雄激素受体在良恶性前列腺组织中的表达及与细胞增殖凋亡的 …

一、材料与方法1.收集本院病理科 1977～ 1999年间手术切除及穿刺前列腺癌标本共 5 6例 ,其中切除标本 34例 ,穿刺标本 2 2例 ,所有标本均为 4%甲醛固定 ,石蜡包埋 ,连续切片 ,切片厚度 4μｍ。根据Ｇｌｅａｓｏｎ分级和临床医生分级标准将前列腺癌分为 3级 ,2～ 5分 (高分化 ) 8例 ,6～ 8分 (中分化 ) 32例 ,9～ 10分 (低分化 ) 16例。另选 18例良性前列腺增生症作对照组。2 .免疫组织化学采用超敏感ＳＰ法。雄激素受体 (ＡＲ)和Ｋｉ 6 7(ＭＩＢ 1)单克隆抗体为美国Ｍａｘｉｍ公司产品 ,分别用已知阳性乳腺癌和胃癌切片作ＡＲ和Ｋｉ 6 7阳…     

雄激素受体在人类前列腺癌发展中的作用研究

目的:研究雄激素受体在前列腺癌雄激素非依赖性生长中的作用。方法:通过连续传代培养,建立雄激素非依赖性前列腺癌细胞模型。用Western blot方法检测AR和PSA在上述过程中的表达情况。结果:连续传代培养后,LNCaP C-33细胞(传代次数少于33次)生物学行为发生改变。同C-33和C-51(传代次数介于33次和80次之间)细胞相比,C-81细胞(传代次数大于80次)表现出更强生长能力和更低雄激素依赖性。C-81细胞分泌高水平PSA,且DHT对其分泌作用的影响比C-33细胞小。3种LNCaP细胞总AR表达水平相同,但C-81表现为特征性AR-B表达丢失和AR-A表达增加。结论:前列腺癌进展是多因素作用的结果,其中包括AR亚型的差异性表达。     

激素受体及CgA表达与去势抵抗性前列腺癌的关系

目的探讨嗜铬粒蛋白A(chromogranin A, CgA)、AR、ER、PR表达与前列腺癌(prostate cancer, PCa)去势抵抗的关系。方法收集行TURP术有临床随访资料的63例PCa标本,采用免疫组化SP法检测CgA、AR、ER、PR在PCa组织中的表达情况,并分析去势抵抗与PCa各临床病理特征、CgA、AR、ER、PR的关系,以及CgA与PCa各临床病理特征、AR、ER、PR的关系。结果去势抵抗性PCa的发生与PCa PSA≥20 ng/mL、Gleason评分8～10、CgA阳性、AR阴性具有相关性,CgA阳性与PCa PSA≥20 ng/mL、Gleason评分8～10、AR阴性具有相关性。ER、PR表达与去势抵抗性PCa的发生及CgA表达无相关性。多因素Logistic回归分析结果显示Gleason评分是进展为去势抵抗性PCa的独立危险因素,AR是CgA的独立危险因素。结论 CgA表达与前列腺的恶性程度呈正相关,AR表达与前列腺的恶性程度呈负相关。Gleason评分、CgA及AR与PCa去势抵抗密切相关,监测CgA与AR的表达情况,结合Gleason评分对PCa的治疗方案选择和预后判断有一定的临床指导意义。     

45例人前列腺癌雄激素受体基因B~H外显子的突变研究

目的和方法：为探讨雄激素受体（ＡＲ）与前列腺癌（ＰＣ）发生发展及内分泌治疗不敏感的关系，我们首先建立了用ＰＣ的穿刺和石蜡切片组织检测ＡＲ基因突变的ＰＣＲ－ＳＳＣＰ（双链构象多态性）－双链ＤＮＡ循环测序法。并用上述方法检测了４５例国人前列腺癌组织ＡＲ基因Ｂ￣Ｈ我显子的突变。结果：在６例２的癌组织中证实有７个ＳＳＣ溘 变位片段（其中一患者有两个外显子异常）。这６例 ４例为低分化腺癌，其中２例已有远处转     

性激素对裸鼠移植性人前列腺癌（PC—3M）生长和雄激素受体水 …

用已烯雌酚（ＤＥＳ）和不同剂量的丙酸睾丸素（ＴＰ）治疗移植于裸鼠体内的雄激素非依赖性ＰＣ－３ｍ前列腺癌肿瘤模型，并以放射配体结合分析测定肿瘤的雄激素受体（ＡＲ）水平，治疗结束进测量瘤重和瘤体积。ＤＥＳ组的瘤重、瘤体积和ＡＲ水平与与照组的相比，差异不显著，说明ＤＥＳ对肿瘤的生长无明显抑制作用。与对照组的相比，低剂量ＴＰ（５０ｍｇ／ｋｇ）组的瘤重、瘤体积和ＡＲ水平显著增高（Ｐ〈０．０１）；而高剂量ＴＰ     

45例人前列腺癌雄激素受体基因B～H外显子的突变研究

目的和方法:为探讨雄激素受体(AR)与前列腺癌(PC)发生发展及内分泌治疗不敏感的关系,我们首先建立了用PC的穿刺和石蜡切片组织检测AR基因突变的PCR-SSCP(双链构象多态性)-双链DNA循环测序法.并用上述方法检测了45例国人前列腺癌组织(6例穿刺组织,39例石蜡切片)AR基因B～H外显子的基因突变.结果:在6例患者的癌组织中证实有7个SSCP泳动变位片段(其中一患者有两个外显子异常),这6例患者中4例为低分化腺癌,其中2例已有远处转移,而序列分析证实这2例转移患者SSCP异常的外显子H片段各存在一错义突变(Glu 872 Gln、Met 886 Ile).这两种AR的基因突变国内外均未见报道,为在PC中新发现的突变.结论:实验发现AR突变均发生在前列腺癌的中晚期,提示AR基因突变可能与PC的进展和转移有关.     

双标记方法检测骨肉瘤细胞增生和细胞凋亡

本研究应用ＡＰＡＡＰ和原位末端标记（ｉｎｓｉｔｕｅｎｄｌａｂｅｌｉｎｇ，ＩＳＥＬ）双标记方法，检测了８０例骨肉瘤细胞的增生和细胞凋亡，旨在建立一种同时显示细胞增生和细胞凋亡的方法，便于病理医生更好地观察肿瘤细胞增生和细胞凋亡的关系。     

双标忆方法检测骨肉瘤细胞增生和细胞凋亡

本研究应用ＡＰＡＡＰ和原位末端标记（ＩＳＥＬ）双标记方法，检测了８０例骨肉瘤细胞的增生和细胞凋亡，旨在建立一种同时显示细胞增生和细胞凋亡的方法，便于病理医生更好地观察肿瘤细胞增生和细胞凋亡的关系。     

利用RNA干涉技术研究雄激素受体在人前列腺癌细胞增殖中的作用

目的: 利用RNA干涉技术抑制雄激素受体(AR)的表达,研究AR在激素依赖性和激素非依赖性前列腺癌细胞增殖中的作用。方法: 设计、合成针对AR的4种不同的小干涉RNA(siRNA),连接到带有人H1启动子的腺病毒载体质粒pShuttle-H1-Ri中,构建成能产生AR-siRNA的质粒pShuttle-H1-Ri-AR,与能表达AR的质粒pcDNA-AR共转染HEK293细胞,Western blot 检测不同的siRNA对AR表达的抑制效率,选取抑制效率高的siRNA制备重组腺病毒,感染LNCap、C4-2B和CWR22Rv1 3种对雄激素有不同反应性的人前列腺癌细胞,采用Western blot 检测感染后细胞中AR的表达程度,并用MTT比色法测定细胞的增殖活性。结果: 4种siRNA都能抑制共转染AR的表达,用抑制效率高的siRNA制备重组腺病毒感染3种靶细胞后,均能特异性地抑制3种细胞中AR的表达,细胞的增殖活性也随之明显下降。结论: AR-siRNA通过抑制激素依赖性和激素非依赖性前列腺癌细胞中AR的表达,有效地抑制细胞的增殖,AR对维持激素依赖性和激素非依赖性前列腺癌细胞的增殖活性具有十分重要的作用,是治疗前列腺癌的重要靶分子。     

性激素对裸鼠移植性人前列腺癌（PC－3m）生长和雄激素受体水平的影响

用己烯雌酚（ＤＥＳ）和不同剂量的丙酸睾丸素（ＴＰ）治疗移植于裸鼠体内的雄激素非依赖性ＰＣ－３ｍ前列腺癌肿瘤模型，并以放射配体结合分析测定肿瘤的雄激素受体（ＡＲ）水平，治疗结束时测量瘤重和瘤体积。ＤＥＳ组的瘤重、瘤体积和ＡＲ水平与对照组的相比，差异不显著，说明ＤＥＳ对肿瘤的生长无明显抑制作用。与对照组的相比，低剂量ＴＰ（５０ｍｇ／ｋｇ）组的瘤重、瘤体积和ＡＲ水平显著增高（Ｐ＜０．０１）；而高剂量ＴＰ（４００ｍｇ／ｋｇ）组的瘤重、瘤体积和ＡＲ水平则显著降低（Ｐ＜０．０１）。这说明丙酸睾丸素对裸鼠体内ＰＣ－３ｍ前列腺癌的生长具有双相效应，即低剂量ＴＰ促进肿瘤的生长，高剂量ＴＰ抑制肿瘤生长。肿瘤的生长受到促进还是抑制可能与雄激素水平、ＡＲ水平的高低有关，低剂量、高剂量ＴＰ各自所具有的促瘤和抑瘤作用可能分别通过升高或降低肿瘤的ＡＲ水平这一环节来实现。     

Pleiotropic functional properties of androgen receptor mutants in prostate cancer

The androgen receptor (AR) signaling pathway plays an important role during the development of the normal prostate gland, but also during the progression of prostate cancer on androgen ablation therapy. Mutations in the AR gene emerge to keep active the AR signaling pathway and to support prostate cancer cells growth and survival despite the low levels of circulating androgens. Indeed, mutations affecting the ligand binding domain (LBD) of the AR have been shown to generate so-called "promiscuous" receptors that present widened ligand specificity and allow the stimulation of these receptors by a larger spectrum of endogenous hormones. Another class of mutations, arising in the amino-terminal domain (NTD) of the receptor, modulate AR interactions with coregulators involved in cell proliferation regulation. Besides characteristics of these well-known types of mutations, the properties of other classes of AR mutants recently described in prostate cancer are currently under investigation. Most interestingly, in addition to their potential role in the mechanisms which allow prostate cancer cells to escape androgen ablation therapy, data suggest that certain AR mutations are present early in the natural history of the disease and may play a role in many aspects of prostate cancer progression. Surprisingly, singular truncated AR devoid of their carboxy-terminal end (CTE) region seem to exert specific paracrine effects and to induce a clonal cooperation with neighboring prostate cancer cells, which may facilitate both the invasion and metastasis processes. In this article, we review the functional properties of different classes of AR mutants and their potential impact on the natural history of prostate cancer. Hum Mutat 0, 1-14, 2008. (c) 2008 Wiley-Liss, Inc.     

不同ER、PR状态乳腺癌中AR的表达及其意义

目的探讨AR在不同ER、PR状态乳腺癌中的表达及意义。方法采用免疫组化方法检测AR、ER、PR在173例乳腺癌中的表达,依据结果分组:(1)AR状态分组:AR阳性组和AR阴性组;(2)ER、PR状态分组:En组(ER、PR均阴性)、Ep组[ER和(或)PR阳性];(3)AR、ER、PR联合分组:En-AR+(En组且AR阳性)、En-AR-(En组且AR阴性)、Ep-AR+(Ep组且AR阳性)、Ep-AR-(Ep组且AR阴性),其中En-AR-又称为均阴性组,其他三组统称为部分或完全阳性组。不同分组方法比较与临床病理特征的关系。结果Ep组AR阳性率62.8%(54/86),En组AR阳性率37.9%(33/87),两组差异有显著性(P=0.001),AR阳性组体积小、核分裂少、组织学分级低(P0.05);En-AR-组表现为核分裂多、组织学分级高(P0.01),此外En组内AR阳性者核分裂少、组织学分级低(P0.05),Ep组内AR阳性者临床分期高(P=0.000),En-AR+、Ep-AR+、Ep-AR-比较均无差异。结论AR在不同激素状态乳腺癌中表达的意义不同,ER、PR均阴性乳腺癌表达AR者预后较好,ER、PR阳性乳腺癌表达AR者临床分期高。在选择针对性药物时应考虑到不同激素受体状态的组合。     

The androgen receptor co-activator CBP is up-regulated following androgen withdrawal and is highly expressed in advanced prostate cancer

The androgen receptor co-activator CREB (cAMP-response element binding protein)-binding protein (CBP) enhances androgen receptor activity after stimulation by androgenic hormones and androgen receptor antagonists. The aim of the present study was to investigate the regulation of CBP expression by steroid and peptide hormones in prostate cancer. For this purpose, LNCaP cells were treated with the synthetic androgen methyltrienolone (R1881), epidermal growth factor, insulin-like growth factor-I or interleukin-6 (IL-6). CBP protein and mRNA expression were studied by western blotting and real-time PCR, respectively. CBP expression was also investigated in tissue specimens obtained from 26 patients with therapy-resistant carcinoma of the prostate. In LNCaP cells, CBP protein was down-regulated by R1881 or IL-6. The non-steroidal anti-androgen bicalutamide antagonized the effects of R1881 and the Janus kinase inhibitor AG 490 reversed the effects of IL-6. In contrast, neither R1881 nor IL-6 caused any effect on CBP expression in the PC-3 cell line. In LNCaP cells, the inhibition of CBP expression by R1881 or IL-6 was also observed at the mRNA level. CBP protein was detected in all 26 specimens by immunohistochemistry. The results suggest that up-regulation of CBP during androgen ablation may be relevant to the failure of endocrine therapy in patients with prostate carcinoma.     

前列腺病变组织中细胞凋亡与bcl-2、bax、PCNA表达

目的　研究前列腺良、恶性病变组织中细胞增殖与凋亡及其与相关蛋白表达意义。方法　采用原位细胞凋亡标记技术 (TUNEL)及免疫组织化学ABC法对 36例前列腺癌 (PCa)、2 0例前列腺增生 (BPH)和 11例正常前列腺 (NP)组织石蜡切片bcl 2、bax、PCNA蛋白及细胞凋亡检测。结果　前列腺癌细胞凋亡指数 (AI)高于BPH和NP(P <0 0 1) ,bcl 2蛋白阳性表达者AI低于阴性者 (P <0 0 1) ,bax蛋白阳性表达者AI高于阴性者 (P <0 0 5 ) ;前列腺癌和BPH的bcl 2和PCNA蛋白阳性表达率高于NP(P <0 0 5 ) ,并随着肿瘤分级增高而增高 ;NP、BPH和前列腺癌的bax蛋白阳性表达率差异无显著性。前列腺癌细胞增殖指数 (PI)明显高于BPH(P <0 0 1) ,BPH细胞PI较NP明显增高 ,但细胞AI却显著下降。结论　细胞增殖与细胞凋亡的增加在PCa的发生和发展中起到了重要作用。在BPH的形成过程中前列腺组织细胞增殖增加而细胞凋亡减少 ,其中bcl 2和bax在细胞凋亡调节中起重要作用。     

Prognostic significance of neuroendocrine differentiation, proliferation activity and androgen receptor expression in prostate cancer

Androgen, acting via the androgen receptor (AR), is associated with the development and progression of prostate cancer. Anti-androgen therapy is widely used to manage prostate cancer. However, the conversion of the tumor from a hormone-sensitive to a hormone-insensitive status causes such therapy to fail. Several mechanisms have now been put forward for this conversion, including neuroendocrine (NE) differentiation of the tumor cells. In this study, we evaluated the prognostic significance of tumor-cell proliferation activity, NE differentiation and AR expression. Formalin-fixed, paraffin-embedded sections were prepared from 42 patients with adenocarcinoma of the prostate. Using antibodies to AR, the Ki-67 antigen (MIB-1), chromogranin A and synaptophysin, immunohistochemical expression of AR, tumor proliferation activity and NE differentiation were analyzed. Our study revealed that AR expression was significantly lower in adenocarcinoma (52.2 +/- 27.1%) than in non-tumorous prostate tissue (68.3 +/- 18.3%; P < 0.001). NE differentiation was found in 50% of the tumors, which was correlated with the Gleason score (P < 0.05). An univariate analysis revealed a significant correlation between progression-free survival with both AR expression (P < 0.01) and proliferation activity (P < 0.001). NE differentiation was not a prognostic factor in this study.     

Tyk2促进前列腺癌细胞系雄激素受体特异位点Tyr-267磷酸化

目的探讨JAK激酶家族成员Tyk2在雄激素受体(AR)磷酸化和前列腺癌细胞增殖中的作用。方法以LNCaP及LAPC-4细胞系为研究对象,在无雄激素条件下,用表皮生长因子(EGF)刺激,给予AIM-100/Baricitinib(INCB 028050)处理,免疫沉淀法和Western blot法检测AR磷酸化;将Tyk2 siRNA转染LNCaP及LAPC-4细胞系,用Western blot法检测沉默Tyk2对AR磷酸化的影响;用CCK-8试剂盒检测前列腺癌细胞增殖。结果 EGF诱导AR Tyr-267特异位点磷酸化并促进前列腺癌细胞增殖(P0.05);AIM-100抑制Ack1和Tyk2介导的LNCaP/LAPC-4细胞增殖(P0.05)和AR Tyr-267位点磷酸化;INCB抑制Tyk2磷酸化、AR Tyr-267位点磷酸化及LNCaP/LAPC-4细胞增殖(P0.05)。结论 Tyk2是EGF诱导AR Tyr-267特异位点磷酸化和前列腺癌细胞增殖的关键激酶,在前列腺癌发病过程中可能扮演重要角色。     

Amplification of the androgen receptor gene in bone metastases from hormone-refractory prostate cancer

The aim of this study was to examine the prevalence of androgen receptor (AR) amplification in metastases to bone and other sites in patients with hormone-refractory prostate cancer (HRPC) and to compare these findings with those in pretreatment primary tumour samples from the same patients. Tissue from 24 patients with HRPC was available for study, together with 13 primary tumour specimens. AR gene amplification and copy number for X-chromosome were assessed by fluorescence in situ hybridization (FISH) using a SpectrumOrange-labelled probe at locus Xq11-13 for the AR gene and a SpectrumGreen-labelled alpha-satellite probe for the X-chromosome (Vysis, UK, Ltd.). A minimum of 20 nuclei were scored in each of three tumour areas by two independent observers. Samples from 18/24 patients with HRPC (12 bone marrow biopsies, three local tumour recurrences, and three lymph nodes) and nine primary tumour specimens were adequate for FISH analysis. Results were expressed as a mean ratio of AR gene copy number : mean X-chromosome number, with a ratio of greater than 1.5 defined as amplification. AR gene amplification was seen in 9/18 (50%) cases of HRPC and in none of the primary (untreated) tumour specimens (p = 0.0048, Fisher's exact test). For the 12 bone marrow samples, AR gene amplification occurred in 5/12 (38%) cases. Elevated copy number for chromosome X occurred in 3/18 (17%) HRPC and 4/9 (44%) matched primary tumours. This study shows for the first time that AR gene amplification can be demonstrated by FISH in bone metastases from HRPC patients. Because bone marrow biopsies can be obtained from most patients with HRPC, the findings provide a rational basis for the routine selection of patients who may respond more favourably to second-line anti-androgen therapy.