癌基因bcl-2及抑癌基因p53在子宫内膜癌的表达及临床意义

目的：研究癌基因ｂｃｌ－２及抑癌基因ｐ５３在子宫内膜癌发生、发展中的作用。方法：采用免疫组化ＡＢＣ法检测４９例子宫内膜癌中ｂｃｌ－２、ｐ５３基因蛋白的表达。结果：４９例子宫内膜癌中２６例ｂｃｌ－２表达阳性，占５３％；１２例ｐ５３表达阳性，占２５％。子宫内膜癌组织学分级Ｇ１、Ｇ２ｂｃｌ－２表达率（６６％）显著高于Ｇ３表达率（２１％，Ｐ＜００５），而Ｇ３ｐ５３表达率（４６％）显著高于Ｇ１、Ｇ２表达率（１７％，Ｐ＜００５），ｂｃｌ－２表达阳性与阴性者生存率统计无显著性差异，ｐ５３表达阳性者生存率显著低于ｐ５３表达阴性者。ｂｃｌ－２、ｐ５３表达与肌层浸润、手术分期无明显相关。结论：癌基因ｂｃｌ－２与抑癌基因ｐ５３可能在子宫内膜癌发生的不同阶段起作用，抑制细胞凋亡，促进肿瘤的发展与转归。     

癌基因bcl—2及抑癌基因p53在子宫内膜癌的表达及临床 …

目的：研究癌基因ｂｃｌ－２及抑癌基因ｐ５３在子宫内膜癌发生，发展中的作用，方法：采用免疫组化ＡＢＣ法检测４９例子宫内膜癌中ｂｃｌ－２ｐ５３基因蛋白的表达，结果：４９例子宫内膜癌中的２６例ｂｃｌ－２表达阳性，占５３％１２例ｐ５３表达阳性，占２５％，子宫内膜组织学分级Ｇ１，Ｇ２，ｂｃｌ－２表达率（６６％）显著高于Ｇ３表达率（２１％，Ｐ〈０．０５），而Ｇ３ｐ５３表达率（４６％）显著高于Ｇ１，Ｇ２表达率（     

雌激素与三苯氧胺对子宫内膜癌细胞bcl-2 mRNA水平的调节

目的　探讨雌激素与三苯氧胺对子宫内膜癌ｂｃｌ２ 癌基因的调节机理。方法　采用逆转录－ 聚合酶链反应（ＲＴＰＣＲ） 法,检测子宫内膜癌细胞雌激素受体（ＥＲ） 阳性ＲＬ９５２ 细胞中ｂｃｌ２ｍＲＮＡ表达水平,及其与雌激素和三苯氧胺作用后的变化。结果　ＥＲ阳性ＲＬ９５２ 细胞有ｂｃｌ２ ｍＲＮＡ表达。经１０－１２ 、１０－１０ 、１０－８ 、１０－７ ｍｏｌ／Ｌ１７β雌二醇（Ｅ２）与ＲＬ９５２ 细胞作用７２ 小时后,ｂｃｌ２ 与βａｃｔｉｎｍＲＮＡ的比值分别为６．９４ ±０．０３、７ ．１５ ±０ ．０２、７．４７ ±０．０１、８．４４ ±０ ．０１ ,呈逐渐升高趋势;经１０－８ 、１０－７ 、１０－ ６、１０－５ ｍｏｌ／Ｌ三苯氧胺与ＲＬ９５２ 细胞作用７２ 小时后,ｂｃｌ２ 与βａｃｔｉｎ ｍＲＮＡ 的比值分别为３ ．６２±０．０３ 、２ ．８７ ±０．０１、２．２３±０ ．０１、１ ．９８ ±０ ．０２,呈逐渐下降趋势。结论　雌激素可通过促进ｂｃｌ２的表达,抑制细胞凋亡,在ＥＲ阳性的子宫内膜癌发病中起了一定作用;三苯氧胺则降低ｂｃｌ２ 的表达水平,促进细胞凋亡,可能产生治疗子宫内膜癌的效应。     

c－erbB2在增生性和恶性子宫内膜组织中的扩增与表达

目的：探讨子宫内膜癌的发生及发展与癌基因ｃ－ｅｒｂＢ２表达的关系。方法：应用差别聚合酶链反应（ＤＰＣＲ）和免疫组化法测定了２５例正常子宫内膜、３１例增生性子宫内膜和７２例子宫内膜癌组织中癌基因ｃ－ｅｒｂＢ２的扩增与表达。结果：正常子宫内膜仅有２例（２／２５，８．０％）低倍扩增，而子宫内膜增生及内膜癌组织中分别有１５和４５例（１５／３１，４８．４％；４５／７２，６２．５％）扩增，且扩增倍数从２～１２不等。免疫组化与ＤＰＣＲ法测定结果在阴性标本有很好的相关性。在子宫内膜增生中，不典型增生与复合增生的扩增率明显高于单纯性增生者。子宫内膜癌中，ｃ－ｅｒｂＢ２的高倍扩增率（≥５倍）与肿瘤的病理分级和癌细胞对血管或淋巴管侵入密切相关。结论：子宫内膜增生的癌变可能与ｃ－ｅｒｂＢ２的激活有关。高度激活的ｃ－ｅｒｂＢ２与内膜癌的发展、分化以及转移倾向密切相关，有可能作为临床预后的指标之一。     

在位内膜与异位内膜的分子生物学研究

近年来从分子生物学角度来分析在位内膜与异位内膜的差别的研究取得了显著进展，本文对细胞色素芳香化酶Ｐ４５０（Ｐ４５０ａｒｏｍ），血管内皮生长因子（ＶＥＧＦ），生长调节因子α（ＧＲα）及其受体，雌激素受体（ＥＲ）与孕激素受体（ＰＲ），ｂｃｌ－２，ｆａｓ以及Ｅｍ的免疫基础进行了综述。     

表皮生长因子受体在子宫内膜、早孕蜕膜及滋养细胞中的表达

目的了解表皮生长因子受体（ＥＧＦＲ）在子宫内膜细胞分化及发育中的作用。方法采用免疫组织化学及逆转录聚合酶链反应（ＲＴＰＣＲ）技术测定５８例正常子宫内膜（３２例增生期,２６例分泌期）与２６例早孕蜕膜。结果ＥＧＦＲ存在于正常子宫内膜、早孕蜕膜腺体及间质细胞的细胞膜、核膜及胞浆内,分布均匀。ＥＧＦＲ还位于早孕蜕膜腺体及间质细胞核内。正常子宫内膜ＥＧＦＲ的表达,腺体部分高于间质部分（Ｐ＜０．０５）,ＥＧＦＲ在增生期及分泌期子宫内膜腺体的表达差异无显著性（Ｐ＞０．０５）,但ＥＧＦＲ在早孕蜕膜组织中的表达明显高于增生期及分泌期（Ｐ＜０．０５）。ＥＧＦＲ在间质细胞的表达,早孕蜕膜高于分泌期内膜,而增生期内膜则表达最低（Ｐ＜０．０５）。ＥＧＦＲｍＲＮＡ的表达从弱到强依次为增生期、分泌期、蜕膜、滋养细胞,增生期与分泌期比较,差异无显著性（Ｐ＞０．０５）,早孕蜕膜较分泌期及增生期明显增加（Ｐ＜０．０５）,滋养细胞ＥＧＦＲｍＲＮＡ的表达明显高于增生期、分泌期及早孕蜕膜（Ｐ＜０．０５）。结论ＥＧＦＲ存在于各期子宫内膜中,其表达在正常月经周期中无明显变化,但在早孕蜕膜、滋养细胞中的表达明显高于增生期及分泌期内膜     

卵巢子宫内膜样癌HER－2／neu的表达

目的研究卵巢子宫内膜样癌ＨＥＲ－２／ｎｅｕ的表达以及与临床病理特征和预后的关系。方法应用ＨＥＲ－２／ｎｅｕ单克隆抗体免疫组化技术对卵巢子宫内膜样癌２８例进行ＨＥＲ－２／ｎｅｕ癌基因蛋白测定。结果１３例细胞浆染色阳性，占４６．４％，Ⅰ期肿瘤阳性率为２０．０％，Ⅱ～Ⅳ期肿瘤阳性率为５２．２％，手术有残留肿瘤者ＨＥＲ－２／ｎｅｕ的表达率是６２．５％，无残留肿瘤者ＨＥＲ－２／ｎｅｕ表达率是２５．０％，差异有显著性（Ｐ＝０．０４９）。ＨＥＲ－２／ｎｅｕ表达与发病年龄、ＣＡ１２５水平、组织学分级及淋巴转移无关。术后随诊６～９６个月，平均３１．３个月，ＨＥＲ－２／ｎｅｕ阳性者术后死亡率是３８．５％，ＨＥＲ－２／ｎｅｕ阴性者术后死亡率是２１．４％。结论ＨＥＲ－２／ｎｅｕ阳性者与卵巢子宫内膜样癌的不良预后有一定的关系。     

卵巢子宫内膜样癌HER－2／neu的表达

目的研究卵巢子宫内膜样癌ＨＥＲ－２／ｎｅｕ的表达以及与临床病理特征和预后的关系。方法应用ＨＥＲ－２／ｎｅｕ单克隆抗体免疫组化技术对卵巢子宫内膜样癌２８例进行ＨＥＲ－２／ｎｅｕ癌基因蛋白测定。结果１３例细胞浆染色阳性，占４６．４％，Ⅰ期肿瘤阳性率为２０．０％，Ⅱ～Ⅳ期肿瘤阳性率为５２．２％，手术有残留肿瘤者ＨＥＲ－２／ｎｅｕ的表达率是６２．５％，无残留肿瘤者ＨＥＲ－２／ｎｅｕ表达率是２５．０％，差异有显著性（Ｐ＝０．０４９）。ＨＥＲ－２／ｎｅｕ表达与发病年龄、ＣＡ１２５水平、组织学分级及淋巴转移无关。术后随诊６～９６个月，平均３１．３个月，ＨＥＲ－２／ｎｅｕ阳性者术后死亡率是３８．５％，ＨＥＲ－２／ｎｅｕ阴性者术后死亡率是２１．４％。结论ＨＥＲ－２／ｎｅｕ阳性者与卵巢子宫内膜样癌的不良预后有一定的关系。     

c—erbB2在增生性和恶性子宫内膜组织中的扩增与表达

目的：探讨子宫内膜癌的发生及发展与癌基因ｃ－ｅｒｂＢ２表达的关系。方法：应用差别聚合酶链反应（ＤＰＣＲ０和免疫组化法测定了２５例正常子宫内膜、３１例增生性子宫内膜和７２例子宫内膜癌组织中癌基因ｃ－ｅｒｂＢ２的扩增与表达。结果：正常子宫内膜仅２例（２／２５，８．０％）低倍扩增，而子宫内膜增生及内膜癌组织中分别有１５和４５例（１５／３１，４８．４％；４５／７２，６２．５％）扩增，且扩增倍数从２－１２不     

子宫内膜异位症患者内膜雌,孕激素受体测定及口服棉酚后的变化

采用葡聚糖活性碳吸附法，对２７例正常生育妇女（对照组）及２４例子宫内膜异位症患者（Ｅｍ组），测定了宫腔内膜雌激素胞浆受体（ＥＲｃ）、孕激素胞浆受体（ＰＲｃ）浓度。并测定Ｅｍ组中１５例口服醋酸棉酚２个月后，月经周期同日测定ＥＲｃ及ＰＲｃ浓度；同时对１６例手术切除的子宫、附件标本的原位内膜与异位内膜的ＥＲｃ、ＰＲｃ浓度进行比较。结果：对照组子宫内膜ＥＲｃ、ＰＲｃ浓度呈周期性变化，以卵泡晚期最高，黄体中期显著下降（Ｐ＜０．０５），黄体晚期进一步降低。Ｅｍ组原位内膜ＥＲｃ、ＰＲｃ浓度及周期性波动与对照组无差异，但异位内膜ＥＲｃ、ＰＲｃ浓度显著低于原位内膜（Ｐ各＝０．００３６及＜０．０００１），且失去周期性变化。口服棉酚２个月后血清雌二醇、孕酮浓度无显著下降，４例（２６．７％）患者子宫内膜组织相显示腺体分泌减少，但原位内膜ＥＲｃ、ＰＲｃ浓度却已显著降低（Ｐ＝０．０２１７及０．０１）。本研究提示：异位内膜与原位内膜对内源性性激素反应的不同步现象与异位内膜雌、孕激素受体浓度低下有关；棉酚对原位内膜ＥＲｃ、ＰＲｃ浓度的早期抑制现象，并不依赖于对卵巢功能的抑制作用，这一作用途径应对异位内膜同样有效。     

Persistent expression of bcl-2 onco-protein in endometrial carcinoma correlates with hormone receptor positivity

The protein product of proto-oncogene bcl-2 is thought to be involved in inhibition of apoptosis and is hormonally regulated in a variety of in vitro and in vivo experiments. The association of bcl-2 persistence and hormone receptor status was investigated by immunocytochemistry from paraffin-embedded tissue in a series of 82 women with endometrial carcinoma and 20 women with benign endometrium. In benign endometrium, bcl-2 immunoreactivity was strongly present in glands of proliferative and hyperplastic endometrium, while a weak signal was detected in secretory endometrium. Bcl-2 expression tends to decrease in staining intensity with progression from benign endometrium, including proliferative and hyperplastic endometrium, to endometrioid carcinoma and to mucinous, clear cell and serous carcinomas of the endometrium. Bcl-2 persistence was observed in the majority (65%) of endometrial carcinomas. We demonstrated a significant correlation of bcl-2 immunoreactivity with estrogen receptor ( P = 0.000005) and progesterone receptor status ( P = 0.00032). The bcl-2 persistence was found to be significantly higher in FIGO G1 and G2 tumors than in G3 tumors ( P = 0.00035), while no significant difference was detected in tumors of different stages. We conclude that bcl-2 persistence is highly correlated with the presence of hormone receptors and may be hormone-dependent or related to hormonal regulation in endometrial carcinomas. Persistent expression of bcl-2 in normal and hyperplastic endometrium and endometrial carcinoma suggests that failure to inactivate bcl-2 expression early in the development of endometrial carcinoma may provide an opportunity for accumulating genetic mutations and evolution from a precursor lesion to invasive carcinoma.     

Apoptosis, bcl-2 expression, and proliferation in benign and malignant endometrial epithelium: An approach using multiparameter flow cytometry

OBJECTIVE: Disturbances in the regulation of cell proliferation and differentiation play an important role in the formation of neoplastic lesions. Consequently, abnormalities in apoptosis regulation may contribute to this process. Expression of a neoepitope on cytokeratin 18, unmasked by an early caspase cleavage event and recognized by the novel monoclonal antibody M30, is an indicator of early epithelial cell apoptosis. The purpose of this study was to evaluate the quantitative relation among apoptosis (M30), cell persistence (bcl-2), and proliferation (S-phase fraction; SPF) in malignant and benign endometrium. METHODS: Using multiparameter DNA flow cytometry on 54 formalin-fixed paraffin-embedded samples from benign (proliferative, secretory, inactive, and hyperplastic endometrium) and malignant (grades 1-3 endometrial adenocarcinoma) endometrial tissue, bcl-2 expression and M30 reactivity were assessed together with the SPF in the cytokeratin-positive epithelial cells. RESULTS: Benign cyclic endometrium showed a relatively high bcl-2 expression and low M30 reactivity in the proliferative phase whereas in the secretory phase this relation was inverse. In endometrial hyperplasia the expression of bcl-2 was increased compared to that in secretory and postmenopausal endometrium, but still below the level of proliferative samples. The expression of M30 also increased compared to normal proliferative endometrium but did not reach the level of endometrium in the secretory phase of the menstrual cycle. In cancer the expression of bcl-2 decreased with the progression of differentiation grade. For M30 expression this relation was inverse. Overall there was a significant increase of M30 reactivity in cancerous compared to hyperplasia and normal cyclic endometrium. CONCLUSION: Transition of endometrial epithelium from hyperplasia to cancer seems to involve both increased apoptosis and decreased bcl-2 expression. Flow cytometric evaluation of M30 and bcl-2 expression levels, with the SPF, in currettage specimens from postmenopausal patients complaining of bleeding provides a quantitative assessment of endometrial apoptosis, anti-apoptosis, and proliferation. Further studies are needed to determine the relationship among these three processes as indicators of the biological behavior of gynecological tumors.     

PELP1在Ⅱ型子宫内膜癌组织中的表达及其与ER相关性研究

目的:探讨脯氨酸、谷氨酸和亮氨酸富集蛋白1(PELP1)在II型子宫内膜癌中的表达及其与雌激素受体(ER)的相关性。方法:免疫组化法检测84例子宫内膜癌及40例正常子宫内膜组织中PELP1和ER表达水平并对其进行相关性分析。结果:I型子宫内膜癌及增殖期子宫内膜组织中PELP1、ER阳性表达率高于II型子宫内膜癌及分泌期子宫内膜组织,差异均有统计学意义(P0.05);II型子宫内膜癌与分泌期子宫内膜组织比较,差异均无统计学差异(P0.05)。Pearson相关分析提示,PELP1与ER在I型子宫内膜癌组织中表达量呈正相关(rs=0.348,P=0.01),而在II型子宫内膜癌组织中无相关性(rs=0.268,P=0.144)。结论:PELP1在II型子宫内膜癌组织中的表达较少,且与ER表达无明显相关性。     

p16、Cyclin D1在增生性子宫内膜及子宫内膜癌中的表达

目的 :探讨 p16和 Cyclin D1在内膜癌发生、发展中的作用。方法 :采用免疫组化 S—P法对 12例正常子宫内膜、2 2例增生性子宫内膜及 4 1例子宫内膜癌中 p16和 Cyclin D1表达进行了研究。结果 :在单纯加复合增生、不典型增生及子宫内膜癌中 ,p16表达呈下降趋势 ,内膜癌与正常内膜及增生性内膜有显著性差异 (P<0 .0 1,P <0 .0 5 ) ;而Cyclin D1表达呈上升趋势 ,增生性子宫内膜、子宫内膜癌与正常内膜有显著性差异 (P<0 .0 5 ,P<0 .0 1)。不典型增生与单纯加复合增生 Cyclin D1过表达有显著性差异 (P<0 .0 1)。子宫内膜癌中 ,p16表达随细胞分化程度下降而降低 ,而Cyclin D1则随分化程度下降而上升 ,二者呈负相关。结论 :p16、Cyclin D1异常参与子宫内膜癌的发生 ;p16低表达、Cyclin D1过表达与内膜癌的恶性生物学行为有关 ;Cyclin D1核过表达可能是一个早期分子事件     

Heparanase expression increases throughout the endometrial hyperplasia-cancer sequence.

OBJECTIVE: To assess the expression of heparanase in the different stages leading to endometrial cancer. METHODS: The 38 examined specimens included adenocarcinoma, hyperplasia, and normal endometrium specimens. Heparanase, estrogen, and progesterone receptor expressions were analyzed immunohistochemically and the intensity was scored. RESULTS: Secretory normal endometrium and simple hyperplasia specimens expressed the lowest mean values of expression (1.00 and 0.63, respectively); the complex hyperplasia specimens and G2 endometrioid adenocarcinoma showed the highest values of expression (2.33 and 2.71, respectively). A linear trend (P=0.005) of heparanase expression was observed when comparing the normal endometrium and simple hyperplasia group with the complex hyperplasia+G1 carcinoma group and the G2+G3 carcinoma group. Evaluation of atrophic and inactive endometrium compared with papillary serous carcinomas yielded no significant differences. We found no significant correlation between heparanase expression and estrogen receptor or progesterone receptor expression. CONCLUSION: Heparanase expression was tightly regulated in endometrial tumorigenesis.     

细胞凋亡调控蛋白bcl-2和bax在多囊卵巢综合征患者子宫内膜中的表达

目的探讨多囊卵巢综合征（PCOS）患者子宫内膜中细胞凋亡调控蛋白bcl-2和bax的表达及其在PCOS患者子宫内膜病变中的意义。方法应用免疫组织化学SP方法,测定18例PCOS患者子宫内膜和27例月经周期正常、因输卵管不通或男性不育就诊的对照者子宫内膜（10例增殖期、17例分泌期）bcl-2和bax蛋白的表达。结果PCOS患者子宫内膜bcl-2表达较对照者增殖期和分泌期高（P＜0．05和P＜0．01）,但bax表达相似（p＞0．05）。结论PCOS患者子宫内膜bcl-2蛋白表达增高,与bax形成更多的异源二聚体,从而抑制PCOS子宫内膜凋亡,对其子宫内膜的异常增生改变起一定作用。     

Down-regulation of bcl-2 is a potential marker of the efficacy of progestin therapy in the treatment of endometrial hyperplasia

OBJECTIVE: The objective of this study was to evaluate the expression of bcl-2, a regulatory protein in programmed cell death, in endometrial hyperplasia before and after progestational therapy. METHODS: Pre- and posttreatment paraffin-embedded endometrial tissue samples from 20 women with an initial diagnosis of endometrial hyperplasia were obtained from archived files. Cases were evaluated and classified as either complete resolution of hyperplasia or persistent hyperplasia in response to progestin treatment. Sections were examined for bcl-2, estrogen receptor, and progesterone receptor expression using immunohistochemistry and compared within the treatment response groups. RESULTS: Among the 20 women studied, 13 had complete regression of their hyperplasia with progestin treatment and 7 had evidence of persistent disease after therapy. Bcl-2 expression was significantly decreased after treatment from a mean reactivity score of 2.08 to 0.31 (P = 0.0005) in the group of patients whose hyperplasia completely regressed with progestin administration. Among the women who had persistent hyperplasia after therapy, no significant change was observed between pre- and posttreatment bcl-2 expression, with a mean reactivity of 1.86 to 1. 29, respectively (P = 0.075). Progestational therapy significantly decreased the status of estrogen receptors from a mean score of 2.08 to 0.46 (P = 0.0005) in completely resolved cases of hyperplasia and from 2.00 to 0.43 (P = 0.0025) in persistent hyperplasias. Treatment also significantly decreased the status of progesterone receptors from a mean reactivity score of 1.92 to 0.31 (P = 0.0005) in cases of regressed hyperplasia and from a mean reactivity of 1.86 to 0.29 (P = 0.005) in persistent cases of hyperplasia. CONCLUSIONS: Bcl-2 expression decreases following successful progestin treatment of endometrial hyperplasias, whereas it remains expressed in hyperplasias which persist despite progestational therapy. This suggests that bcl-2 expression may represent a component of the therapeutic effects exerted in the endometium during progestational therapy in the treatment of hyperplasia. The activity of the oncoprotein may be a potential measure of the progress of treatment.     

子宫内膜增生性疾病患者内膜细胞凋亡的研究

目的 :研究凋亡在子宫内膜增生性疾病中的作用。方法 :用改良原位末端标记技术检测 15例正常月经周期的增生期、分泌期、月经期子宫内膜 ,11例增殖性子宫内膜 ,12例子宫内膜癌 ,以及术前用孕激素治疗的 13例异常增生子宫内膜中的凋亡细胞 ,并计算其凋亡指数 (AI)。结果 :分泌期、月经期子宫内膜、增殖性子宫内膜、子宫内膜癌AI均比正常增生期子宫内膜AI高 (P <0 .0 1)。增殖症患者内膜不典型增生组AI比单纯增生、复杂增生组AI高 (P <0 .0 5 ) ;内膜癌患者低分化组AI比高分化组、中分化组AI高 (P <0 .0 5 )。结论 :细胞凋亡与正常子宫内膜周期性变化有关 ,而在增殖性和癌变子宫内膜中的异常表达可能与子宫内膜的良恶性病变有关     

子宫内膜癌中p~(16)蛋白表达的免疫组化检测及其临床意义的研究

目的：探讨ｐ１６蛋白在子宫内膜癌中的表达及其临床意义。方法：应用抗ｐ１６抑癌基因的表达产物ｐ１６蛋白的单克隆抗体对５０份子宫内膜癌常规石蜡标本进行ｐ１６蛋白的免疫组化测定。结果：ｐ１６蛋白的表达广泛见于正常子宫内膜腺体、平滑肌和部分病例的癌组织中，阳性部位主要在细胞浆内。ｐ１６蛋白在子宫内膜癌表达的阳性率仅为４８．００％（２４／５０），其表达与子宫内膜癌的细胞分级、转移及患者预后有关（Ｐ＜０．０５），ｐ１６蛋白阴性者子宫内膜癌细胞分化程度低，易转移，患者预后差。结论：ｐ１６蛋白作为细胞周期的反向调节因子具有抑制细胞增殖的作用，ｐ１６抑癌基因的突变或缺失导致ｐ１６蛋白合成障碍，可引起细胞无限制性生长，即恶变。对子宫内膜癌中ｐ１６蛋白表达的测定有助于确定肿瘤的细胞分化程度，判断其侵蚀、转移的能力，并估计患者的预后。