FoxM1在前列腺癌上皮间质转化过程中作用的研究

发布时间：2018-04-03 08:18

本文选题：前列腺癌　切入点：FoxM1　出处：《中国人民解放军医学院》2015年博士论文

【摘要】：前列腺癌是威胁中老年男性生命的恶性肿瘤。近年在我国呈递增趋势。前列腺癌治疗方案通常取决于是否发生周围组织浸润和转移。因此对前列腺癌的早期诊断和转移机制的研究具有重要意义。FoxM1参与肿瘤致病的多个环节；上皮-间质转化是肿瘤转移过程中的重要环节。剪切波弹性成像是鉴别肿瘤组织良恶性的重要方法。基于上述研究背景,本研究主要通过检测FoxM1在前列腺癌组织中的表达,探讨FoxM1参与前列腺癌EMT转移机制及影响因素,并通过剪切波弹性成像初步探讨人前列腺癌异种移植瘤弹性,并分析病理组织学基础。方法：1FoxM1在前列腺癌组织及细胞表达的鉴定。检测FoxM1蛋白及基因亚型在前列腺癌、良性前列腺增生组织及前列腺癌细胞系中的表达,分析FoxM1表达与前列腺癌病理Gleason评分的关系。检测FoxM1 3个亚型FoxM1a、FoxM1b、 FoxM1c在前列腺癌组织和细胞系中的表达。2FoxM1参与前列腺癌细胞系EMT的体外实验。建立前列腺癌细胞EMT模型,检测FoxM1与EMT相关分子E-cadherin、vimentin、Slug表达变化,评估细胞运动迁移能力;二甲双胍和低氧环境处理前列腺癌细胞,检测FoxM1表达及EMT情况,评估细胞增殖、运动和迁移能力。3FoxM1参与前列腺癌转移的动物实验。构建携载GFP的FoxM1 shRNA慢病毒载体,建立稳定表达GFP和FoxM1 shRNA前列腺癌细胞株；建立裸鼠移植瘤模型,检测移植瘤生长增殖及转移情况。4.剪切波弹性成像评估移植瘤弹性.建立前列腺癌异种移植瘤模型,灰阶超声测量不同生长时间(6w、8w、10w)瘤体大小,剪切波弹性成像检测瘤体弹性。分析瘤体组织内纤维成分,以及瘤体内胶原分型,检测瘤体组织内α-SMA表达。建立FoxM1敲低移植瘤模型,生长8w时检测上述指标.结果：1.FoxM1在前列腺癌组织和良性前列腺增生组织表达存在差异,在前列腺癌细胞系中均表达,在Gleason评分≥7的癌组织中表达明显高于Gleason7的癌组织(p0.05)。FoxM1c亚型在前列腺癌组织和细胞中表达水平较高。2.TGF-p诱导前列腺癌细胞系EMT,E-cadherin下调,vimentin和Slug上调,细胞运动迁移能力增强,FoxM1敲低后,EMT过程逆转。二甲双胍处理细胞,内源性FoxM1表达下降,细胞增殖及运动迁移能力减弱；低氧环境下细胞内FoxM1表达增加,促进前列腺癌细胞EMT,细胞运动迁移能力增加。3.成功构建携载GFP及FoxM1 shRNA的慢病毒载体,建立FoxM1敲低的前列腺癌移植模型。FoxM1敲低后成瘤能力弱,瘤体生长慢,转移灶较对照组明显减少。4.接种6w.8w.10w瘤体体积分别为832.58±14298mm3,1862.76±271.8mm3和2764.2±255.93mm3(p0.05)；瘤体杨氏模量分别为22.11±2.45KPa,44.89±3.02KPa和57.41±3.98KPa(p0.05)。在较硬瘤体中胶原纤维较多,胶原以Ⅰ型为主,且a-SMA表达较多。FoxM1敲低后瘤体较对照组生长缓慢,接种8w时瘤体大小分别为813.7686±111.1932mm3和1921.297±258.1513mm3(p0.05);杨氏模量分别为25.46±3.48KPa和46.94±5.32KPa(p0.05),在Fox M1敲低组胶原纤维成分较少,Ⅰ型胶原较少,且α^ A表达较少。结论FoxM1在前列腺癌组织中表达量增加,且与前列腺癌Gleason评分密切相关。FoxM1在前列腺癌EMT过程中具有重要作用,有望成为前列腺癌治疗新的靶点。剪切波弹性成像能够评价前列腺癌异种移植瘤弹性,对前列腺癌诊断具有重要的提示意义。
[Abstract]:Prostate cancer is a threat to life in elderly male malignant tumor in our country in recent years. By presenting the trend. Prostate cancer treatment protocol usually depends on whether the invasion and metastasis of the surrounding tissue. So the research aspects of early diagnosis and metastasis of prostate cancer is important parameter.FoxM1 and tumor disease; epithelial mesenchymal the transformation is an important link in the process of tumor metastasis. Shear wave elastography is an important method for identification of benign and malignant tumors. Based on the above research background, this research mainly through the detection of FoxM1 expression in prostate cancer tissues, to explore the mechanism and influence factors of the transfer of FoxM1 in prostate cancer EMT and human prostate cancer xenograft tumor of elastic the initial shear wave elastography, and analyze the pathological basis. Methods: 1FoxM1 expression in the identification of prostate cancer and detection of FoxM1 cells. The protein and gene expression in the subtype of prostate cancer, benign prostatic hyperplasia and prostate cancer cell lines, and analyze the relationship between FoxM1 expression and pathological Gleason score of prostate cancer. The detection of FoxM1 3 subtype FoxM1a, FoxM1b, FoxM1c in vitro in prostate cancer tissues and cell lines the expression of.2FoxM1 in prostate cancer cell lines EMT. The establishment of EMT prostate cancer cell model, detection of FoxM1 and EMT related molecules E-cadherin, vimentin, Slug expression, evaluation of cell movement ability; metformin and hypoxia treated prostate cancer cells, the expression of FoxM1 and EMT detection, assessment of cell proliferation, motility and migration of.3FoxM1 in animal experimental metastasis of prostate cancer. Construction of shRNA lentiviral vector carrying FoxM1 GFP, GFP FoxM1 shRNA and establish a stable expression in prostate cancer cell lines; nude mice transplantation, detection Tumor proliferation and metastasis of.4. shear wave elastography evaluation of transplanted tumor. The establishment of elastic prostate cancer xenograft model, ultrasound measurement of different growth time (6W, 8W, 10W) tumor size, shear wave elastography detection of tumors. Analysis of elastic fiber components of tumor tissue, and in vivo tumor type collagen the expression of alpha, -SMA detection of tumor tissue. FoxM1 knockdown xenograft model, the growth of 8W detected the target. Results: the expression of 1.FoxM1 in prostate cancer and benign prostatic hyperplasia tissue differences were expressed in prostate cancer cells, the Gleason expression was significantly higher than that of Gleason7 score more than 7 of the cancer tissues in (P0.05).FoxM1c subtype in prostate cancer cells and the expression of prostate cancer cell line EMT induced by high levels of.2.TGF-p vimentin and downregulation of E-cadherin, upregulation of Slug, cell transport capacity increase 844000 shift Strong, FoxM1 knockdown, EMT process was reversed. Metformin treated cells, the expression of endogenous FoxM1 decreased, decreased the proliferation and motility of cell migration; hypoxia cells increase the expression of FoxM1, promote prostate cancer cell EMT, cell movement and migration increased.3. constructed successfully lentiviral vector carrying GFP and FoxM1 shRNA, set up FoxM1 on.FoxM1 prostate cancer xenograft model of low knockdown tumor formation ability is weak, slow tumor growth, metastasis significantly reduced compared with the control group.4. were inoculated with 6w.8w.10w tumor volume were 832.58 + 14298mm31862.76 + 271.8mm3 and 2764.2 + 255.93mm3 (P0.05); tumor modulus were 22.11 + 2.45KPa, 44.89 + 3.02KPa and 57.41 + 3.98KPa (P0.05). The harder the tumor in the collagen fibers, collagen type I, and the expression of a-SMA is more.FoxM1 knockdown tumor than in the control group grew slowly, when inoculated with 8W tumor size respectively. 813.7686 + 111.1932mm3 and 1921.297 + 258.1513mm3 (P0.05); the young's modulus were 25.46 + 3.48KPa and 46.94 + 5.32KPa (P0.05, Fox) in M1 knockdown group collagen type I collagen composition is less and less, alpha ^ A expression less. Conclusion FoxM1 in prostate cancer was increased, and the score with prostate cancer is closely related to Gleason.FoxM1 plays an important role in prostate cancer EMT process, is expected to become a new target for treatment of prostate cancer. The shear wave elastography can evaluate the prostate cancer xenograft tumor elasticity, has important implications for diagnosis of prostate cancer.

【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R737.25

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