应用末端氧核苷酰转移酶法研究化疗药诱导人肿瘤细胞的凋亡

目的 应用末端脱氧核苷酰转移酶尘（ＴｄＴ法）研究抗药物诱导人肿瘤细胞的凋亡,方法 应用ＴｄＴ法、ＤＮＡ凝胶电泳以及电镜、光镜等技术研究有关细胞的凋亡。结果 本研究表明,顺铂可以诱地人白血病的ＨＬ－６０细胞和经ＳＶ４０－Ｔ转染的人支气管上皮细胞的雕恨,并有很好的剂量和时间反应,同时还观察到,这些细胞的ｂｃｌ－２ｍＲＮＡ表达降低先于细胞凋亡的发生。应用ＴｄＴ法检测,在卡铂治疗的卵巢癌患者的肿瘤细胞中到     

应用末端脱氧核苷酰转移酶法研究化疗药诱导人肿瘤细胞的凋亡

应用末端脱氧核苷酰转移酶法（ＴｄＴ法）研究抗肿瘤药物诱导人肿瘤细胞的凋亡。方法应用ＴｄＴ法、ＤＮＡ凝胶电泳以及电镜、光镜等技术研究有关细胞的凋亡。结果本研究表明，顺铂可以诱导人白血病ＨＬ－６０细胞和经ＳＶ４０－Ｔ转染的人支气管上皮细胞的凋亡，并有很好的剂量和时间反应，同时还观察到，这些细胞的ｂｃｌ－２ｍＲＮＡ表达降低先于细胞凋亡的发生。应用ＴｄＴ法检测，在卡铂治疗的卵巢癌患者的肿瘤组织中观察到了凋亡细胞。结论ＴｄＴ法有可能用于检测经化疗诱导的人肿瘤细胞的凋亡，以及评价治疗过程中肿瘤细胞对治疗的反应。     

他莫昔芬和甲孕酮对人卵巢癌细胞系增殖和凋亡的影响

目的：研究他莫昔芬和甲孕酮对人卵巢癌细胞增殖和凋亡的影响。方法：用不同剂量的他莫昔芬和甲孕酮与人卵巢癌细胞系ＨＯ－８９１０体外培养９６ｈ，用苔盼蓝活细胞拒染法计活细胞数，免疫组化ＳＡＢＣ法检测癌细胞增殖核抗原（ＰＣＮＡ）表达情况，ＤＮＡ缺口原位末端标记方法（ＴＵＮＥＬ）检测细胞凋亡情况。结果：他莫昔芬和甲孕酮在０．１、１、１０ｕｍｏｌ均可使ＨＯ－８９１０活细胞数显著减少（Ｐ〈０．０１）；通过诱导细     

新辅助化疗对Ⅲ期非小细胞肺癌细胞凋亡和增殖的影响

目的：探讨新辅助化疗对非小细胞肺癌肿瘤细胞凋亡及对肿瘤细胞增殖的影响。方法：选取Ⅲ期非小细胞肺癌患者56例行新辅助化疗后并手术．并选取同期50例直接手术患者作为对照组。两组患者的手术标本．分别采用末端转移酶介导的dUTP切口末端标记法（TUNEL）行细胞凋亡检测，采用免疫组化标记链菌亲和素生物素法（LSAB），检测细胞增殖抗原Ki－67的表达。结果：新辅助化疗组肿瘤细胞凋亡指数（AI）均数为9．34％，对照组肿瘤细胞凋亡指数均数为5．27％．两组比较有显著差异（P〈0，001）。新辅助化疗组肿瘤细胞增殖抗原Ki－67阳性表达率均数为35．68％．对照组Ki－67阳性表达率均数为59.35％，两组比较差异显著（P〈0．001）。新辅助化疗组及对照组中，肿瘤细胞凋亡指数AI与增殖指数Ki-67的阳性表达均成负相关。结论：新辅助化疗能诱导Ⅲ期非小细胞肺癌肿瘤细胞的凋亡．并能抑制其增殖。     

新辅助化疗对乳腺癌细胞凋亡和增殖的影响

背景与目的：在实验研究中,已证实多种化疗药物都可引起乳腺癌细胞的凋亡。但在人体内进行有关化疗药物引起乳腺癌细胞凋亡的研究,还少有报道。本文研究新辅助化疗能否引起乳腺癌肿瘤细胞的凋亡以及对乳腺癌肿瘤细胞增殖的影响。方法：应用末端转移酶介导的dUTP切口末端标记法（TdT-mediated dUTP nick end labelling,TUNEL)及免疫组化的标记链菌亲和素生物素法（Labelled Streptavidin Biotin,LSAB),分别检测100例乳腺癌组织中肿瘤细胞的凋亡指数（Apoptotic index,AI)和乳腺癌肿瘤组织的增殖细胞核抗原（Proliferating cell nuclear antigen,PCNA)表达。结果：新辅助化疗组肿瘤细胞凋亡指数（AI）均数为7．47％,与对照组肿瘤细胞凋亡指数（AI）均数4．83％相比明显增高（P＜0．01）。新辅助化疗组肿瘤细胞增殖细胞核抗原（proliferation cell nuclear antigen,PCNA)阳性表达率均数为33．71％,与对照组PCNA阳性率均数51．52％相比明显降低（P＜0．01）。在新辅助化疗组及对照组两组病例中,肿瘤细胞凋亡指数（AI）与增殖细胞核抗原（PCNA）的阳性表达均呈负相关。结论：在人体乳腺癌组织中,新辅助化疗能诱导肿瘤细胞发生凋亡,并能抑制其增殖。     

华蟾素联合顺铂对卵巢癌细胞A2780增殖及凋亡的影响

[目的]研究华蟾素联合顺铂对卵巢癌细胞A2780增殖、凋亡的影响。[方法]采用四甲基偶氮唑盐微量酶反应比色法(MTT法)检测华蟾素联合顺铂对卵巢癌细胞A2780的半数抑制浓度(IC_(50))和细胞生长抑制率,采用流式细胞术检测细胞周期,Hoechst染色检测细胞凋亡。[结果]华蟾素及顺铂能抑制细胞增殖,华蟾素IC_(50)为0.56mg/ml;顺铂IC_(50)为2.8μg/ml;药物对癌细胞的生长抑制率随时间延长而增强。华蟾素和顺铂对A2780细胞表现出凋亡作用,华蟾素使A2780细胞周期停滞于S期(P<0.01),顺铂使A2780细胞周期停滞于G0/G1期(P<0.05)。[结论 ]华蟾素联合顺铂可诱导A2780细胞周期阻滞和细胞凋亡,从而抑制A2780细胞生长。     

宫颈癌中癌基因、细胞增殖和细胞凋亡的定位观察

宫颈癌的发展缓慢且多由癌前病变发展而来。本研究采用免疫组化技术及ＤＮＡ缺口末端标记(ＴＵＮＥＬ)技术对正常宫颈、宫颈上皮内瘤样病变(ＣＩＮ)和宫颈鳞癌患者的手术石蜡标本进行癌基因、细胞增殖和细胞凋亡状况的检测 ,同时对该组患者的临床及病理资料进行分析。1　材料与方法1.1　材料取自上海医科大学妇产科医院 1987年 1月至1994年 12月手术石蜡标本 4 7例 ,其中ＣＩＮⅠ～Ⅱ级 8例 ,ＣＩＮⅢ级 16例 ,宫颈鳞癌 18例 ,年龄 2 9～ 73岁 ,平均 56岁。同期同年龄段正常宫颈组织 5例作对照。1.2　方法检测ｐ53、ｃ ｍｙｃ表达和ＰＣＮＡ…     

流式细胞术检测化疗前后卵巢癌细胞凋亡的临床意义

目的：探讨化疗前后卵巢癌细胞及外周血淋巴细胞凋亡的因子Ｆａｓ、Ａｐｏ２．７的含量及其临床意义。方法：从７５例化疗前后卵巢癌患者的腹水中提取富含癌细胞的悬液,用Ａｎｎｅｘｉｎ Ｖ－ＦＴＴＣ／ＰＩ双染法分辨凋亡的亡细胞及坏死细胞及坏死细胞,并以两种凋亡因子克隆抗体Ｆａｓ和Ａｐｏ２．７标记上述细胞,均用流式细胞仪检测。结果：化疗后卵巢癌细胞凋亡率显著高于化疗前的临床疗效好,反之疗效差,被Ｆａｓ及Ａｐｏ２     

抗肿瘤药物与卵巢癌细胞凋亡

细胞调亡（Apoptosis,APO）存在于正常的胚胎发育过程中，是机体保持组织同一性的方式之一。近年来的研究表明，APO也存在于肿瘤的生物学领域内，并且已知几乎所有的抗肿瘤药物均能诱导敏感细胞的APO，且认为其本质就是诱导肿瘤细胞的调亡，而不是以前所公认的细胞坏死[1]。为此本文简要综述抗肿瘤药物与卵巢癌APO的关系及其基因调控。一、卯巢与卵巢癌的APO有作者发现不同发育阶段的正常卵泡并没有出现APO，但发育过程中印泡的闭锁是以APO的形式进行，并且其过程受促性腺激素和性腺激素的调控，雌激素促进卵泡的生长分化，阻止凋…     

格尔德霉素对人卵巢癌细胞SKOV3增殖和凋亡的影响及其机制探讨

目的:研究格尔德霉素(geldanamycin,GA)对人卵巢癌SKOV3细胞的体外抑制作用,探讨GA抑制SKOV3细胞增殖及诱导其凋亡的可能机制。方法:MTT法检测GA单独或联合顺铂(cisplatin)对SKOV3细胞的增殖抑制作用;FCM法检测GA作用后SKOV3细胞周期及凋亡率的变化;分别应用免疫细胞化学法(immunocytochemistory,ICC)和FCM法检测GA处理SKOV3细胞48h后细胞内Akt、Raf-1蛋白的表达变化。结果:GA对SKOV3细胞的体外增殖可产生明显的抑制作用(P<0.05),该作用随着GA浓度的增高和作用时间的延长而增强。而且,GA可增强顺铂对SKOV3细胞的增殖抑制作用,在一定浓度范围内随着GA浓度的增高,两药联合应用对SKOV3细胞的增殖抑制作用越强。经0～2000nmol/L GA作用SKOV3细胞48h后,G2/M期细胞逐渐增多,细胞凋亡率也显著升高(P<0.05)。ICC和FCM法检测结果均表明,经0～2000nmol/L GA处理48h后,SKOV3细胞中Akt、Raf-1蛋白的表达水平随着药物浓度增高而逐渐降低(P<0.05)。结论:GA能够对卵巢癌SKOV3细胞产生明显的体外增殖抑制作用,并诱导其凋亡,其作用机制可能与下调卵巢癌细胞中Akt和Raf-1蛋白表达相关。     

还氧合酶-2蛋白表达与卵巢癌生物学行为的相关性

目的　研究还氧合酶 - 2 (cyclooxygenase 2 ,COX 2 )蛋白表达与卵巢浆液性肿瘤发生发展的关系。方法　应用免疫印迹 (Westernblot)法对 5 4例卵巢浆液性肿瘤组织和 10例正常卵巢组织进行COX 2蛋白检测。结果　COX 2蛋白在卵巢浆液性癌组织中表达率 (81.8% )和相对含量(2 0 .0 8± 3.5 3)明显高于良性卵巢浆液性肿瘤 (0 ,15 .0 4± 0 .12 )及正常卵巢组织 (0 ,15 .33± 0 .6 0 ;P <0 .0 5 ) ;COX 2蛋白在交界性卵巢浆液性肿瘤中表达率 (90 .0 % )和相对含量 (2 0 .6 1± 3.0 3)明显高于良性卵巢浆液性肿瘤及正常卵巢组织 (P <0 .0 5 )。卵巢浆液性癌组织与交界性卵巢浆液性肿瘤、良性卵巢浆液性肿瘤组织与正常卵巢组织比较 ,COX 2蛋白表达率和相对含量差异均无显著性 (P >0 .0 5 )。COX 2蛋白表达率和相对含量在不同的临床分期 (Ⅰ +Ⅱ与Ⅲ +Ⅳ )、组织学分级、有无腹水及有无淋巴结转移间比较 ,差异均无显著性 (P >0 .0 5 )。结论　COX 2蛋白与卵巢浆液性肿瘤发生发展密切相关。COX 2可作为卵巢浆液性癌早期诊断指标及治疗靶位点     

凋亡与增殖及其调控基因表达在直肠癌术前放疗前后的对比研究

目的 :通过检测直肠癌术前放疗患者放疗前后凋亡、增殖指数以及凋亡调控基因表达的变化 ,探讨直肠癌自发性凋亡及放射性凋亡的可能调控机制。方法 :分析 2 3例接受术前放疗的直肠癌患者的临床资料。采用TUNEL法观察放疗前活检标本及放疗后手术切除标本的细胞凋亡情况。免疫组化方法检测所取标本的p5 3、p2 1、PCNA、bcl 2、bax等表达情况。结果 :放疗前活检标本中凋亡指数为 2 8± 1 4,而放疗后标本中凋亡指数为 4 9± 1 3 ,P <0 0 1。放疗后p5 3、p2 1、bcl 2、PCNA染色阳性率较放疗前均下降 ,而bax表达阳性率由放疗前的 3 4 8%上升到放疗后的 65 2 % ,但差异均无统计学意义 ,P >0 0 5。在放疗前活检标本中 ,p5 3 ( -)p2 1( )组的凋亡指数为 4 2± 1 0 ,高于p5 3 ( )p2 1( -)组的 2 3± 0 6,P <0 0 5。放疗后手术标本中 ,p5 3 ( -)p2 1( )组的凋亡指数为 5 8± 1 5 ,高于p5 3 ( )p2 1( -)组的 4 3±0 6,P <0 0 5。结论 :在直肠癌组织的自发凋亡和放射性凋亡过程中 ,p5 3 /p2 1途径均起到了关键作用 ,bax也可能起到一定的作用。     

宫颈癌中细胞增殖和细胞凋亡的关系研究

Objective To study the relationship between cell proliferation and apoptosis in cervical carcinoma and its clinical significance.Methods The cell proliferation and apoptosis of cervical epithelial cells in archival formalin-fixed,paraffin-embedded tissue sections of normal cervix ,cervical intraepithelial neoplasms(CN) and cervical squamous carcinoma were tested by using immunohistochemistry assay and DNA nick end-labeling technigue.The proliferation index(PI) and apoptosis index(AI) were calculated and their correlation with clinical and pathological data was analyzed. Results PI was gradually increased,but the AI and AI/PI ratio decreased from normal cervical epithelium,CIN to cervical carcinoma. There was no significant relationship among cell proliferation,apoptosis,clinical stages and pathological grades.High AI was always asso-ciated with a poor prognosis of the patients. Conclusion Cell proliferation and apoptosis allow to distinguish among normal epithelium,CIN and cervical carcinoma and are useful for the assessment of the malignant potential of tumor tissues.     

Combination chemotherapy with or without hexamethylmelamine in alkylating-agent resistant ovarian carcinoma

The study was designed to determine the efficacy of a two-drug and three-drug combination chemotherapy regimen for patients with advanced epithelial ovarian carcinoma resistant to alkylating monotherapy. Patients were randomized to receive either Adriamycin (doxorubicin) and cis-diamminedichloroplatinum(II) (AP) repeated every 3 weeks, or AP plus hexamethylmelamine (HAP) repeated every 5 weeks. Forty-five patients were evaluable for response and 49 for survival. No significant differences were found between the treatment groups as to response rate, progression-free survival, and survival. A remission was achieved in 20% of the patients and stable disease in another 20%. Median progression-free survival of all patients was only 4 months (median survival, 6 months). All patients showed progressive disease within 13 months after the onset of chemotherapy. Patients responding to treatment and those with an interval of more than 2 years between the initial diagnosis and cancer recurrence, experienced prolonged survival. Two conclusions can be drawn from the results of this study; neither of the regimens is superior to the other, and the effect of both in alkylator-resistant patients with ovarian cancer are meager. In studies on salvage chemotherapy, to the contrary, these combinations induced remissions in more than 40% of the patients. This difference in response rate might be due to differences between the prognostic factors of the patient populations. Better results are to be expected when these drugs are used in initial drug programs for previously untreated patients.     

High-dose chemotherapy in ovarian carcinoma

High-dose chemotherapy supported by bone marrow transplantation is being seriously investigated as a way to increase response and prolong survival in women with ovarian carcinoma. This report is a broad review of attempts over the past several years to demonstrate advantages of the high-dose approach. The existing data for dose escalation, short of doses requiring hematopoietic stem-cell transplant, demonstrate increased response rates, but no survival advantage. When even higher doses of chemotherapy are used in concert with autologous transplant to combat hematologic toxicities, even higher response rates have been reported. Despite this, all survival data from these later studies are limited. There is some hint of survival benefit in certain populations, such as those women with limited disease or perhaps in more advanced disease that has not been previously treated. All of the studies taken as a whole effectively demonstrate the need for randomized trials to address the issue of whether this approach provides any advantage over standard-dose chemotherapy. Future efforts at studying high-dose chemotherapy and autologous transplantation for ovarian carcinoma should be focused in this area; until results are available from such trials, high-dose chemotherapy with transplant will remain an experimental procedure.     

The effects of chemotherapy on morphology, cellular proliferation, apoptosis and oncoprotein expression in primary breast carcinoma.

The use of chemotherapy as a form of primary treatment for breast cancer is increasing and, as a result, more resection specimens contain tumours which have been exposed to cytotoxic drugs. We have studied the effects of chemotherapy on the tumour morphology and various biological features of breast carcinoma in a group of 35 patients. These were a group who responded to treatment in a clinical study of the use of primary chemotherapy designed to reduce tumour bulk prior to surgery. Characteristic morphological changes, temporally related to the administration of cytotoxic agents, are seen. The malignant cells become enlarged with vacuolated cytoplasm and vesicular nuclei containing prominent nuclei; occasionally the nuclei were angular and hyperchromatic. These features are interpreted as degenerative in nature. In 15 cases sufficient material was present in the pretreatment biopsies to compare the grade of the tumours before and after chemotherapy: changes were found in six tumours. Cytotoxic drugs do not induce a consistent pattern of change in the proliferation and apoptotic indices of individual tumours, but there is a tendency to reduce proliferative activity over all the tumours as a group. It was also found that chemotherapy is capable of modifying the expression of the oncoproteins c-erbB-2 and p53 in a minority of cases of breast cancer, usually resulting in an acquisition of immunoreactive oncoprotein. It is important to be aware of these effects when studying breast carcinomas removed after chemotherapy.     

钙拮抗剂增加化疗中卵巢癌细胞凋亡相关因子的研究

目的 探讨钙拮抗剂（ＣＡ）对促进化疗化物诱发卵巢癌细胞凋亡的作用。方法：从６８例首次手术及１７例复发卵巢癌患者腹水中提取富含癌细胞的悬液,应用３种荧光单克隆抗体Ｐ１７０,ＣＤ９５,及Ａｐｏ,２．７标记,经流式细胞仪检测,结果 应用ＣＡ组化疗后癌细胞ＣＤ９５及Ａｐｏ２．７含量均高于姐（Ｐ〈０．０５）,对癌细胞Ｐ１７０含量无明显影响（Ｐ〉０．０５）,复发组癌细胞Ｐ１７０含量显著高于初发手术组（Ｐ〈０．     

口腔鳞癌细胞增殖和细胞凋亡的相关性研究

目的：通过观察口腔鳞癌中细胞增殖和细胞凋亡之间的关系,探讨细胞凋亡在口腔鳞癌发生中的作用。方法：利用脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记（TUNEL）技术及增殖细胞核抗原（PC—NA）免疫组织化学染色,对69例（维族36例、汉族33例）口腔鳞癌中的凋亡细胞和增殖细胞进行原位观察和比较。结果：口腔鳞癌不同组织学分级比较,高分化口腔鳞癌与中低分化口腔鳞癌之间增殖指数及凋亡指数均有显著性差异（P〈0．05）,不同部位的口腔鳞癌其增殖指数无显著差异,而舌癌组凋亡指数低于唇癌组和牙龈癌组（P〈0．05）,增殖指数与凋亡指数在口腔鳞癌中呈负相关关系（r=-0．663,P〈0．05）。结论：口腔鳞癌癌变过程不仅存在活跃的细胞增殖,而且存在细胞凋亡之异常,细胞增殖和细胞凋亡平衡失调在舌癌发病中可能起重要作用。