子宫内膜增殖症与子宫内膜癌细胞凋亡的相关性研究

目的：探讨子宫内膜增殖症和子宫内膜癌细胞凋亡及其与凋亡相关基因产物表达的关系。方法：应用ＴＵＮＥＬ法检测正常增殖期、增殖症及癌变子宫内膜组织标本的凋亡细胞，应用组免法检测Ｂｃｌ ２、Ｐ５３、Ｆａｓ及Ｆａｓ Ｌ抗原。结果：（１）与正常增殖期相比，增殖症子宫内膜腺上皮细胞中凋亡细胞比率高，Ｂｃｌ ２ 蛋白含量高，Ｆａｓ、Ｆａｓ Ｌ蛋白含量低下；（２）内膜癌癌细胞中凋亡细胞比率高，Ｂｃｌ ２蛋白含量低，１／３标本中Ｆａｓ、Ｆａｓ Ｌ蛋白含量丰富；（３）Ｐ５３ 仅存在于１２％ 的内膜癌细胞核内。结论：增殖症内膜凋亡细胞增多显示了良性病变内膜中细胞凋亡对过度增生的抑制。内膜癌细胞凋亡增多则与Ｂｃｌ ２ 低表达，Ｆａｓ、Ｆａｓ Ｌ高表达及Ｐ５３部分表达相伴，表明癌变内膜中细胞凋亡调控受多种基因表达的影响。     

新生大鼠缺氧缺血性脑损伤时Bcl-2蛋白和Bax蛋白表达的研究

目的　研究Ｂｃ１－２蛋白和Ｂａｘ 蛋白在新生儿缺氧缺血性脑损伤（ＨＩＢＤ）中的表达,以及与细胞凋亡的关系。　方法　将新生７日龄Ｗｉｓｔａｒ大鼠制成ＨＩＢＤ模型,应用免疫组织化学——ＳＰ法及原位缺口末端标记（ＴＵＮＥＬ）。　结果　新生大鼠ＨＩＢＤ时细胞凋亡与坏死并存,但以凋亡为主。Ｂｃｌ－２和Ｂａｘ 在正常新生大鼠脑内广泛表达（＋ ～）;缺氧缺血（ＨＩ）后脑病变处Ｂｃｌ－２免疫强度明显下降（－ ～＋ ）;Ｂａｘ 的免疫强度也减弱,其中可见散在分布的阳性凋亡细胞;ＨＩ后Ｂｃｌ－２与Ｂａｘ 的比例下降。　结论　Ｂｃｌ－２可能抑制细胞凋亡,Ｂａｘ 可能诱发细胞凋亡     

细胞凋亡与Fas、Fas配体表达在子宫内膜癌中的意义

目的：研究子宫内膜癌的细胞凋亡Fａｓ、Fａｓ配体（ＦａｓＬ）表达其意义。方法：光镜下观察３５例子宫内膜癌及１５例子宫内膜增生ＨＥ染色石蜡切片中的凋亡细胞和凋亡小体，计算细胞凋亡指数（ＡＩ）。应用免疫组化ＳＡＢＣ法检测组织中Ｆａｓ及ＦａｓＬ的表达。结果：子宫内膜癌中细胞ＡＩ显著高于子宫内膜增生。在子宫内膜癌中高ＡＩ与高组织学分级、肌层浸润大于１／２及有淋巴结转移，患者生存率低有关。Ｆａｓ表达阳性率在子宫内膜癌中明显低于子宫内膜增生。ＦａｓＬ表达阳性率在子宫内膜癌中明显高于子宫内膜增生。结论：细胞凋亡异常在子宫内膜癌发病中起重要作用。ＡＩ对判断患者预后具有一定的价值。Ｆａｓ、ＦａｓＬ表达异常可导致子宫内膜癌发生、发展。     

子宫肌瘤与细胞凋亡关系初探

目的 探讨子宫平滑肌瘤的发生与细胞凋亡的关系。方法 采用免疫组化技术监测子宫肌瘤患者的肌瘤核组织（实验组）及其周围正常肌组织（对照组）中,ｐ５３基因蛋白产物、Ｆａｓ基因蛋白产物、Ｆａｓ配体（Ｆａｓ－Ｌ）基因蛋白产物及Ｂｃｌ－２基因蛋白产物的表达。结果 实验组各项阳性表达：ｐ５３蛋白为３６．６７％,Ｆａｓ－Ｌ蛋白为５６．６７％,Ｂｃｌ－２蛋白为７７．４２％;对照组各项阳性表达;ｐ５３蛋白为２０．００     

米非司酮影响早孕滋养细胞的信号传导和凋亡的机？…

目的 探讨米非司酮（ＲＵ４８６）在早孕滋养细胞信号传导及其凋亡的影响。方法 ２１９例服用ＲＵ４８６早孕药物流产者,对其有效组３０例,失败组２３例,人流组对照１６例,以免疫组化法分别标记孕激素受体（ＰＲ）、雌激素受体（ＥＲ）、表皮生长因子受体（ＥＧＦＲ）、酪氨酸蛋白激酶Ｃ（αＰＫＣ）、分裂激活蛋白激酶（ＭＥＫ－１）、细胞转化分裂介质（ｒａｆ－１）和抗凋亡蛋白（Ｂｃｌ－２）。三组均计数凋亡指数（ＡＩ）     

bcl-2、P~(53)、Fas蛋白表达与卵巢上皮癌细胞凋亡的相关性研究

目的：探讨ｂｃｌ－２、Ｐ５３、Ｆａｓ基因与卵巢上皮癌细胞凋亡的关系。方法：４４份卵巢癌术后石蜡包埋组织切片，应用原位末端标记法及免疫组化法进行相关检测。结果：卵巢上皮癌组织中有细胞凋亡和ｂｃｌ－２、Ｐ５３、Ｆａｓ蛋白的表达，阳性率分别为７５．００％、４３．１８％和４５．４５％。ｂｃｌ－２、Ｐ５３蛋白表达与卵巢上皮癌细胞凋亡呈负相关，Ｆａｓ蛋白表达与细胞凋亡呈正相关。上述４项指标在卵巢浆液性囊腺癌和卵巢粘液性囊腺癌之间差异无显著性（Ｐ＞０．０５）。结论：ｂｃｌ－２、Ｐ５３、Ｆａｓ基因可能通过调控细胞凋亡参与卵巢上皮癌的发生与发展。     

黄体细胞的凋亡

黄体退化时黄体细胞凋亡，其发生机制受ｃ－ｍｙｃ，Ｆａｓ及ＳＧＰ－２等基因表达的调控，也与ＴＮＦ－α，ＢＦＧＦ及ＩＦＮ－γ等细胞因子的作用有关。ｃ－ｍｙｃ，Ｆａｓ，ＴＮＦ－α及ＩＦＮ－γ促进细胞凋亡。ＢＦＧＦ等生长因子抑制细胞凋亡。     

异位子宫内膜细胞的凋亡与增殖的研究

目的 探讨异位子宫内膜细胞的凋亡特性及雌二醇（Ｅ２）、孕酮（Ｐ）、米非司酮对体外培养的人异位子宫内膜细胞表皮生长因子受体（ＥＧＦＲ）基因表达的影响。方法 对１５例子宫内膜异位症（内异症）患者的异位内膜组织（异位内膜组）及１１例非子宫内膜异位患者的在位内膜组织（在位内膜组）进行体外细胞培养,采用流式细胞术检测两组细胞的细胞凋亡指数,同时应用原位杂交方法检测细胞中ｂｃｌ－２的表达水平。应用逆转录聚合酶     

子宫内膜癌bcl-2癌基因的持续性表达及其临床意义

目的：研究子宫内膜癌ｂｃｌ－２癌基因的表达及其临床意义。方法：采用免疫组化ＡＢＣ法检测增生期、分泌期、单纯型增生、复合型增生及不典型增生子宫内膜共２６份，子宫内膜癌４９例的ｂｃｌ－２癌基因蛋白表达及雌、孕激素受体（ＥＲ、ＰＲ）的表达。结果：正常增生期子宫内膜、增生的子宫内膜存在ｂｃｌ－２的表达，与ＥＲ相关，分泌期子宫内膜ｂｃｌ－２表达下降；４９例子宫内膜癌中２６例ｂｃｌ－２表达阳性，占５３％，２９例ＥＲ表达阳性，占５９％，２５例ＰＲ表达阳性，占５１％。７２％ｂｃｌ－２表达阳性者ＥＲ阳性，７５％ｂｃｌ－２表达阴性者ＥＲ阴性（Ｐ＜０．０１）。６８％ｂｃｌ－２表达阳性者ＰＲ阳性，６２％ｂｃｌ－２阴性者ＰＲ阴性（Ｐ＜０．０５）。子宫内膜癌Ｇ１、Ｇ２级ｂｃｌ－２的表达率为６６％，显著高于Ｇ３级者（２１％）（Ｐ＜０．０５）。ｂｃｌ－２的表达与肌层浸润、手术分期无关，ｂｃｌ－２表达阳性及阴性者生存率统计差异无显著性。结论：子宫内膜ｂｃｌ－２的持续性表达与卵巢激素相互作用可能在子宫内膜癌发生、发展中起作用     

p53、bcl-2和bax基因表达与IUGR胎盘细胞凋亡的相关性研究

目的：研究胎儿宫内发育迟缓（ｉｎｔｒａｕｔｅｒｉｎｅ　ｇｒｏｗｔｈ　ｒｅｔａｒｄａｔｉｏｎ,ＩＵＧＲ)患者的胎盘细胞凋亡,以ｐ５３、ｂｃｌ－２和ｂａｘ相关性。方法：收集ＩＵＧＲ ２０份和正常足月分娩１２例的胎盘组织,应用原位末端标记(TUNEL)法检测其胎盘组织中的细胞凋亡,免疫组化法（ＩＨＣＡ）检测胎盘组织中ｐ５３、ｂｃｌ－２和ｂａｘ基因表达。结果：①正常和ＩＵＧＲ胎盘组织中均可见凋亡细胞,但ＩＵＧＲ胎盘中细胞凋亡指数（ａｐｏｐｔｏｓｉｓ　ｉｎｄｅｘ,ＡＩ）为１６．７～６８．３１,明显高于正常胎盘中的９．２～３０．４／高倍视野（平均２０．６７／高倍视野）,差异有显著性（Ｐ＜０．０５）;②ＩＵＧＲ ２０例胎盘组织中均有ｂａｘ过表达（４０％～９０％),高于正常组织（１０％～４０％）,差异有显著性（Ｐ＜０．０５）;③正常及ＩＵＧＲ胎盘组织中均未见ｂｃｌ－２表达;④ｐ５３在ＩＵＧＲ胎盘组织中的表达为５０％～８０％,而在正常组织中为５５％～８０％（Ｐ＞０．０５）;⑤ＩＵＧＲ２０例胎盘组织中细胞ＡＩ与ｂａｘ的表达呈正相关,与ｐ５３表达无关,ｂａｘ与ｐ５３之间无相关性。结论：①正常孕妇和ＩＵＧＲ患者胎盘组织中均有细胞凋亡,但ＩＵＧ     

孕激素及间质细胞培养液对原代培养内膜腺上皮HOXA11基因表达的影响

目的:探讨间质细胞是否参与了孕激素对子宫内膜腺上皮的调控,及其初步的作用机制。方法:将增生期子宫内膜间质细胞经激素处理后进行培养,提取培养液。用浓度为30%的提取培养液对腺上皮细胞进行原代培养,当细胞生长融合时,加入孕酮或孕雌激素培养4h、24h。提取细胞总RNA,用半定量RT-PCR方法检测腺上皮细胞HOXA11基因表达量。结果:当内膜腺上皮细胞中含有30%经孕激素处理的间质细胞培养液时,加入孕激素或孕、雌激素后其HOXA11基因,在培养4h时表达量有下降趋势;24h时,表达量下降明显;而用RU486预处理后再加入孕激素或雌孕激素,腺上皮细胞HOXA11基因表达量与对照组无差异;当上皮细胞中含有30%经RU486预处理后,再加入孕激素处理的间质细胞培养液时,孕激素或孕、雌激素对内膜腺上皮HOXA11表达的负调控作用在4h时消失;24h时,转为正调控(HOXA11基因表达量增加)。结论:孕激素对内膜腺上皮HOXA11基因的负调控作用需要问质细胞分泌的孕激素依赖因子的参与,而且由间质细胞和内膜腺上皮中的孕激素受体共同介导完成这一负调控作用。     

米非司酮及R_（2323）对离体培养人子宫内膜细胞PA与PAI－1生成的影响

孕酮拮抗剂米非司酮（ＲＵ４８６）在离体下对人子宫内膜间质细胞分泌组织型纤溶酶元激活因子（ｔＰＡ）及腺体细胞分泌尿激酶型纤溶酶元激活因子（ｕＰＡ）均有刺激作用，并抑制两种细胞分泌纤溶酶元激活因子的抑制因子（ＰＡＩ－１），其作用与孕酮相反。内美通（Ｒ２３２３）抑制ｔＰＡ和ｕＰＡ而刺激ＰＡＩ－１在二种细胞中的分泌，其作用与孕酮一致。提示ＲＵ４８６催经止孕、Ｒ２３２３抑制异位子宫内膜作用有可能是通过影响两种ＰＡ和ＰＡＩ—１的平衡介导的。     

米非司酮及R_（2323）对离体培养人子宫内膜细胞PA与PAI－1生成的影响

孕酮拮抗剂米非司酮（ＲＵ４８６）在离体下对人子宫内膜间质细胞分泌组织型纤溶酶元激活因子（ｔＰＡ）及腺体细胞分泌尿激酶型纤溶酶元激活因子（ｕＰＡ）均有刺激作用，并抑制两种细胞分泌纤溶酶元激活因子的抑制因子（ＰＡＩ－１），其作用与孕酮相反。内美通（Ｒ２３２３）抑制ｔＰＡ和ｕＰＡ而刺激ＰＡＩ－１在二种细胞中的分泌，其作用与孕酮一致。提示ＲＵ４８６催经止孕、Ｒ２３２３抑制异位子宫内膜作用有可能是通过影响两种ＰＡ和ＰＡＩ—１的平衡介导的。     

Mechanisms involved in the evolution of progestin resistance in human endometrial hyperplasia--precursor of endometrial cancer

BACKGROUND: Successful treatment of endometrial hyperplasia with progestins is commonly accompanied by the finding of an inactive or suppressed endometrium after therapy. However, approximately 30% of the endometrial hyperplasia cases do not respond to progestins and hyperplastic glands persist. The Fas/FasL system is known to play a role in tissue remodeling as a result of changes in menstrual hormone levels. The aims of this study are to examine Fas/FasL expression in endometrial hyperplasia of pre- and postprogestin treatment samples and to study the Fas/FasL regulation in vitro with Ishikawa cells after progestin stimulation. DESIGN: Pre- and posttreatment paraffin-embedded endometrial hyperplasia tissue samples from 26 women were examined by immunohistochemistry for changes in Fas/FasL expression related to the administration of progestins. Among 26 patients, 18 were successfully treated with progestins and 8 failed treatment. Fas/ FasL positivity was defined by the presence of 10% or more immunoreactive epithelial cells in each specimen. In positive cases, a percentage or an immunoscore of immunoreactive cells was given by counting 500 cells. Cell viability was evaluated by the MTT assay. The in vitro effects of progesterone on Fas/FasL expression and apoptosis in Ishikawa cells were examined by using Western blot and TUNEL assays, respectively. RESULTS: Fas immunoreactivity was present in 4/26 (15%) preprogestin cases with an average of 16% of the epithelial cells expressing Fas. FasL was expressed in 21/26 (80%) pretreatment cases with an average of 42% of the hyperplastic glandular cells being positive. In postprogestin cases, an increase of Fas expression (14/18, 77%) with an average of 47% stained cells was seen in responders (P < 0.001), while FasL was found in 16/18 (89%) responders with an average of 65% of cells positive (P = 0.587). In nonresponders, no significant changes in Fas/FasL expression were detected compared to pretreatment samples. With in vitro Ishikawa cells, a slight increase (10-20%) of Fas and FasL protein expression was detected after 24 h of progesterone treatment, but a more significant increase (220-343%) of both Fas and FasL expression was found after 48 h of withdrawing progesterone, which parallels apoptotic activity. CONCLUSIONS: The Fas/FasL system may be involved in the development of endometrial hyperplasia. Part of the molecular mechanisms of progestin therapy for endometrial hyperplasia is through upregulation of Fas/FasL expression. Dysregulation of Fas/FasL expression in hyperplastic endometrium may be part of the molecular mechanisms for nonresponders to progestin treatment. Intermittent, rather than continuous, progestin treatment may be more effective clinically for the treatment of endometrial hyperplasia.     

左旋18-甲基炔诺酮和米非司酮对分泌中期人子宫内膜HOXA11基因表达的影响

目的:通过口服孕激素类(左旋18-甲基炔诺酮,Levenorgestrel,LNG)和抗孕激素类(米非司酮,RU486)避孕药,研究孕激素对分泌中期人子宫内膜HOXA11基因表达的影响。方法:30名自愿者,于正常月经周期排卵后d7,刮取内膜组织做自身对照;实验周期在排卵前或排卵后2d,口服小剂量LNG(18例)或RU486(12例),同样在排卵后d7刮取内膜组织。用原位杂交方法检测服药子宫内膜HOXA11基因表达的变化。结果:对照组(分泌中期)内膜,间质细胞普遍表达HOXA11mRNA,而腺上皮HOXA11的表达则呈弱阳性或阴性。口服LNG后,子宫内膜形态变化不明显;内膜腺上皮HOXA11表达量进一步下降或无明显变化(即用药前后均表达阴性);间质细胞HOXA11表达增强。服用RU486后,内膜发育明显延迟,腺上皮HOXA11表达强度普遍大于对照组,而间质细胞的表达变化无明显规律。结论:孕激素对内膜腺上皮HOXA11基因的表达起负调控作用;正常分泌中期,内膜腺上皮HOXA11表达减弱或消失是内膜分化成熟的标志。     

正常人子宫内膜Fas、FasL协同表达与细胞凋亡相关性的研究

目的 :探讨正常人月经周期子宫内膜细胞凋亡及其与Fas基因家族的关系。方法 :应用TUNEL法检测细胞凋亡的时空分布 ,用原位杂交和免疫组化法检测Fas、FasL的表达。结果 :( 1)月经增殖晚期内膜无明显凋亡发生 ,Fas、FasLmRNA及蛋白的表达量极低 ;( 2 )月经分泌晚期内膜凋亡细胞比率高 ,Fas、FasLmRNA及蛋白的表达显著提高。结论 :Fas、FasL在人子宫内膜协同表达 ,且与子宫内膜凋亡的周期性变化一致 ,子宫内膜凋亡与Fas、FasL的高表达相关。内膜脱落、月经来潮可能是通过细胞凋亡实现的。     

离体培养下人子宫内膜细胞内皮素-1免疫活性物质的分泌与调节

本文应用免疫组化法观察人子宫内膜组织中的内皮素-1免疫活性物质(ET-1-ir)主要分布在腺细胞及腺腔上皮中,间质中分布很少.放免法(RIA)测得离体培养的人子宫内膜腺细胞培液中含有大量ET-1-ir,含量随培养时间延长而减少,并受雌、孕激素及RU486、dbcAMP、FK抑制,受佛波酯显著刺激而变化.间质细胞离体培养下几乎不分泌ET-1-ir.从而提示月经来潮、内膜脱卸及修复、增生可能与内膜腺体自分泌大量ET-1-ir有关.     

RU486-induced growth inhibition of human endometrial cells

OBJECTIVE: To determine the direct action of RU486 on endometrial cell proliferation and to differentiate whether the antioxidant or the antiprogesterone property of RU486 is predominately responsible for its effect on cell growth. DESIGN: In vitro study comparing the effects of RU486 (antiprogesterone and antioxidant), reduced RU486 (antioxidant), ZK112,993 (antiprogesterone), and lazaroid U74,500A (antioxidant) on endometrial cell growth. The human endometrial cell line EM42 was used in transient transfection assays to confirm the relative antiprogesterone potency of the various compounds. SETTING: Academic medical center PATIENT(S): Women presenting with pelvic pain or infertility and diagnosed with endometriosis at time of surgery or women desiring tubal ligation with a normal pelvis (controls). INTERVENTION(S): Endometrial cell cultures were treated with RU486, reduced RU486, lazaroid U74,500A, and ZK112,993. MAIN OUTCOME MEASURE(S): Tritiated thymidine incorporation was used to assess cell growth. Inhibition of progesterone induction of transiently transfected reporter plasmids was used to measure antiprogesterone activity of compounds studied. RESULT(S): RU486 reduced cell growth in a dose-dependent fashion of the endometrial cell lines EM42 and RL95-2 and of endometrial and endometriosis cells from primary culture. After being reduced, RU486 lost most of its antiprogesterone activity but retained its antiproliferative properties. ZK112,993 was similar in potency to RU486 as a progesterone antagonist but did not significantly modify endometrial cell growth. Lazaroid U74,500A was devoid of antiprogesterone activity but was shown to be a potent antiproliferative agent. CONCLUSION(S): RU486 has a direct inhibitory effect on human endometrial cell growth. This activity appears to be at least partly mediated through its antioxidant property.     

Inhibition of proliferation of endometrial stromal cells by trichostatin A, RU486, CDB-2914, N-acetylcysteine, and ICI 182780

BACKGROUND: All current major medications in treating endometriosis are effective in treating pain, most likely through suppression of proliferation of the implants, yet their effectiveness is relatively short term and they all have many undesirable, and sometimes severe, side effects. There is pressing need for novel, more effective medications in treating endometriosis with less and/or milder side effects. METHODS: Using a recently established immortalized endometrial stromal cell line, we carried out cell proliferation assays for cells treated with trichostatin A (TSA), RU486, CDB-2914, and N-acetylcysteine, and ICI 182780. Gene expression levels for PR-A, PR-B, AR, Fas and FasL were measured. Protein expression levels for ERalpha, ERbeta, and AR were also measured. RESULTS: Cell proliferation assay results for NAC, H2O2, CDB, and RU486 were nearly identical or similar to what have been reported based on primary cell cultures or in vivo studies. TSA, CDB, RU486 and NAC all had various antiproliferative effects. TSA had a more potent and longer lasting antiproliferative effect than CDB and NAC, even in the presence of an oxidant, H2O2. Its antiproliferative effect was concentration-dependent. ICI did not have a significant antiproliferative effect. PR-A, PR-B, AR, and FasL expression were all increased as compared with untreated cells. CONCLUSIONS: The cell line appears to be an adequate model for stromal components of endometriotic implants. That ICI has no inhibitory effect on endometrial proliferation may explain why a phase II clinical trial on its use to treat endometriosis did not advance to later stages. The upregulation of PR-B and AR may be responsible for antiproliferative effects induced by TSA, a histone deacetylase inhibitor (HDACI). HDACIs may be promising therapeutics in treating endometriosis due to their antiproliferative effects as well as the potential to restore gene dysregulation through chromatin remodeling.