血小板反应蛋白-4在前列腺癌中的表达方式及生物学意义的研究
发布时间：2018-07-29 17:11
【摘要】：目的 探讨THBS4异常表达在前列腺癌浸润和转移中的作用,并进一步明确其在异种移植动物模型中的表达机制。 方法 通过cDNA基因芯片检测前列腺癌根治术后癌组织和癌旁组织中差异表达基因谱。我们通过文献检索、网络软件和生物信息学在线工具,对基因芯片筛选出的差异表达基因进行基因本体注释,了解差异表达基因的生物学功能、参与构成的细胞成份、参与调控的信号转导通路以及蛋白-蛋白间相互作用的关系网,从而明确前列腺癌组织和癌旁组织在生物学进程上的异同,并选定THBS4作为研究对象。为了验证基因芯片的结果,通过RT-PCR和免疫组化检测方法,利用10例新鲜前列腺癌和癌旁组织进行检测。为了深入探讨THBS4基因在前列腺癌中的表达与作用机制,通过免疫组化技术检测了60例前列腺癌石蜡组织标本,结合临床资料,进行Kaplan-meier生存分析和COX回归风险因素分析。最后,通过显微外科技术建立前列腺癌异种移植模型,通过常规HE染色、冰冻切片、免疫组化检测等对模型进行鉴定,然后进一步检测THBS4在前列腺癌和增生异种移植模型中的表达情况,对前期实验结果进行验证。 结果 前列腺癌与癌旁组织在基因表达上存在显著差异。THBS4基因在癌组织中较癌旁组织表现出一致的高表达。新鲜前列腺癌组织RT-PCR以及免疫组化证实了THBS4在癌组织中高表达。THBS4在前列腺癌间质组织和上皮细胞中表达,并且其表达与Gleason评分、病理类型及分期相关。结合COX回归分析显示：前列腺癌组织中THBS4高表达与转移导致的患者生存期下降显著相关。我们采用显微外科技术建立患者来源的前列腺癌和前列腺增生NOD/SCID小鼠肾包膜下异种移植模型。THBS4在良性前列腺增生小鼠异种模型中未见明显表达,在前列腺癌模型中的表达与原前列腺癌组织表达相近。典型的高表达出现在上皮细胞,靠近肾脏的肿瘤边缘间质中THBS4高表达。 结论 我们通过前列腺癌根治术后癌组织和癌旁组织基因芯片获得了转录水平的差异表达基因普,表明前列腺癌间质在前列腺癌的发生发展中起到了重要作用。我们成功建立了NOD/SCID小鼠肾包膜下患者来源前列腺癌异种移植模型和NOD/SCID小鼠肾包膜下患者来源前列腺增生异种移植模型,并获得了较高的成瘤率。我们认为THBS4可以作为不同恶性程度或者不同阶段的前列腺癌细胞发生上皮-间质转化的标记物,结合Gleason评分、病理类型等可以作为前列腺癌的预后判定指标。
[Abstract]:Objective to investigate the role of abnormal expression of THBS4 in the invasion and metastasis of prostate cancer and to further clarify its expression mechanism in xenotransplantation animal models. Methods cDNA gene microarray was used to detect differentially expressed gene profiles in cancer tissues and paracancerous tissues after radical prostatectomy. Through literature retrieval, network software and online bioinformatics tools, we annotate the differentially expressed genes screened by gene chips, understand the biological functions of differentially expressed genes, and participate in the cellular components. The signal transduction pathway involved in the regulation and the protein-protein interaction network were involved in the study, so as to identify the differences and similarities between prostate cancer tissues and para-cancerous tissues in biological processes, and THBS4 was selected as the research object. In order to verify the results of the microarray, 10 cases of fresh prostate cancer and adjacent tissues were detected by RT-PCR and immunohistochemistry. In order to investigate the expression and mechanism of THBS4 gene in prostate cancer, 60 paraffin tissue specimens of prostate cancer were detected by immunohistochemical technique. Kaplan-meier survival analysis and COX regression risk factor analysis were performed in combination with clinical data. Finally, the xenotransplantation model of prostate cancer was established by microsurgical technique. The model was identified by routine HE staining, frozen sections and immunohistochemistry. Then, the expression of THBS4 in prostate cancer and proliferative xenotransplantation model was detected, and the experimental results were verified. Results there was a significant difference in gene expression between prostate cancer and paracancerous tissues. THBS4 gene expression in cancer tissues was consistent with that in paracancerous tissues. The high expression of THBS4 in stromal tissue and epithelial cells of prostate cancer was confirmed by RT-PCR and immunohistochemistry, and the expression was correlated with Gleason score, pathological type and stage. Combined with COX regression analysis, the high expression of THBS4 in prostate cancer tissues was significantly correlated with the decrease of survival time caused by metastasis. The subcapsular xenotransplantation model of prostate cancer and prostatic hyperplasia in NOD/SCID mice was established by microsurgical technique. The expression of THBS4 was not found in the xenogeneic model of benign prostatic hyperplasia (BPH) mice. The expression in prostate cancer model was similar to that in proto-prostate cancer tissue. Typically, high expression of THBS4 occurs in epithelial cells, adjacent to the renal marginal stroma. Conclusion the differentially expressed genes were obtained by microarray of cancer tissues and paracancerous tissues after radical prostatectomy, indicating that stroma plays an important role in the development of prostate cancer. We successfully established the xenotransplantation model of prostate cancer derived from subcapsular prostate cancer in NOD/SCID mice and the xenotransplantation model of prostatic hyperplasia derived from subcapsular patients in NOD/SCID mice, and obtained a high tumorigenesis rate. We believe that THBS4 can be used as a marker of epithelial-interstitial transformation in prostate cancer cells with different malignant degrees or stages, combined with Gleason score, pathological type and so on, which can be used as a prognostic index of prostate cancer.
【学位授予单位】：天津医科大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.25
本文编号：2153402
[Abstract]:Objective to investigate the role of abnormal expression of THBS4 in the invasion and metastasis of prostate cancer and to further clarify its expression mechanism in xenotransplantation animal models. Methods cDNA gene microarray was used to detect differentially expressed gene profiles in cancer tissues and paracancerous tissues after radical prostatectomy. Through literature retrieval, network software and online bioinformatics tools, we annotate the differentially expressed genes screened by gene chips, understand the biological functions of differentially expressed genes, and participate in the cellular components. The signal transduction pathway involved in the regulation and the protein-protein interaction network were involved in the study, so as to identify the differences and similarities between prostate cancer tissues and para-cancerous tissues in biological processes, and THBS4 was selected as the research object. In order to verify the results of the microarray, 10 cases of fresh prostate cancer and adjacent tissues were detected by RT-PCR and immunohistochemistry. In order to investigate the expression and mechanism of THBS4 gene in prostate cancer, 60 paraffin tissue specimens of prostate cancer were detected by immunohistochemical technique. Kaplan-meier survival analysis and COX regression risk factor analysis were performed in combination with clinical data. Finally, the xenotransplantation model of prostate cancer was established by microsurgical technique. The model was identified by routine HE staining, frozen sections and immunohistochemistry. Then, the expression of THBS4 in prostate cancer and proliferative xenotransplantation model was detected, and the experimental results were verified. Results there was a significant difference in gene expression between prostate cancer and paracancerous tissues. THBS4 gene expression in cancer tissues was consistent with that in paracancerous tissues. The high expression of THBS4 in stromal tissue and epithelial cells of prostate cancer was confirmed by RT-PCR and immunohistochemistry, and the expression was correlated with Gleason score, pathological type and stage. Combined with COX regression analysis, the high expression of THBS4 in prostate cancer tissues was significantly correlated with the decrease of survival time caused by metastasis. The subcapsular xenotransplantation model of prostate cancer and prostatic hyperplasia in NOD/SCID mice was established by microsurgical technique. The expression of THBS4 was not found in the xenogeneic model of benign prostatic hyperplasia (BPH) mice. The expression in prostate cancer model was similar to that in proto-prostate cancer tissue. Typically, high expression of THBS4 occurs in epithelial cells, adjacent to the renal marginal stroma. Conclusion the differentially expressed genes were obtained by microarray of cancer tissues and paracancerous tissues after radical prostatectomy, indicating that stroma plays an important role in the development of prostate cancer. We successfully established the xenotransplantation model of prostate cancer derived from subcapsular prostate cancer in NOD/SCID mice and the xenotransplantation model of prostatic hyperplasia derived from subcapsular patients in NOD/SCID mice, and obtained a high tumorigenesis rate. We believe that THBS4 can be used as a marker of epithelial-interstitial transformation in prostate cancer cells with different malignant degrees or stages, combined with Gleason score, pathological type and so on, which can be used as a prognostic index of prostate cancer.
【学位授予单位】：天津医科大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.25
【参考文献】
相关期刊论文 前2条
1 叶定伟;李长岭;;前列腺癌发病趋势的回顾和展望[J];中国癌症杂志;2007年03期
2 Sathiya Maran;Yeong Yeh Lee;Shuhua Xu;Nur-Shafawati Rajab;Norhazrini Hasan;Syed Hassan Syed Abdul Aziz;Noorizan Abdul Majid;Bin Alwi Zilfalil;;Gastric precancerous lesions are associated with gene variants in Helicobacter pylori -susceptible ethnic Malays[J];World Journal of Gastroenterology;2013年23期
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