中药百合
PCa prostate cancer 前列腺癌
PCR Polymera chain reaction 聚合酶链式反应PI3K Phosphatidyl inositol-3 kina 磷脂酸肌醇-3激酶PSA prostate-specific antigen 前列腺特异性抗原RNA ribonucleic acid 核糖核酸
rpm rounds per minute 转/min
RT-PCR rever transcription PCR 逆转录PCR
s cond 秒
SBP spermine-binding protein 精胺结合蛋白SHH Sonic hedgehog 脑音猬因子SLUG zinc-finger transcriptional factor 锌指转录因子
SP-B pro-surfactant protein-B 表面活性蛋白B SP-C 表面活性蛋白C 表面活性蛋白C
STAT Signal transducers and activators of
品读国学经典
transcription
信号传导及转录激
活因子
TEMED tetramethylethylenediamine 四甲基乙二胺Tris trihydroxymethyl aminomethane 三羟甲基氨基甲烷
FOXA1在前列腺癌中的表达及临床相关性研究
硕士研究生：冯伟
导师：张红宾教授
第四军医大学西京医院泌尿外科，西安，710032
联合培养单位：兰州军区总医院泌尿外科，兰州，730050
中文摘要
前列腺癌(prostate cancer，PCa)是男性生殖系最常见的恶性肿瘤，不同地区和不同人种的发病率具有明显差异，并且随年龄增长而增长。欧美地区发病率较高，在男性癌症死亡原因中PCa位居第二，仅次于肺癌[1, 2]，在我国前列腺癌发病率也呈增高趋势，有统计显示城市与农村发病率差异较大。目前
研究发现，前列腺癌的发病和许多癌基因及抑癌基因NKX3.1、Ras、Myc、Sis、Fos、her1（erbB1）、her2（erbB2）、P53有关，也涉及到许多信号通路的参加，如PI3K-Akt-mTOR、Ras-Raf-Mek-MAPK、Stat等。Forkhead-boxA1(FOXA1)基因亦为hepatoeyte nuclear factor 3a(HNF3a)基因，是转录因子FOX家族中的一员[3]，它在上皮细胞分化的过程起着至关重要的作用，现有研究其在人体许多组织如乳腺、肺、食管、肝、胰腺、膀胱和前列腺等部位中可见表达[4-6]。FOXA1可通过与基因启动子结合，起到调节细胞的信号与周期的作用[7, 8]。作为性激素相关性肿瘤，前列腺癌的发生发展与雄激素密切相关，而FOXA1作为公认的雄激素受体相关基因，在前列腺癌中的变化和作用必然引起关注，但有关FOXA1与前列腺癌的关系国内尚未见研究报道。为探索FOXA1与前列腺癌的相关性，本课题将石蜡组织标本和新鲜组织标本结合，应用免疫组织化学染色、Western-blot、Real-Time PCR三种实验方法，从蛋白和核酸两个层面，观察FOXA1在前列腺癌和前列腺增生（Benign prostatic hyperplasia，BPH）组织中的定位和表达
有效课堂情况，并结合患者临床资料，分析FOXA1阳性表达率与患者年龄、肿瘤TNM分期、Gleason分级之间的关系，探讨FOXA1在前列腺癌中的致癌机制。
具体实验设计如下：
目的：探讨FOXA1表达与前列腺癌患者年龄、肿瘤TNM分期、Gleason分级之间的关系。
高考压轴题
方法：收集前列腺癌根治术后和前列腺穿刺活检石蜡组织35例、前列腺增生电切除术后石蜡组织21例，应用免疫组化法检测FOXA1蛋白的定位和表达情况；收集前列腺癌根治术后冰冻组织21例，前列腺增生电切除术后冰冻组织21例，应用Western-blot和Real-Time PCR方法检测FOXA1蛋白和核酸的表达情况；应用统计软件分析FOXA1在mRNA转录和总蛋白表达过程中的相互关系。
结果：
1.免疫组化染色显示，FOXA1在前列腺癌和前列腺增生组织中都有阳性表达，表达位置在前列腺腺上皮细胞细胞核。前列腺癌组织中FOXA1表达率明显高于前列腺增生组织（P<0.001）。
2.免疫组化染色中，通过半定量分析，FOXA1的表达阳性率与患者年龄无明显相关性，但与Gleason评分（P=0.027）和肿瘤TNM分期（P=0.003）呈正相关。
3.Western-blot检测显示，在裂解细胞总蛋白中，FOXA1蛋白在前列腺癌组织中的表达明显高于前列腺增生组织（P<0.001）。
4.Real-Time PCR检测显示，FOXA1核酸转录mRNA在前列腺癌组织中明显高于前列腺增生组织（P<0.001）。
有效工作时间
结论：任正非文章
1. FOXA1在前列腺癌的核酸转录mRNA和蛋白水平均为高表达，提示FOXA1与前列腺癌高度相关。
2.前列腺癌的Gleason评分和TNM分期越高，FOXA1的表达越强，提示FOXA1阳性表达与前列腺癌的恶性程度相关。
3.前列腺癌中核酸转录mRNA与蛋白表达呈正相关，提示FOXA1参与调控前列腺癌的发生和发展过程。
关键词：前列腺癌，FOXA1，肿瘤，基因游戏人的家
FOXA1 expression and clinical correlation
in prostate cancer
Candidate for master:Feng Wei
Supervisor:Zhang Hongbin
Department of Urology，Xijing Hospital，Fourth Military Medical University
Xi’an Shannxi 710032，China
Department of urinary surgery center, Lanzhou military region general hospital
Lanzhou Gansu 730050，China
Abstract
Prostate cancer (PCa) is the most common malignant tumor in male reproductive system. The incidence of this dia increas along with the age growth and has obvious regional and ethnic difference, which is much higher in Europe and America. PCa is the cond leading cau of death of cancer in men, cond only to lung cancer [1, 2]. In our country, the incidence of PCa also exhibits a trend of increa and there are statistics show that there is a big difference between the incidence in urban and rural. Prent studies have found that the pathogenesis of PCa is related with many cancer genes such as NKX3.1, Ras, Myc, Sis, Fos, her1 (erbB1), her2 (erbB2) and also involves many signaling pathways, such as PI3K-Akt-mTOR, Ras-Raf-Mek-MAPK, Stat, etc. Forkhead - boxA1 (FOXA1) gene,which is also called hepatoeyte nuclear factor 3 a (HNF3a) gene, is a member of transcription factors FOX family[3]and plays an important role in the differentiation of epithelial cell. Prent studies have established that FOXA1 is expresd in many human organs, such as breast, lung, esophagus, liver, pancreas, bladder, prostate茵陈汤
and so on[4-6] and can combine to multiple gene promoters to regulate cell signaling and cell cycle [7, 8]. But the relationship between FOXA1 and PCa has not been reported in domestic rearches and only veral studies involve this relationship in foreign rearches. To explore the correlation between FOXA1 and PCa, this study combines paraffin specimens and fresh tissue specimens and u immunohistochemical staining, Western blot, Real-Time PCR to obrve the location and expression of FOXA1 in PCa and benign prostatic hyperplasia (BPH) from the level of protein and nucleic acid. Combined with clinical data, we also analyzed the relationship between FOXA1 positive expression rate and patients' age, the tumor TNM staging and Gleason grading.
The design of experiment is as follows:
Objective: Explore the relationship between FOXA1 positive expression rate and patients' age, the tumor TNM staging and Gleason grading.
Methods: Collect 35 paraffin specimens of prostate cancer radical surgery and prostate biopsy and 21 paraffin specimens after electric rection of prostatic hyperplasia and u immunohistochemical staining to detect the localization and expression FOXA1 protein; Collect 21 frozen tissues after prostate cancer radical surgery and 21 frozen tissues after electric rection of prostatic hyperplasia
and u Western blot and Real Time-PCR to detect the expression of FOXA1 protein and nucleic acid; U statistical software to analysis mutual relations between FOXA1 mRNA transcription and total protein expression.
Results:
1. The result of immunohistochemical staining showed that FOXA1 was both expresd in prostate cancer and prostate hyperplasia tissues and was located in prostate gland epithelial cell nuclei. The expression of FOXA1 in prostate cancer tissue is significantly higher than in the prostate hyperplasia tissue (P<0.001).
2. Through a quantitative analysis, the result of immunohistochemical staining showed that the expression of FOXA1 had no obvious correlation with the patients' age, but was positively correlated with Gleason score (P=0.027) and tumor TNM stage (P=0.003).
3. The result of Western-blot test showed that within the cell total protein, the expression
