【摘要】：目的 探討THBS4異常表達在前列腺癌浸潤和轉(zhuǎn)移中的作用,并進一步明確其在異種移植動物模型中的表達機制。 方法 通過cDNA基因芯片檢測前列腺癌根治術(shù)后癌組織和癌旁組織中差異表達基因譜。我們通過文獻檢索、網(wǎng)絡軟件和生物信息學在線工具,對基因芯片篩選出的差異表達基因進行基因本體注釋,了解差異表達基因的生物學功能、參與構(gòu)成的細胞成份、參與調(diào)控的信號轉(zhuǎn)導通路以及蛋白-蛋白間相互作用的關(guān)系網(wǎng),從而明確前列腺癌組織和癌旁組織在生物學進程上的異同,并選定THBS4作為研究對象。為了驗證基因芯片的結(jié)果,通過RT-PCR和免疫組化檢測方法,利用10例新鮮前列腺癌和癌旁組織進行檢測。為了深入探討THBS4基因在前列腺癌中的表達與作用機制,通過免疫組化技術(shù)檢測了60例前列腺癌石蠟組織標本,結(jié)合臨床資料,進行Kaplan-meier生存分析和COX回歸風險因素分析。最后,通過顯微外科技術(shù)建立前列腺癌異種移植模型,通過常規(guī)HE染色、冰凍切片、免疫組化檢測等對模型進行鑒定,然后進一步檢測THBS4在前列腺癌和增生異種移植模型中的表達情況,對前期實驗結(jié)果進行驗證。 結(jié)果 前列腺癌與癌旁組織在基因表達上存在顯著差異。THBS4基因在癌組織中較癌旁組織表現(xiàn)出一致的高表達。新鮮前列腺癌組織RT-PCR以及免疫組化證實了THBS4在癌組織中高表達。THBS4在前列腺癌間質(zhì)組織和上皮細胞中表達,并且其表達與Gleason評分、病理類型及分期相關(guān)。結(jié)合COX回歸分析顯示：前列腺癌組織中THBS4高表達與轉(zhuǎn)移導致的患者生存期下降顯著相關(guān)。我們采用顯微外科技術(shù)建立患者來源的前列腺癌和前列腺增生NOD/SCID小鼠腎包膜下異種移植模型。THBS4在良性前列腺增生小鼠異種模型中未見明顯表達,在前列腺癌模型中的表達與原前列腺癌組織表達相近。典型的高表達出現(xiàn)在上皮細胞,靠近腎臟的腫瘤邊緣間質(zhì)中THBS4高表達。 結(jié)論 我們通過前列腺癌根治術(shù)后癌組織和癌旁組織基因芯片獲得了轉(zhuǎn)錄水平的差異表達基因普,表明前列腺癌間質(zhì)在前列腺癌的發(fā)生發(fā)展中起到了重要作用。我們成功建立了NOD/SCID小鼠腎包膜下患者來源前列腺癌異種移植模型和NOD/SCID小鼠腎包膜下患者來源前列腺增生異種移植模型,并獲得了較高的成瘤率。我們認為THBS4可以作為不同惡性程度或者不同階段的前列腺癌細胞發(fā)生上皮-間質(zhì)轉(zhuǎn)化的標記物,結(jié)合Gleason評分、病理類型等可以作為前列腺癌的預后判定指標。
[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.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R737.25

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