清中进行进一步验证,最终确定在组织及血清中均存在显著差异表达的lncRNA分子。将确定的lncRNA分子带入多元logisitic回归分析,建立结直肠癌血清lncRNA诊断模型,采用受试者工作曲线(receiver operating characteristic curve,ROC)分析结直肠癌血清 lncRNA 模型的诊断效能。3.验证阶段另选取120位结直肠癌癌患者和120名对照者血清,采用RT-qPCR方法再次验证上一阶段确定的差异表达分子,并评价血清lncRNA诊断模型在结直肠癌诊断中的诊断价值。同时对这120位结直肠癌患者和120对照者的血清标本CEA进行测定,比较血清lncRNA诊断模型与CEA在结直肠癌诊断中的效能。4.收集并分析验证阶段120名结直肠癌患者的预后信息。采用Kaplan-Meier分析对结直肠癌进行生存曲线分析,通过Cox比例风险模型分析可用于结直肠癌预后监测的独立预后因素。5.分析筛选出的lncRNA分子在组织和血清中表达量的相关性,并检测lncRNA分子的表达稳定性。结果:1.初筛阶段经基因芯片分析,共发现5873个差异表达的lncRNA(≥2个拷贝,p0.05)。根据基因芯片分析结果及相关文献报道,共选取18个lncRNA分子,其中10个候选分子来自芯片结果,8个分子来自文献报道。2.在确证阶段,经RT-qPCR初次验证,4个lncRNA分子(BANCR,NR_026817,NR_029373和NR_034119)在结直肠癌组织及血清中均存在显著差异表达(均p0.001),其中 BANCR 表达上调,NR_026817,NR_029373 和 NR_034119表达均下调。采用ROC曲线评估BANCR,NR_026817,NR_029373和NR_034119诊断结直肠癌的受试者工作曲线下面积(area under the ROC,AUC),分别为0.638,0.708,0.812和0.724。通过Logistic多元回归分析建立基于4个lncRNA分子的结直肠癌血清lncRNA诊断模型,此诊断模型的AUC 为 0.891(95%CI 为 0.844-0.927,灵敏度为 81.67%,特异度为 80.00%)。3.验证阶段用RT-qPCR进行再次验证,结直肠癌血清lncRNA诊断模型AUC为 0.881(95%CI 为 0.833-0.919,灵敏度为 89.17%,特异度 75.83%)。其对TNM I、II、III期结直肠癌的诊断的AUC分别为0.774、0.844和0.949,明显高于 CEA 相应的 0.588、0695 和 0.861,(均 p0.05)。4.Kaplan-Meier分析显示,低表达NR_029373和NR_034119结直肠癌患者较高表达患者的生存率低(p = 0.013和0.044)。多变量Cox回归分析显示,NR_029373和NR_034119可以作为结直肠癌的独立预后指标(p分别为0.013 和 0.038)。5.4 个 lncRNA 分子(BANCR,NR_026817,NR_029373 和 NR_034119)在组织和血清中的表达量呈线性相关,且4个lncRNA分子在室温放置、反复冻融等处理后表达量稳定。结论:1.结直肠癌血清 lncRNA 诊断模型(BANCR,NR_026817,NR_029373 和NR_034119)对结直肠癌早期诊断具有一定的临床意义。2.NR_029373和NR_034119或可作为结直肠癌监测的独立预后指标。
[Abstract]:Objective: to screen the specific expression of lncRNA in the serum of colorectal cancer and to establish a lncRNA diagnostic model for colorectal cancer, and to explore the clinical significance of this model in the diagnosis and prognosis of colorectal cancer. Methods: 1. patients with colorectal cancer were selected at the initial stage of screening, and the colorectal cancer and normal tissues were analyzed by gene chip analysis. The results of the differential expression of lncRNA. gene chip between rectal cancer and adjacent normal tissues and related literature were reported, and RT-qPCR technology was used to select the candidate lncRNA molecule.2. confirmation stage. First of all, the candidate lncRNA molecules were tested in 80 colorectal and para cancer tissues, and there were 120 candidates with significant differences in the expression of the candidate molecules. The sera of the colorectal cancer patients and 120 controls were further confirmed, and the lncRNA molecules with significant differences in the tissues and serum were determined. The lncRNA molecules were taken into multiple logisitic regression analysis, the lncRNA diagnostic model of colorectal cancer serum was established, and the receiver operating characte was used (receiver operating characte). Ristic curve, ROC) analysis of the diagnostic efficiency of the serum lncRNA model of colorectal cancer, 120 cancer patients and 120 control serums were selected from the.3. verification stage, and the RT-qPCR method was used to re verify the differential expression molecules identified in the previous stage, and the diagnostic value of serum lncRNA diagnostic model in the diagnosis of colorectal cancer was evaluated. The serum samples of 120 patients with colorectal cancer and 120 controls were measured to compare the serum lncRNA diagnostic model with the efficacy of CEA in the diagnosis of colorectal cancer and to analyze the prognostic information of 120 colorectal cancer patients at the verification stage. Kaplan-Meier analysis was used to analyze the survival curve of colorectal cancer, and the risk of Cox was proportional to the risk of Cox. The model analysis can be used to determine the correlation of the expression of lncRNA molecules in tissue and serum by independent prognostic factor.5. analysis of colorectal cancer prognosis and to detect the expression stability of lncRNA molecules. Results: 5873 differentially expressed lncRNA (more than 2 copies, P0.05) were detected by gene chip analysis at the 1. initial screening stage. A total of 18 lncRNA molecules were selected from the results of the slice analysis and related literature. 10 of the candidate molecules came from the results of the chip. 8 of the molecules were reported from the literature.2. at the confirmation stage. After the initial verification by RT-qPCR, 4 lncRNA molecules (BANCR, NR_026817, NR_029373 and NR_034119) were significantly different in the tissues and serum of colorectal cancer (p0.0 01) the expression of BANCR, NR_026817, NR_029373 and NR_034119 were all down regulated. The area under the working curve of BANCR, NR_026817, NR_029373 and NR_034119 was used to evaluate the area under the working curve of BANCR, NR_026817, NR_029373 and NR_034119 (area under the). The lncRNA diagnosis model of colorectal cancer serum of RNA molecule was 0.891 (95%CI 0.844-0.927, sensitivity 81.67%, specificity 80%).3. verification stage by RT-qPCR, and the serum lncRNA diagnostic model of colorectal cancer was 0.881 (95%CI for 0.833-0.919, sensitivity 89.17%, specificity 75.83%). The AUC for the diagnosis of TNM I, II, III colorectal cancer was 0.774,0.844 and 0.949 respectively, significantly higher than that of CEA corresponding 0.5880695 and 0.861. (P0.05).4.Kaplan-Meier analysis showed low expression of low expression of NR_029373 and NR_034119 colorectal cancer patients (P = 0.013 and 0.044). Multivariate regression analysis showed that 373 and NR_034119 can be used as an independent prognostic indicator of colorectal cancer (P 0.013 and 0.038).5.4 lncRNA molecules (BANCR, NR_026817, NR_029373 and NR_034119) in a linear correlation in the tissue and serum, and 4 lncRNA molecules are placed at room temperature, and after repeated freezing and thawing, the amount of expression is stable. Conclusion: 1. colorectal cancer blood The clear lncRNA diagnostic model (BANCR, NR_026817, NR_029373 and NR_034119) has certain clinical significance for the early diagnosis of colorectal cancer,.2.NR_029373 and NR_034119, or can be used as an independent prognostic indicator for colorectal cancer monitoring.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.34

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