雄激素受体在前列腺癌细胞系中作用的研究进展

前列腺癌是男性高发的恶性肿瘤之一,目前广泛运用的雄激素阻断治疗能抑制早期前列腺细胞生长,但最终多数肿瘤会复发并转变为侵袭能力更强的激素非依赖性前列腺癌.雄激素受体(AR)是前列腺上皮分化的调节因子,在调控前列腺癌进展的过程中发挥着重要作用.本文总结并比较了不同分化度的三种前列腺癌细胞LNcaP、PC3、DU145中AR...     

雄激素受体在雄激素非依赖性前列腺癌的表达

目的探讨雄激素受体(AR)在国人雄激素非依赖性前列腺癌(AIPC)的表达及其临床意义。方法采用免疫组化两步法,检测AR在15例AIPC和20例雄激素依赖性前列腺癌(ADPC)组织标本的表达水平。结果1例AIPC无AR表达,其余14例AIPC及全部ADPC均有AR高表达,AIPC染色的光密值为0.31±0.04较ADPC0.28±0.03升高。结论大多数国人AIPC均高表达AR,AR的异常表达可能在前列腺癌的进展中发挥重要的作用。     

雄激素受体在前列腺癌中表达和意义的探讨

目的 探讨雄激素受体（AR）在PCa中的表达和意义。方法 应用免疫组化Elivison法，检测45例PCa和10例前列腺增生组织标本中AR的表达，并探讨AR蛋白表达与PCa分期、分级、术前PSA、内分泌治疗效果及预后的关系。结果 AR在PCa和前列腺增生中阳性表达率分别为90％和93、3％，阳性表达率和表达程度无显著性差异（P〉0.05）；D期和A-C期PCa中阳性表达率分别为100％和87．5％，无显著差异（P〉0．05）．但D期PCa AR阳性表达程度显著增强（P〈0．05）；Gleason评分≤7和Gleason评分〉7PCa中阳性表达率分别为85.7％和100％，无显著差异（P〉0.05），但低分化PCa（Gleason评分〉7）AR阳性表达程度显著增强（P〈0．05）：PSA≤10ng／ml和PSA〉10ng／mlPEa中阳性表达率分别为80％和97.1％，阳性表达率和表达程度无显著性差异（P〉0．05）：在内分泌治疗有效和无效PCa中阳性表达率分别为87．5％和90、9％，阳性表达率和表达程度无显著性差异（P〉0．05）。结论 AR阳性表达程度与PCa分期、分级有关，对判断PCa生物学行为有参考价值。     

雄激素受体在晚期前列腺癌治疗中的作用

对34例晚期前列腺癌患者在接受抗雄激素治疗前作前列腺雄激素受体（AR）定量测定，以了解AR值在抗雌激素治疗中的作用。结果发现，前列腺癌细胞浆AR（cAR）和细胞核AR（nAR）的平均值分别为71．4±65．7fmol／mg和299．4±371．8fmol／mg；nAR值在100fmol／mg以上者为58．8％，多于cAR的26．5％。随访27例患者，在激素治疗有反应的17例中，nAR＞100fmol／mg者占76.5％，与cAR的17.6％有显著性差异。而在nAR≤100fmol／mg的患者中，60％于治疗8～24个月内反应性消火。认为nAR值的变化能较准确地预测前列腺癌患者对抗雄激素治疗的反应性。     

雄激素受体辅助因子与前列腺癌雄激素非依赖性的关系

前列腺癌是西方国家最常见的恶性肿瘤，是引起老年男性死亡的第二位原因。我国的前列腺发病率远低于欧美国家，但近年来有上升趋势。目前雄激素去势或者阻断仍然是晚期前列腺癌患者的首选治疗，然而，经这种治疗12～16个月后，前列腺癌由激素依赖性变成激素非依赖性，对抗雄激素治疗不再有效。尽管前列腺癌雄激素依赖性发生变化的机制目前尚不完全清楚，但大量的研究表明，前列腺癌雄激素非依赖性与雄激素受体(AR)的反常激活有关。     

前列腺组织与LNCaP细胞的雄激素受体亚型研究

目的 :研究雄激素受体 (AR)亚型在人前列腺良性增生与前列腺癌组织以及LNCaP细胞系中的表达 ,探讨AR亚型表达组织之间的差异。　方法 :应用氚标技术与聚丙烯酰胺凝胶等电聚焦电泳的方法 ,对相应组织的AR亚型进行分析。　结果 :4 1例AR阳性的前列腺增生组织、3例前列腺癌组织和LNCaP细胞中 ,发现有 3个AR亚型的表达 ,其等电点 (pI)分别在6 .5、6 .0和 5 .3。在前列腺增生标本中 ,15例 (36 .5 % )表达 pI6 .5、6 .0、5 .3亚型 ,10例 (2 4 .4 % )表达 pI6 .5、5 .3亚型 ,5例 (12 .2 % )表达 pI6 .5、6 .0亚型 ,4例 (9.8% )表达 pI6 .0、5 .3亚型 ,2例(4.9% )表达 pI6 .5亚型 ,2例 (4.9% )表达 pI6 .0亚型 ,3例 (7.3% )表达 pI5 .3亚型。在 3例前列腺癌组织中 ,1例表达pI 6 .5、6 .0、5 .3亚型 ,1例表达pI 6 .5、6 .0亚型 ,1例 3种亚型均不表达。LNCaP细胞表达pI 6 .5、6 .0、5 .3亚型。氚标的双氢睾酮 (DHT)与 3种受体亚型的结合 ,可以被非氚标的DHT、睾酮 (T)竞争抑制 ,而孕酮、雌二醇和己烯雌酚对此却无竞争抑制作用。　结论 :AR亚型的表达 ,在不同的病人与疾病和LNCaP前列腺癌细胞系之间存在差异 ,提示不同病人对前列腺癌的内分泌疗法有不同的反应性     

雄激素受体在晚期前列腺癌治疗中的作用

３４例晚期前列腺癌病人在接受抗雄激素治疗前作前列腺雄激素受体（ＡＲ）定量测定，以了解ＡＲ值在抗雄激素治疗中的作用。结果发现：前列腺癌细胞浆ＡＲ（ｃＡＲ）和细胞核ＡＲ（ｎＡＲ）的平均值（ｆｍｏｌ／ｍｇＤＮＡ）分别为７１．４±６５．７和２９９．４±３７１．８；ｎＡＲ值在１００以上的例数为５８．８％，多于ｃＡＲ的２６．５％。随访２７例病人，对激素治疗有反应的１７例中，ｎＡＲ＞１００的例数占７６．５％，与ｃＡＲ的１７．６％差异有显著性。而在ｎＡＲ≤１００ｆｍｏｌ的病人中，６０％于治疗８～２４个月内反应性消失。认为ｎＡＲ值的变化能较准确地预测前列腺癌病人对抗雄激素治疗的反应性。     

雄激素受体与激素非依赖性前列腺癌

晚期前列腺癌可发生激素非依赖，对抗雄激素治疗不再有效。前列腺癌对雄激素依赖性发生变化的机制目前尚未阐明，近年来的研究显示前列腺癌雄激素非依赖性与雄激素受体(AR)的反常激活有关，与雄激素受体(AR)表达增加和突变、AR调节因子表达改变有关，细胞因子IL-6、IL-4，糖原合成激酶-3β参与激素非依赖性前列腺癌的AR激活。本文就这方面的研究进展作一综述。     

雄激素受体与前列腺癌关系的研究进展

目前前列腺癌还缺乏能较好地预示其预后和内分泌的治疗反应的指标。雄激素受体在这方面的作用已受到人们的重视，为此本文对近年来雄激素受体与前列腺癌发生发主内分泌治疗之间关系的研究进行综述。雄     

雄激素受体与激素非依赖性前列腺癌

晚期前列腺癌可发生激素非依赖,对抗雄激素治疗不再有效。前列腺癌对雄激素依赖性发生变化的机制目前尚未阐明,近年来的研究显示前列腺癌雄激素非依赖性与雄激素受体(AR)的反常激活有关,与雄激素受体(AR)表达增加和突变、AR调节因子表达改变有关,细胞因子IL-6、IL-4,糖原合成激酶-3β参与激素非依赖性前列腺癌的AR激活。本文就这方面的研究进展作一综述。     

Effect of the neuropeptides bombesin and calcitonin on the growth of prostate cell lines, PC-3, DU 145, and LNCaP

Neuroendocrine cells are present in normal and tumoral prostate tissue, the neuropeptides secreted by this cells have a biological functions that have not been fully elucidated. The presence of neuroendocrine cells in prostatic carcinoma have been shown to increase tumor progression. We characterized the in vitro proliferative influence of bombesin and calcitonin in androgen-insensitive, PC-3 and DU-145, and androgen-sensitive, LNCaP, cell lines of human prostate cancers. The influence of these neuropeptides on proliferation were assessed using the colorimetric XTT assay and by cells counts with a hemocytometer. The growth of PC-3 and DU-145 cell lines is stimulated by bombesin and calcitonin but exerted any stimulatory effect on the proliferation of the LNCaP cell line. This indicate that bombesin and calcitonin can modulate proliferation of androgen-insensitive human prostate cell lines "in vitro" and may be potential paracrine growth promoters in stablished androgen irresponsive human prostatic carcinoma cells.     

Resveratrol induces apoptosis in LNCaP cells and requires hydroxyl groups to decrease viability in LNCaP and DU 145 cells

BACKGROUND: This study was conducted to determine the effects of resveratrol on prostate cancer cell viability through apoptosis induction and the significance of the three hydroxyl groups on resveratrol to the measured effect. METHODS: Hormone-sensitive LNCaP cells and hormone-insensitive DU 145 cells were treated with resveratrol, tri-methoxy-resveratrol, or diethylstilbestrol (DES; the positive control for toxicity and apoptosis). Cell viability was determined by using an MTS assay. Apoptosis was determined by the appearance of apoptotic morphology, annexin V-FITC-positive intact cells, and caspase activation. RESULTS: Resveratrol and DES decreased viability in LNCaP cells, but only resveratrol-treated cells expressed apoptotic morphology, annexin V-FITC-positive cells, and caspase activation. Tri-methoxy-resveratrol had no effect on DU 145 cell-viability and was less toxic to LNCaP cells than resveratrol. CONCLUSION: Resveratrol was toxic to cells regardless of whether the cells were hormone-responsive or -unresponsive. This finding suggests that the cell's hormone responsive status is not an important determinant of the response to resveratrol. Furthermore, the hydroxyl-groups on resveratrol are required for cell toxicity. Finally resveratrol but not DES induced caspase-mediated apoptosis.     

雄激素受体在乳腺癌细胞中的表达及其对细胞增殖的影响

Objective To evaluate the expression of androgen receptor (AR) in the breast cancer cell lines and its effect on proliferation of breast cancer cells. Methods The estrogen receptor (ER) -positive MCF-7 and ER-negative MDA-MB-453 cells were involved in this study and cultured in vitro. The expression of AR was detected by using Western blotting. Cell proliferation was determined by methyl thiazol tetrazolium (MTT) assay after the treatment with different concentrations of dihydrotestosterone (DHT) ( 1 x 10 -7, 1 x 10- 8, 1 x 10 -9 mol/L) for 48, 96 and 144 h respectively. Cell cycle was analyzed by flow cytometry following culture for 72 h. Results DHT increased the AR expression in the two breast cancer cell lines. AR pathway could inhibit proliferation of MCF-7 and MDA-MB-453 cells. There was no significant difference in absorbance values among three treatment groups at different time points (P ＞ 0. 05). Cell cycle analysis revealed that the proportion of cells at G1 phase was increased, and that at S phase decreased. Conclusion AR pathway may inhibit proliferation of ER-negative MDA-MB-453 breast cells as well as ER-positive MCF-7 cells, by suppressing the process of G1 to S phase progression.     

雄激素受体在乳腺癌细胞中的表达及其对细胞增殖的影响

目的 检测雄激素受体(AR)在乳腺癌细胞中的表达,并观察雄激素刺激对乳腺癌细胞增殖的影响.方法 选择雌激素受体(ER)阳性的MCF-7和ER阴性的MDA-MB-453乳腺癌细胞,体外培养,Western blot技术检测两乳腺癌细胞株中AR蛋白的表达,MTT法检测用1×10-7、1×10-8和1×10-9 mol/L不同浓度的雄激素二氢睾酮(DHT)分别干预48、96、144 h后的细胞增殖,并应用流式细胞术检测DHT刺激乳腺癌细胞72 h后细胞周期的变化.结果 两个乳腺癌细胞株经DHT作用后AR蛋白表达均增多,DHT通过AR抑制MCF-7和MDA-MB-453两个乳腺癌细胞株的生长,各时间段不同浓度组比较A值差异无统计学意义(P＞0.05),细胞周期结果显示G1期细胞比例增高,S期细胞比例降低.结论 雄激素受体途径对ER阳性的MCF-7和ER阴性的MDA-MB-453乳腺癌细胞均有抑制生长作用,可能通过抑制细胞由G1期到S期转化来实现的.

Abstract：

Objective To evaluate the expression of androgen receptor (AR) in the breast cancer cell lines and its effect on proliferation of breast cancer cells. Methods The estrogen receptor (ER) -positive MCF-7 and ER-negative MDA-MB-453 cells were involved in this study and cultured in vitro. The expression of AR was detected by using Western blotting. Cell proliferation was determined by methyl thiazol tetrazolium (MTT) assay after the treatment with different concentrations of dihydrotestosterone (DHT) ( 1 x 10 -7, 1 x 10- 8, 1 x 10 -9 mol/L) for 48, 96 and 144 h respectively. Cell cycle was analyzed by flow cytometry following culture for 72 h. Results DHT increased the AR expression in the two breast cancer cell lines. AR pathway could inhibit proliferation of MCF-7 and MDA-MB-453 cells. There was no significant difference in absorbance values among three treatment groups at different time points (P ＞ 0. 05). Cell cycle analysis revealed that the proportion of cells at G1 phase was increased, and that at S phase decreased. Conclusion AR pathway may inhibit proliferation of ER-negative MDA-MB-453 breast cells as well as ER-positive MCF-7 cells, by suppressing the process of G1 to S phase progression.     

雄激素受体在乳腺癌细胞中的表达及其对细胞增殖的影响

Objective To evaluate the expression of androgen receptor (AR) in the breast cancer cell lines and its effect on proliferation of breast cancer cells. Methods The estrogen receptor (ER) -positive MCF-7 and ER-negative MDA-MB-453 cells were involved in this study and cultured in vitro. The expression of AR was detected by using Western blotting. Cell proliferation was determined by methyl thiazol tetrazolium (MTT) assay after the treatment with different concentrations of dihydrotestosterone (DHT) ( 1 x 10 -7, 1 x 10- 8, 1 x 10 -9 mol/L) for 48, 96 and 144 h respectively. Cell cycle was analyzed by flow cytometry following culture for 72 h. Results DHT increased the AR expression in the two breast cancer cell lines. AR pathway could inhibit proliferation of MCF-7 and MDA-MB-453 cells. There was no significant difference in absorbance values among three treatment groups at different time points (P ＞ 0. 05). Cell cycle analysis revealed that the proportion of cells at G1 phase was increased, and that at S phase decreased. Conclusion AR pathway may inhibit proliferation of ER-negative MDA-MB-453 breast cells as well as ER-positive MCF-7 cells, by suppressing the process of G1 to S phase progression.     

Sorafenib对前列腺癌DU145细胞的抑制作用的研究

目的 观察索拉非尼(Sorafenib)对雄激素非依赖性前列腺癌DU145细胞的抑制作用.方法 用不同浓度Sorafenib处理前列腺癌DU145细胞24、48和72 h后,MTT法检测Sorafenib对DU145细胞的抑制作用,流式细胞仪检测细胞凋亡变化,Western blot检测不同浓度Sorafenib处理72 h后DU145细胞内ERK和Bcl-2的表达.结果 Sorafenib能显著抑制DU145细胞的体外生长,呈时间与剂量依赖性.DU145细胞凋亡率随着Sorafenib剂量的增加而增大,具有良好的量效关系(P<0.01);Sorafenib处理DU145细胞72 h后,ERK和Bcl-2蛋白的表达明显下调(P<0.01).结论 Sorafenib抑制DU145细胞增殖、诱导细胞凋亡,可显著抑制雄激素非依赖性前列腺癌细胞的体外生长.     

Estramustine-induced mitotic arrest in two human prostatic carcinoma cell lines DU 145 and PC-3

In growth proliferation experiments on two human prostatic carcinoma cell lines, DU 145 cells were found to be more sensitive to the cytotoxic effect of estramustine and nor-nitrogen mustard than PC-3 cells. Estramustine was, however, much more cytotoxic in both cell lines than nor-nitrogen mustard. Cytogenetic experiments revealed that estramustine produced a drastic increase of the mitotic index in both these cell lines. This increase could be accounted for by the arrest of cells in their first treatment-metaphase. The arrested metaphases exhibited all the characteristics commonly found for stathmokinetic agents such as colchicine and vinca-analogues. No mitotic arrest was found for nor-nitrogen mustard but chromosomal aberrations were found at toxic concentrations. Estradiol exhibited minimal toxicity and caused no mitotic arrest in these cell lines. The mitotic arrest induced by estramustine was found to be reversible on removal of the drug.     

Inhibition of cell signaling by the combi-nitrosourea FD137 in the androgen independent DU145 prostate cancer cell line

BACKGROUND: FD137, a nitrosourea appended to a quinazoline ring, was designed to simultaneously block epidermal growth factor receptor (EGFR)-mediated signaling and damage genomic DNA in refractory EGF-dependent prostate tumors. METHODS: The mixed inhibition of cell signaling and DNA damage by FD137 were determined by Western blotting, RT-PCR, flow cytometry, sulforhodamine B (SRB), and comet assay. RESULTS: FD137 and its metabolite FD110 induced a dose-dependent increase in inhibition of EGF-stimulated EGFR autophosphorylation and this translated into blockade of c-fos gene expression in DU145 cells. FD137 induced significant levels of DNA damage and showed 150-fold greater anti-proliferative activity than BCNU, a classical nitrosourea. In contrast to BCNU, complete inhibition of EGF-induced cell transition to S-phase was observed at concentrations of FD137 as low as 3 microM. CONCLUSION: FD137 could not only damage DNA, but also significantly block downstream EGFR-mediated signaling. The superior activity of FD137 may be imputable to the combined effect of its mixed EGFR/DNA targeting properties. This novel strategy may well represent a new approach to target nitrosoureas to EGFR-overexpressing carcinomas of the prostate.     

The expression of estramustine-binding protein in the human prostatic cancer cell line DU 145 is not androgen dependent.

Estramustine-binding protein (EMBP) constitutes one of the major proteins in the prostatic gland, it binds estramustine and estromustine, the active metabolites of estramustine phosphate (Estracyt). Previous studies in rats have indicated that the expression of EMBP is androgen dependent, with diminishing quantities following castration and estrogen treatment as well as restored pre-castration production upon administration of androgens. In this study, we have used the human prostatic cancer cell line DU 145 transplanted in female and male nude mice. This cell line, which is sex hormone independent, gave rise to subcutaneous tumors in the rats with no difference in growth characteristics between the males and females. The expression of EMBP was analysed by radioimmunoassay, immunohistochemical and Western blot techniques. No difference was seen between the two sexes with respect to EMBP content, demonstrating that the expression of EMBP, in contrast to that reported for the normal prostate, is neither androgen- nor estrogen-dependent in tumor tissues.