孕激素受体及B亚型在子宫内膜腺癌的表达及意义

目的 研究子宫内膜癌组织中孕激素受体及B亚型的分布及表达,探讨其临床意义。方法选取30例子宫内膜癌手术标本和10例正常增生期子宫内膜标本,石蜡切片,采用免疫组化方法,测定组织孕激素受体亚型的分布及表达。结果孕激素受体及B亚型均为核受体。PR在子宫内膜癌组织中表达的阳性率为93.33％,对照组的阳性率为100.00％,两者比较差异无显著性(P>0.05);PRB在子宫内膜癌组织中表达的阳性率为50.00％,与对照组比较差异有显著性(P<0.05)。PR的阳性率表达与分化程度无相关性,肿瘤不同分化程度的PRB阳性表达率有显著差异,分化高的:PRB阳性率高,分化低的PRB阳性率低。PR及PRB的阳性表达在临床分期I～Ⅲ各组间差异均无显著性(P值均>0.05)。结论hPR与hPRB为核受体,在子宫内膜腺癌组织中的阳性表达有所不同,PRB的阳性表达与病理分级有一定的关系,故测定PR、PRB在内膜癌的表达对指导其内分泌治疗、预测预后有一定的意义。     

人类胰岛素样生长因子-Ⅰ调控子宫内膜癌细胞系孕激素受体亚型表达的研究

目的探讨不同剂量人类胰岛素样生长因子-Ⅰ(IGF-Ⅰ)对子宫内膜癌细胞系中孕激素受体亚型表达的动态变化的调控.方法体外培养子宫内膜癌细胞系HEC-IB,以乳腺癌细胞系MCF-7为对照,采用蛋白印迹(Westernblot)法观察不同剂量IGF-Ⅰ对两种孕激素受体亚型表达的动态变化的调控.结果 (1)HEC-IB细胞中,IGF-Ⅰ 10 ng/ml作用24h,人孕激素受体B亚型(hPRB)表达有明显上调作用,随着剂量的增加、作用时间的延长,hPRB表达逐渐下调,IGF-Ⅰ 20 ng/ml作用72h及40 ng/ml作用48 h时,下调最明显.人孕激素受体A亚型(hPRA)表达的变化趋势与hPRB大致相同,IGF-Ⅰ为20 ng/ml作用48h时下调至最低.(2)MCF-7细胞中,IGF-Ⅰ 10、40 ng/ml对hPRA、hPRB表达均有上调的作用,在第24、48、72h时较为明显.IGF-Ⅰ 20 ng/ml作用24 h对hPRB表达有上调的作用,但在48、72h出现明显的下调;而对hPRA表达均为下调作用.结论 (1)IGF-Ⅰ对人孕激素受体亚型的调控具有细胞特异性、以及时间和剂量的依赖性.(2)HEC-IB细胞中,IGF-Ⅰ 10 ng/ml作用24 h对hPRA、hPRB表达有上调作用,随着剂量的增加、作用时间的延长,hPRA、hPRB表达渐下调.     

人类胰岛素样生长因子—I调控子宫内膜癌细胞系孕激素受体亚型表达的研究

目的 探讨不同剂量人类胰岛素样生长因子－I（IGF－I）对子宫内膜癌细胞系中孕激素受体亚型表达的动态变化的调控。方法 体外培养子宫内膜癌细胞系HEC－IB,以乳腺癌细胞系MCF－7为对照,采用蛋白印迹（Western blot)法观察不同剂量IGF－I对两种孕激素受体亚型表达的动态变化的调控。结果 （1）HEC－IB细胞中,IGF－I 10ng/ml作用24h,人孕激素受体B亚型（hPRB)表达有明显上调作用,随着剂量的增加、作用时间的延长,pPRB表达逐渐下调,IGF－I 20ng/ml作用72h及40ng/ml作用48h时,下调最明显。人孕激素受体A亚型（hPRA)表达的变化趋势与hPRB大致相同,IGF－I为20ng/ml作用48h时下调至最低。（2）MCF－7细胞中,IGF－I 10、40ng/ml对hPRA、hPRB表达均有上调的作用,在第24、48、72h时较为明显。IGF－I 20ng/ml作用24h对hPRB表达有上调的作用,但在48、72h出现明显的下调;而对hPRA表达均为下调作用。结果 （1）IGF－I对人孕激素受体亚型的调控具有细胞特异性、以及时间和剂量的依赖性。（2）HEC－IB细胞中,IGF－I 10ng/ml作用24h对hPRA、hPRB表达有上调作用,随着剂量的增加、作用时间的延长,hPRA、hPRB表达渐下调。     

层粘连蛋白受体基因表达与子宫内膜癌的关系

目的 探讨67ku层粘连蛋白受体(67LR)基因表达与子宫内膜癌的关系。方法 应用免疫组化SABC法检测1990～2001年收治的44例子宫内膜癌、15例子宫内膜增殖症、47例正常子宫内膜组织中67LR的表达;采用荧光定量逆转录聚合酶链反应(FQ-RT-PCR)方法定量检测2000～2001年收治的24例子宫内膜癌、16例子宫内膜增殖症、29例正常子宫内膜新鲜组织67LRmRNA水平。结果 67LR在子宫内膜癌、子宫内膜增殖症的表达较正常子宫内膜均明显增高(P<0.001),但前两组的表达差异无显著性(P>0.05)。高、中分化子宫内膜腺癌67LR表达较低分化者为高(P<0.05);但高、中、低分化组病例67LR mRNA水平差异无显著性(P>0.05)。67LR mRNA水平与免疫组化蛋白表达无明显相关(P>0.05)。结论 67LR蛋白过度表达可能与子宫内膜癌的发生、发展有关。67LR mRNA水平与蛋白表达水平无相关。     

雌激素对子宫内膜癌孕激素受体亚型调控的研究

目的 研究不同剂量雌激素(雌二醇,E_2)对子宫内膜癌细胞系中孕激素受体亚型在蛋白水平的调控,探讨雌激素、孕激素受体亚型与子宫内膜癌的关系。方法 体外培养子宫内膜癌细胞系HEC-IB,分别用含5nm、10nm、20nm雌二醇的培养液分别作用于细胞24、48、72h,以不含雌二醇的培养液培养细胞为自身对照。通过Western blot方法观察不同剂量雌激素对孕激素受体亚型A、B的蛋白水平调控的动态变化,并以乳腺癌细胞系MCF-7为对照。结果 ①HEC-IB细胞分别经5nm、10nm、20nm的E_2作用24、48、72h后,与未加药的细胞相比,不同浓度、时间下E_2对hPRB调控作用均不明显;而对hPRA的作用在E_2 5um时下调,且随时间延长,下调作用越加明显;在10nm的第24、48h下降,其后渐上升,而在E_2 20nm作用72h时基本恢复加药前的水平。②MCF-7细胞分别经E_2 5nm、10nm、20nm作用24、48、72h后,与未加药的细胞相比较,各种浓度及时间下,E_2对hPRA、B有上调作用的趋势;其中10nm的E_2上调作用较为明显,各种浓度的E_2在第48h的上调作用更为显著。结论 E_2对hPR亚型的调控具有细胞特异性、时间及剂量的依赖性。HEC-IB细胞中,各种浓度、时间下E_2对hPRB调控作用均不明显;而对hPRA的调控作用具有下调的趋势。     

子宫内膜癌中孕激素受体亚型表达及其甲基化状态的研究

目的:研究子宫内膜癌中孕激素受体亚型PRA、PRB表达与其启动子区甲基化状态的关系,探讨子宫内膜癌中孕激素受体亚型表达下调的机制。方法:(1)应用Real-Time PCR法检测子宫内膜癌及正常组织中孕激素受体亚型PRA、PRB mRNA的表达及去甲基化试剂5-Aza-CdR处理Ishikawa细胞前后孕激素两种受体亚型及DNA甲基化转移酶mRNA的表达;(2)应用甲基化特异性PCR(MS-PCR)检测子宫内膜癌细胞系Ishikawa中PRA、PRB的甲基化状态。结果:与正常子宫内膜组织比较,子宫内膜癌组织中,总PR表达显著降低,与病理类型相关;PRB表达显著下降,与其临床特征无关。子宫内膜癌细胞系Ishikawa中,加入5-Aza-CdR2.5μmol/L48h后,孕激素受体亚型PRA、PRB甲基化条带均消失;孕激素受体亚型PRA、PRB mRNA表达水平增加;同时Ishikawa细胞中DNA甲基转移酶DNMT1、DNMT3B表达下调。结论:子宫内膜癌组织中,孕激素受体亚型mRNA表达下调可能与子宫内膜癌的发生相关;孕激素受体亚型表达下调与其启动子区甲基化状态相关;去甲基化试剂5-Aza-CdR可逆转基因启动子区的甲基化状态,恢复PR基因的表达。     

子宫内膜癌雄激素受体表达及临床意义

目的:探讨子宫内膜癌组织中的雄激素受体(AR)的表达及其与临床病理特征和雌激素受体(ER)、孕激素受体(PR)表达的关系。方法:应用免疫组织化学SP法检测41例正常子宫内膜、18例不典型增生及116例子宫内膜癌组织中AR、ER、PR的表达。结果:①子宫内膜细胞普遍存在AR的表达,在正常子宫内膜、不典型增生子宫内膜、子宫内膜癌组织中阳性表达率逐渐增高,但差异无统计学意义(P=0.424)。②AR在子宫内膜癌中的表达随患者FIGO分期、组织病理分级的升高而下降(P=0.011;P=0.047),而与患者发病年龄、是否绝经、组织学类型、淋巴结有无转移、肌层有无浸润无明显关系(P>0.05)。③AR的表达与ER、PR的表达呈正相关(r=0.293,P=0.001;r=0.275,P=0.003)。结论:AR在子宫内膜癌的发生、发展中可能起重要作用,AR阳性表达者的生物学行为较好。     

抗苗勒管激素及其Ⅱ型受体在子宫内膜样腺癌中的表达

目的:观察抗苗勒管激素(AMH)及其Ⅱ型受体(AMHR-Ⅱ)在子宫内膜样腺癌组织中的表达及分布,探讨二者在子宫内膜样腺癌发生发展中的作用。方法:采用免疫组织化学方法对39例不同分化级别(高分化腺癌16例、中分化腺癌13例、低分化腺癌10例)的子宫内膜样腺癌组织进行AMH及AMHR-Ⅱ的检测,并对其阳性表达进行比较。结果:在不同分化级别的子宫内膜样腺癌组织中均有AMH及AMHR-Ⅱ的阳性表达,免疫反应阳性物质主要位于腺癌细胞质内,细胞核阴性。AMH在低分化腺癌的阳性表达率(30.0%)低于高分化腺癌(87.5%)和中分化腺癌(76.9%),差异均有统计学意义(P0.05);AMHR-Ⅱ在低分化腺癌的阳性表达率(20.0%)低于高分化腺癌(81.3%)和中分化腺癌(69.2%),差异均有统计学意义(P0.05)。结论:AMH及AMHR-Ⅱ与子宫内膜样腺癌的发生有一定关系,而且随着分化程度的降低,二者的表达呈递减趋势。     

基质金属蛋白酶-2及孕激素受体在子宫内膜癌组织中的表达及其意义

目的检测基质金属蛋白酶-2（MMP-2）和孕激素受体（PR）在子宫内膜癌组织中的表达，分析其在子宫内膜癌中表达的相关性以及与临床病理特征的关系，探讨MMP-2在子宫内膜癌发生、发展中的作用。方法采用免疫组化SP法检测45例子宫内膜癌和12例增生子宫内膜中MMP-2、PR的表达，以20例正常子宫内膜作为对照。结果MMP-2蛋白在正常子宫内膜、增生子宫内膜中的阳性表达率比较，差异无显著性（P〉0．05）。子宫内膜癌组织中MMP-2阳性表达率（62．2％）高于对照组（15．6％）（P〈0．05）。MMP-2在中、低分化子宫内膜癌组织中的阳性表达率（82．6％）高于高分化子宫内膜癌组织（40．9％）（P〈0．01），有肌层浸润标本中的阳性表达率（71．1％）高于无肌层浸润者（14．3％）（P〈0．05）；绝经者（65．5％）与未绝经者（56．3％）、手术病理为Ⅰ期者（52．0％）与Ⅱ～Ⅳ者（75．0％）以及有淋巴结转移者（63．6％）与无淋巴结转移者（61．8％）相比，MMP-2的阳性表达率差异无显著性。子宫内膜癌中PR的阳性表达率（55．6％）明显低于对照组（84．4％）（P〈0．05）。MMP-2与PR在子宫内膜癌中的表达呈负相关（P=0．0048，r=-0．42）。结论MMP-2在子宫内膜癌中呈高表达，可能在其进展中有重要作用，PR表达降低可能是MMP-2表达升高的原因；检测MMP-2在子宫内膜癌中的表达，有助于子宫内膜癌预后的判断。     

子宫内膜癌中雌激素受体亚型α第五外显子变异体的表达

目的:探讨雌激素受体亚型α的第5外显子野生型及变异体在内膜癌中的表达以及在子宫内膜癌发生发展中的作用。方法:选取1999年10月至2003年1月行手术治疗的子宫内膜癌患者41例,以2001年11月至2003年3月接受手术的正常子宫内膜癌患者29例做为对照,进行逆转录-多聚酶链反应(RT-PCR),半定量研究雌激素受体亚型第5外显子变异体的表达。结果:内膜癌组织中雌激素受体α第5外显子变异体的表达高于正常内膜组织(P<0.05),而二者之间野生型及野生型与变异体的比值差异无显著性(P>0.05)。结论:雌激素受体α第5外显子缺失型变异体可能在内膜癌的发生和发展中起重要作用。     

孕激素受体A、B亚型在子宫肌瘤中的表达

目的 旨在研究两种孕激素受体亚型 (hPR-A和hPR-B) 在子宫肌瘤和正常子宫肌层的分布及其mRNA的比例,探讨其在子宫肌瘤发生、发展中的意义。方法 选取22例因子宫肌瘤行全子宫切除术的标本,每1例取肌瘤组织和正常肌层组织,分别制成石蜡切片和冰冻标本。前者经免疫组化定位研究孕激素受体亚型;后者用于提取RNA,通过逆转录-多聚酶链反应,半定量研究孕激素受体亚型的mRNA表达。结果 免疫组化显示子宫肌瘤和正常肌层细胞的细胞核中,可见hPR及hPR-B的阳性颗粒;hPR-A、hPR-B和hPR-(A+B)的mRNA在肌瘤的表达量均高于周围正常肌层。无论在子宫肌瘤还是在正常肌层组织中,hPR-A与hPR-B的mRNA表达量无明显差异。结论 hPR-A、hPR-B为核受体;A亚型和B亚型的mRNA在子宫肌瘤表达高于正常肌层;孕激素受体亚型可能与子宫肌瘤的发生有关。     

nm23-H1基因在子宫内膜癌中的表达及其与临床预后的相关性研究

目的 研究ｎｍ２３－Ｈ１基因在子宫内膜癌中的表达,探讨其与了宫内膜癌临床各参数之间的相关性,了解ｎｍ２３－Ｈ１基因对子宫内膜癌预后的影响。方法 采用免疫组化染色链霉素亲合素生物素标记法（ＬＳＡＢ）检测子宫内膜癌６０例。结果 ｎｍ２３－Ｈ１基因在６０例子宫内膜癌中的阳性表达率为５５．０％。     

Comparative immunohistochemical study of endometrioid and serous papillary carcinoma of endometrium.

OBJECTIVE: The aim of this study was to determine whether immunohistochemical analysis of molecular parameters can provide an alternative method for classification of endometrial cancer cases according to their aggressiveness. METHODS: Sixty-four cases of endometrial carcinoma were assigned to three groups: group I--28 cases of endometrioid well and moderately differentiated (G1-G2) carcinoma; group II--14 cases of endometrioid poorly differentiated (G3) carcinoma; group III--22 cases of serous papillary endometrial cancer. Immunohistochemistry was used to determine the existence of estrogen receptors (ER), progesterone receptors (PR), and the expression of bcl-2, p53, HER-2/neu and Ki-67. RESULTS: There was a significant difference in the immunohistochemical profile of the studied molecular parameters comparing the three study groups. The endometrioid G1-G2 cases (group I) were characterized by increased immunoreactivity for ER and PR (85.7% and 78.6%, respectively), increased immunoreactivity for bcl-2 (42.8%) and low expression of p53 (14.3%) and HER-2/neu (14.3%). In contrast to group I cases, the serous papillary endometrial cancer cases (group III) were characterized by immunonegativity for ER, PR and bcl-2 and high immunoreactivity for p53 (81.8%) and HER-2/neu (45.4%). The endometrioid G3 cases (group II) demonstrated an intermediate immunoprofile, characterized by immunonegativity for ER, PR and HER-2/neu, low immunoreactivity for bcl-2 (7.1%) and high expression of p53 (57.1%). The expression of Ki-67 did not differ significantly comparing the different cases of endometrial cancer. CONCLUSION: This study provides evidence that the immunohistochemical analysis of endometrial carcinoma differentiates between different grades and histological types, thus being useful in the distinction of high risk cases.     

Correlation of androgen receptors with histological differentiation in human endometrial carcinomas

Primary endometrial carcinomas from 35 non-treated patients were investigated by measurement of androgen receptors in the tumor cytosols. Receptor analysis was done with a labelled synthetic androgen, methyltrienolone 3H-R1881 and triamcinolone acetonide, and dextran-coated charcoal absorption. The concentrations of androgen receptors in the endometrial carcinomas were, in decreasing order: highly-differentiated tumors, 15.7 +/- 1.8 (fmol/mg protein, mean +/- SE) (number of cases, 21); moderately differentiated tumors, 4.6 +/- 1.6 (n = 7); poorly differentiated tumors, undetectable in 7 out of 8 cases. Highly-differentiated tumors contained a much greater concentration of receptors than the two less differentiated ones. One highly-differentiated endometrial carcinoma with pyometra virtually lacked the receptor. The moderately differentiated endometrial carcinomas contained very low levels of the receptors. The poorly differentiated tumors virtually lacked the receptors. Metastatic lymph nodes from primary endometrial carcinomas with moderate differentiation had a very low receptor level. From these results, it is concluded that human endometrial carcinomas, particularly with histologically high differentiation, contain a considerable amount of androgen receptor and that the receptor concentrations appear to correlate with the histologic grade of tumor differentiation.     

S100A14在子宫内膜癌中的表达及其意义

目的:探讨S100A14在子宫内膜样腺癌中的表达及其临床意义。方法:采用免疫组化法检测子宫内膜癌组织、不典型增生子宫内膜组织、单纯性增生子宫内膜组织及正常子宫内膜组织中S100A14的表达情况。结果:84例子宫内膜癌患者中,高、中分化与低分化癌组织中S100A14表达具有统计学差异(P=0.020,P=0.034);行淋巴结清扫的74例子宫内膜癌患者中,淋巴结阴性与淋巴结阳性癌组织中S100A14表达有统计学差异(P=0.044)。子宫内膜癌组织中S100A14的表达与正常内膜组织比较,差异有统计学意义(P=0.034),单纯性增生、不典型增生子宫内膜组织中S100A14的表达与正常内膜组织比较,无统计学差异。结论:S100A14的表达下调与子宫内膜样腺癌的恶性程度及肿瘤转移程度相关。     

Progesterone receptor isoform identification and subcellular localization in endometrial cancer

OBJECTIVE: These studies were undertaken to characterize the subcellular localization of the two major isoforms of progesterone receptors (PR), PRA and PRB, in endometrial cancer. METHODS: Immunohistochemistry, immunoprecipitation, and confocal microscopy were performed using Hec50co and KLE endometrial cancer cell models expressing PRA or PRB as a consequence of transduction. The location of PRB compared to PRA was determined, and antibodies were tested for specificity with respect to PR isoform recognition. Immunohistochemical analyses of PR expression and subcellular compartmentalization were also performed on 20 formalin-fixed endometrial cancer tumors. RESULTS: Morphological and biochemical evaluations demonstrated that PRA is localized to the nucleus, even in the absence of progesterone. In contrast, a large proportion of PRB is cytoplasmic in the absence of ligand, but is rapidly translocated to the nucleus in the presence of progesterone. The differential distribution of PRA and PRB proved to be a hallmark of malignant and nonmalignant epithelia in 20 samples of archival endometrial tissue from women with the pre-operative diagnosis of endometrial cancer. All endometrial cancer specimens demonstrated cytoplasmic PRB in 50% or more of the cells, and five of the seven tumors that were moderately to poorly differentiated demonstrated no PRB staining in the nuclei. Nuclear PRB was significantly associated with increasing tumor differentiation (P = 0.031). CONCLUSION: In the absence of ligand, PRA is nuclear and PRB is largely cytoplasmic. This suggests that PRA may exert ligand-independent nuclear effects, while PRB may have nongenomic cytoplasmic actions in endometrial cancer cells.     

Growth potential of endometrial cancers assessed by a Ki-67 Ag/DNA dual-color flow-cytometric assay

Ki-67 is a monoclonal antibody directed against a nuclear antigen present only in proliferating cells. To assess the growth potential of uterine endometrial cancer, the population of cells in proliferating cycle (%PC) was examined with Ki-67, using flow cytometry. The %PC of 27.18 +/- 12.00% in 22 endometrial cancers was significantly higher than the 14.5 +/- 5.94% found in 28 normal endometrial tissues. In premenopausal endometrial tissue, the %PC in the proliferatory phase was significantly higher than the %PC found in the secretory phase. In endometrial cancers, an increase of %PC was found in cases with deep myometrial invasion, and the %PC was elevated in groups containing histologically poorly differentiated types when compared to groups of well-differentiated and moderately differentiated types. Sorted cells reactive with Ki-67 antibody were large and had a high nuclear/cytoplasmic ratio. On the bases of these results, it was concluded that a Ki-67 Ag/DNA dual-color assay would be useful to examine the growth fraction in endometrial carcinoma and that an increased growth fraction was related to deep myometrial invasion or poorly differentiated types.