基于高通量测序技术的膀胱癌转录组分子标记的初步筛选
发布时间：2018-08-16 17:47
【摘要】：目的：基于高通量测序技术、RT-qPCR和免疫组化技术,从转录层面和蛋白表达角度探索膀胱尿路上皮癌发生发展的可能机制,筛选出膀胱肿瘤早期诊断和监测的分子标记。 方法：通过高通量测序技术对一膀胱尿路上皮癌的患者进行转录组测序和生物信息学分析,筛选到候选的差异表达分子,同时收集31对膀胱尿路上皮癌肿瘤组织和癌旁非肿瘤组织样品,采用RT-qPCR验证这些差异表达分子CDH1、VEGFA、PTPRF和CLDN7的表达水平。同时对CDH1和CLDN7进行免疫组化进行检测,探讨这些分子在肿瘤的发生发展中的分子机制和临床意义。 结果：1.CDH1、VEGFA、PTPRF和CLDN7可能成为候选膀胱癌早期诊断和监测的分子标记；CDH1和CLDN7mRNA在肿瘤组织相对癌旁非肿瘤组织表达上调,但差异无统计学意义(P0.05)；2.VEGFA在肿瘤组织中相对癌旁非肿瘤组织表达上调,差异有统计学意义(P0.05);3. CDH1、 VEGFA、PTPRF和CLDN7分别与肿瘤复发关系作多元Logistic回归分析,结果发现PTPRF表达作为肿瘤复发因素进入模型(P=0.002),说明PTPRF和肿瘤复发关系密切；4.CDH1和CLDN7免疫组化显示：CDH1在膀胱肿瘤组织和癌旁非肿瘤组织蛋白表达中未发现有明显统计学意义差异(P0.05),膀胱尿路上皮癌肿瘤组织CLDN7表达上调,具有统计学意义(P0.05)。 结论：1.CLDN7、VEGFA和PTPRF等差异表达基因被纳入可能的膀胱癌分子早期诊断和治疗的分子标记候选集；2.VEGFA可能作为候选的膀胱肿瘤诊断的分子标记,PTPRF可能与膀胱肿瘤的复发关系比较密切。
[Abstract]:Objective: to explore the possible mechanism of bladder urothelial carcinoma from transcriptional level and protein expression based on RT-qPCR and immunohistochemistry, and to screen out the molecular markers for early diagnosis and monitoring of bladder cancer. Methods: transcriptome sequencing and bioinformatics analysis were carried out in a patient with bladder urothelial carcinoma by high-throughput sequencing technique, and candidate differentially expressed molecules were screened. At the same time, 31 samples of bladder urothelial carcinoma were collected, and the expression levels of PTPRF and CLDN7 of CDH1VEGFA-PTPRF and CLDN7 were verified by RT-qPCR. At the same time, CDH1 and CLDN7 were detected by immunohistochemistry to explore the molecular mechanism and clinical significance of these molecules in tumorigenesis and development. Results: 1. The expression of CDH1 and CLDN7 might be the molecular markers for early diagnosis and monitoring of bladder cancer, but the expression of CDH1 and CLDN7mRNA in tumor tissues was significantly higher than that in non-tumor tissues adjacent to tumor tissues, but the difference was not statistically significant (P0.05). The expression of 2.VEGFA was up-regulated in tumor tissues relative to non-tumor tissues adjacent to cancer, and the difference was statistically significant (P0.05). The multivariate Logistic regression analysis showed that the expression of PTPRF as a tumor recurrence factor entered the model (P0. 002), indicating the close relationship between PTPRF and tumor recurrence. 4.CDH1 and CLDN7 immunohistochemical staining showed that there was no significant difference in the expression of CLDN7 between bladder tumor tissue and adjacent non-tumor tissue (P0.05), but the expression of CLDN7 was up-regulated in bladder urothelial carcinoma tissue (P0.05). Conclusion: 1. The differential expression genes such as VEGFA and PTPRF were included in the molecular marker candidate for early diagnosis and treatment of bladder cancer 2.VEGFA may be used as a molecular marker for the diagnosis of bladder cancer, which may be closely related to the recurrence of bladder cancer.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.14
本文编号：2186721
[Abstract]:Objective: to explore the possible mechanism of bladder urothelial carcinoma from transcriptional level and protein expression based on RT-qPCR and immunohistochemistry, and to screen out the molecular markers for early diagnosis and monitoring of bladder cancer. Methods: transcriptome sequencing and bioinformatics analysis were carried out in a patient with bladder urothelial carcinoma by high-throughput sequencing technique, and candidate differentially expressed molecules were screened. At the same time, 31 samples of bladder urothelial carcinoma were collected, and the expression levels of PTPRF and CLDN7 of CDH1VEGFA-PTPRF and CLDN7 were verified by RT-qPCR. At the same time, CDH1 and CLDN7 were detected by immunohistochemistry to explore the molecular mechanism and clinical significance of these molecules in tumorigenesis and development. Results: 1. The expression of CDH1 and CLDN7 might be the molecular markers for early diagnosis and monitoring of bladder cancer, but the expression of CDH1 and CLDN7mRNA in tumor tissues was significantly higher than that in non-tumor tissues adjacent to tumor tissues, but the difference was not statistically significant (P0.05). The expression of 2.VEGFA was up-regulated in tumor tissues relative to non-tumor tissues adjacent to cancer, and the difference was statistically significant (P0.05). The multivariate Logistic regression analysis showed that the expression of PTPRF as a tumor recurrence factor entered the model (P0. 002), indicating the close relationship between PTPRF and tumor recurrence. 4.CDH1 and CLDN7 immunohistochemical staining showed that there was no significant difference in the expression of CLDN7 between bladder tumor tissue and adjacent non-tumor tissue (P0.05), but the expression of CLDN7 was up-regulated in bladder urothelial carcinoma tissue (P0.05). Conclusion: 1. The differential expression genes such as VEGFA and PTPRF were included in the molecular marker candidate for early diagnosis and treatment of bladder cancer 2.VEGFA may be used as a molecular marker for the diagnosis of bladder cancer, which may be closely related to the recurrence of bladder cancer.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.14
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