多原发癌组织中bcl-2表达状况的研究

国内对多原发癌的研究相对较少,尚未见有对多原发癌分子病理学研究的报道[1～3].以往的研究表明,bcl-2基因的过表达在许多单发肿瘤的发生发展中起很重要作用[4～6],本实验应用免疫组织化学方法对多原发癌组织中的bcl-2蛋白表达进行检测.     

氟对软骨细胞凋亡因子bcl-2和Bax蛋白表达的影响

目的 观察氟对体外培养软骨细胞中凋亡相关因子bcl-2和Bax蛋白表达的影响,探讨氟在致软骨细胞凋亡中的作用机制.方法 采用软骨细胞体外培养方法,原代培养乳鼠关节软骨细胞,传第3代后按染氟剂量不同分为0(对照)、5、20、40mg/L组,培养10 d后,透射电镜下观察软骨细胞的超微结构改变,采用Western印迹法检测软骨细胞bcl-2和Bax蛋白表达.结果 透射电镜下,对照组和5 mg/L组软骨细胞呈球形,粗面内质网发达,线粒体膜性结构完整;20、40 mg/L组软骨细胞内可见脂滴增多,胞质内出现大量空泡类物质,细胞内膜结构不清,部分细胞出现核固缩.20、40 mg/L组软骨细胞bcl-2蛋白表达(0.626±0.042、0.531±0.039)较对照组(0.876±0.035)明显降低(P均＜0.01);而Bax蛋白表达(0.966±0.047、1.289±0.156)较对照组(0.642±0.050)明显增加(P均＜0.01).5 mg/L组软骨细胞bcl-2、Bax蛋白表达(0.885±0.065、0.657±0.045)与对照组比较,差异无统计学意义(P均＞0.05).此外,40 mg/L组与20 mg/L组比较,bcl-2、Bax蛋白表达差异有统计学意义(P均＜0.01).结论 染氟20、40 mg/L可对软骨细胞超微结构造成损伤,其通过减少抑凋亡因子bcl-2的表达和增加促凋亡因子Bax的表达,从而产生促进软骨细胞凋亡的作用.

Abstract：

Objective To study the effect of fluoride on the expression of bcl-2 and Bax in chondrocyte in vitro, and investigate the mechanism of action of chondrocyte apoptosis induced by fluoride. Methods Articular chondrocytes of neonate rat were cultured in vitro and treated with 0(control),5,20,40 mg/L of fluoride,respectively, for 10 days. Then observed the u]trastructure of chondrocytes under eletronicmicroscope, and tested the expression of bcl-2 and Bax in chondrocyte in different groups by Western blotting. Results Abundant rough endoplasmic reticulums (RERs) and complete structure of mitochondria membranes were presented in globular chondrocytes in the control group and 5 mg/L group; but more lipid droplets and vacuoles were seen in the cytoplasm, and the structure of intracellular membranes became incomplete, and some shrieked chromatin and pyknosis were seen in the chondrocytes of the 20,40 mg/L groups. The expression of bcl-2 markedly decreased in 20 mg/L group(0.626 ± 0.042) and 40 mg/L group(0.531± 0.039) compared to the control group(0.876 ± 0.035,all P ＜ 0.01 ). And the expression of Bax significantly increased in 20 mg/L group(0.966 ± 0.047) and 40 mg/Lgroup ( 1 .289 ± 0.156) compared to the control group(0.642 ± 0.050, all P ＜ 0.01). But there was no statistical significant difference of the expression of bcl-2 or Bax between 5 mg/L group(0.885 ± 0.065,0.657 ± 0.045) and control group (all P ＞ 0.05 ). However there were statistical differences of expressions of bcl-2 and Bax between 20 and 40 mg/L groups(all P ＜ 0.01 ). Conclusions Twenty and 40 mg/L fluoride can cause damage to the ultrastructure of chondrocyte, and fluoride possibly promotes chondrocyte apoptosis by reducing the expression of antiapoptotic factor bcl-2 and increasing the expression of Bax.     

离体肾脏保存过程中bcl-2蛋白、bcl-2 mRNA表达变化及FK506对其的影响

目的探讨离体肾脏保存过程中bcl-2蛋白、bcl-2mRNA表达及他可莫司（FKS06）对其的影响。方法将40只Wistar雄性大鼠随机分为实验组和对照组各20只，建立离体肾脏模型。对照组将离体肾脏置于常规4℃ UW保存液中；实验组在上液中加入FK506，分别检测肾脏保存2、4、8、16h后bcl-2蛋白、bcl-2m RNA表达。结果实验组4、8、16h后bcl-2蛋白、bcl-2mRNA表达明显高于对照组（P〈0．05）。结论FK506可能通过上调bcl-2蛋白、bcl-2mRNA表达，减轻离体肾脏保存过程中的损伤。     

直肠癌组织中bcl-2和PCNA表达的临床意义

1994年5月至1998年3月,我们应用免疫组化染色技术,检测68例直肠癌组织中bcl-2、增殖细胞核抗原(PCNA)的表达,并探讨其与预后的关系及临床意义。     

环氧化酶-2、bcl-2在68例胃癌中的表达及其与凋亡相关性

目的研究环氧化酶-2（cox-2）和bcl-2在68例胃癌中的表达及其与胃癌细胞凋亡的关系。方法采用SP免疫组织化学法检测胃癌及癌旁组织各68例标本中cox-2、bcl-2的表达，用脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记法（TUNEL）检测胃癌及癌旁组织的凋亡率。结果胃癌组织的cox-2、bcl-2的表达明显高于癌旁组织（P〈0．05），cox-2阳性组胃癌组织的凋亡率显著低干阴性组；cox-2的表达与bcl-2呈正相关。结论cox-2、bcl-2的异常表达与胃癌细胞凋亡抑制有关；cox-2、bcl-2能够使胃癌细胞凋亡抑制。     

转bcl-2治疗缺血性脑血管病的研究进展

Bcl 2抑制缺血性神经元凋亡的作用已引起人们的关注。近年来 ,利用转基因手段使bcl 2表达增加 ,能保护局灶性缺血性神经元免于凋亡 ,若方法适当 ,有可能从基因水平对缺血性脑血管病的防治提供良好的临床应用策略。     

急慢性肝病患者全血及血清中bcl-2的检测及其临床意义

目的:研究与细胞凋亡相关的bcl-2在急慢性肝病患者全血中存在情况及其临床意义,以及判断是否在血清中存在bcl-2;方法:选择46例患者为研究对象,其中男37例,女9例,年龄17岁～67岁,平均38.1岁。急性肝炎4例,慢性肝炎轻度8例,中度4例(慢性肝炎均经病理证实),肝炎肝硬变20例,慢性重型肝炎10例。乙型肝炎41例,丙型肝炎     

依那普利对肝组织bax和bcl-2基因表达的影响

目的探讨血管紧张素转化酶抑制剂（ACEI）依那普利（Ena）对大鼠肝纤维化的防治作用及其部分机制，及对肝组织bax和bcl-2基因表达的影响。方法以四氯化碳（CCl4）诱导形成大鼠肝实质损伤性肝纤维化模型。大鼠分为空白对照组、模型对照组、预防组及治疗组。除空白对照组外，其余各组大鼠均皮下注射40％CCl4橄榄油混合液，每3日1次，共10周。预防组同时给予Ena灌胃。治疗组则在造模第5周给予Ena灌胃。对肝组织标本炎症及纤维化程度进行评价，并用RT—PCR技术观察Ena对肝组织bax和bcl-2基因表达的影响。结果Ena高剂量预防及高剂量治疗组肝组织炎症和纤维化程度较模型对照组显著减轻（P〈0．01）。Ena高剂量预防及高剂量治疗组肝组织bax基因表达显著弱于模型对照组（P〈0．01），bcl-2基因表达显著强于模型对照组（P〈0．01），bax／bcl-2基因表达比值和肝纤维化程度呈显著直线正相关（P〈0．01）。结论Ena能有效防治大鼠肝纤维化，抑制促凋亡基因bax的表达，促进抑凋亡基因bcl-2的表达，是其可能机制之一。bax／bcl-2基因表达比值与肝纤维化程度正相关。     

食管鳞癌中p53与bcl-2的表达及其临床意义

目的 :探讨食管鳞癌中p53和bcl 2的表达情况、相互关系及其临床意义。方法 :应用免疫组化法 (S P)对 1 0 0例食管鳞癌病人的内镜活检标本检测p53和bcl 2的表达。结果 :在癌旁正常鳞状上皮、癌旁不典型增生和癌组织中p53的阳性表达率分别为8.9%、43.6 %和 58.6 % (χ2 =30 .2 ,P <0 .0 1 ) ;bcl 2阳性表达率分别为 6.7%、3 .6 %和 43.0 % (χ2 =57.4,P <0 .0 1 )。p53阳性表达率在不典型增生中已接近癌组织 ,bcl 2在癌旁正常鳞状上皮和不典型增生组织中表达较低 ,在癌组织中表达较高。在分化较好的食管鳞癌中 ,p53的阳性表达率和表达强度等级明显高于bcl 2 ,在低分化癌中 ,p53和bcl 2的阳性表达率和表达强度等级无差异。结论 :p53表达可能是食管鳞癌的早期事件 ,bcl 2表达较晚 ,是癌变的特征。p53和bcl 2表达的相互关系与食管鳞癌的分化程度有关     

bcl-2蛋白在H460和H446中的表达及其应激变化

培养正常肺细胞系MRC-5和两株肺肿瘤细胞系H460、H446,应用Western blot法检测bcl-2蛋白的表达;采用120 mj/cm2剂量紫外线(UV)照射,继续培养并对bcl-2蛋白进行免疫组化观察.Western blot结果示,在肺肿瘤细胞系H460和H446中的bcl-2蛋白表达明显高于正常细胞系MRC-5;免疫组化检测示,UV照射后培养10.5 h,bcl-2的表达在3种细胞系中均显著高于未照射组.认为bcl-2蛋白过表达与肺肿瘤的发生有关,但肺肿瘤细胞中bcl-2蛋白的过表达未影响其对UV照射引起的应激反应,从而使肿瘤细胞具有了在应激条件下较正常细胞更强的生存能力.     

Bcl-2及相关蛋白bax在人肝胆管癌中的表达及意义

应用免疫组化方法研究了２１例肝胆管癌中ｂｃｌ－２及相关蛋白ｂａｘ的表达，以探讨两者与肝胆管癌发生的关系。结果发现２１例肝胆管癌细胞阳笥１５例，阳性率为７１．４％，ｂｃｌ－１表达与组织学分级有明显关系。     

凋亡相关基因caspase-9,bax及bcl-2在大肠癌中的表达及其意义

目的探讨凋亡相关基因easpase-9,bax和bel-2在大肠癌中的表达及其在大肠癌发生、发展中的可能作用及相互关系。方法应用免疫组化S-P法检测20例正常大肠黏膜、48例大肠腺瘤及56例大肠癌中的caspase-9、bax和bcl-2蛋白的表达。用TUNEL 法检测细胞凋亡。结果正常大肠黏膜、大肠腺瘤和大肠癌中caspase-9的阳性表达率分别为5．00％、33．33％、64．29％,其表达率在三者间差异有显著性(P<0．01)。bax在三种组织中的表达率分别为5．00％、35．42％、62．50％,三者间差异有显著性(P<0．01)。bcl-2 在三者中的阳性表达率分别为15．00％、87．50％、60．71％,三者间差异亦有显著性(P<0．01)。caspase-9,bax和bcl-2的表达与大肠癌的分化程度有关(P<0．01),与Dukes分期无关(P>05)。正常大肠黏膜、腺瘤和大肠癌中细胞凋亡指数差异有显著性(P<0．01),细胞凋亡指数与肿瘤的分化程度有关(P<0．01),与Dukes分期无关(P>0．05)。caspase-9、bax和bcl-2的表达与细胞凋亡指数有密切联系 (P<0．01)。结论肿瘤早期阶段的细胞凋亡异常,可能是大肠癌的发病原因之一。bcl-2和bax通过调节caspase-9参与大肠癌的发生。     

Relationship between expression and distribution of cyclooxygenase-2 and bcl-2 in human gastric adenocarcinoma

AIM: To explore expression and distribution features of COX-2 and bcl-2 in human gastric adenocarcinoma tissues and to study its biological significance. METHODS: Totally 36 human gastric carcinoma samples were enrolled in this study (cardiac adenocarcinoma 16 cases, distal gastric adenocarcinoma 20 cases). The expressions of COX-2 and bcl-2 in cancerous tissues and corresponding para-cancerous tissues were investigated by immunohistochemistry using COX-2 polyclonal antibody and bcl-2 monoclonal antibody. The normal gastric mucosa tissues were used as control. RESULTS: The expressions of COX-2 and bcl-2 in gastric carcinoma were significantly higher than that in the para-cancerous tissues (77.8% vs 47.2%, P<0.01, 80.56% vs 58.33%, P<0.05). The expression of COX-2 in cardiac adenocarcinoma was remarkably higher than that in the distal gastric carcinoma (93.8% vs 65.0%, P<0.01). The expression of COX-2 was mainly localized in the cytoplasm of tumor cells and partly in the nucleus. There is a transition of the COX-2 cytoplasmic positivity to nucleic in tumor cells with the increase of gastric carcinoma pathological grade. Interstitial macrophages, fibroblasts and vascular endothelial cells also expressed COX-2. The tissues with higher expression of COX-2 also expressed high level of bcl-2 protein. CONCLUSION: Abnormal expression pattern of COX-2 within the tissues of human gastric cancer is correlated with tumor location and lymph node metastasis. COX-2 may regulate expression of apoptosis suppressor gene (bcl-2) through interaction of tumor cells and stromal cells and play an important role in the generation and development of tumors, which will be of great help in developing new methods for antitumor therapy.     

卡维地洛（Carvedilol）对心肌bcl-2、bax表达的影响

目的　研究卡维地洛对心肌凋亡相关基因bcl 2、bax表达的影响。方法　SD大鼠随机分三组 :对照组 ,阿霉素组 ,阿霉素 卡维地洛组。阿霉素腹腔注射 (2 0mg/kg)制备心肌毒性动物模型 ;卡维地洛组在注射阿霉素之后立即腹腔注射卡维地洛 (2mg/kg) ;等量生理盐水用于对照组。于用药后第 1,3,5 ,10 ,15d采用逆转录聚合酶链反应 (RT PCR)和免疫组织化学染色法检测心肌bcl 2、bax基因和蛋白水平表达的变化。结果　 (1)以GAPDH为内标 ,第 1、3、5、10d ,阿霉素组中bcl 2基因表达逐渐减弱 ,bax基因表达逐渐增强 ;阿霉素 卡维地洛组中bcl 2表达逐渐增强 ,于第 5、10天bcl 2表达高于阿霉素组 ,bax表达逐渐减弱 ,于第 5、10天bax表达低于阿霉素组 ;(2 )阿霉素 卡维地洛组Bcl 2、Bax阳性心肌细胞百分比较阿霉素组均有显著性差异 (P <0 0 5 )。结论　 (1)阿霉素心肌毒性与促进bax的表达同时抑制bcl\|2表达有关 ;(2 )卡维地洛可以促进bcl 2的表达 ,减弱bax的表达。提示卡维地洛的心肌保护作用可能通过差异调节bcl 2和bax的表达而实现。     

Effect of bax,bcl—2 and bcl—xL on regulating apoptosis in tissues of normal liver and hepatocellular carcinoma

AIM: To investigate the expression of bax, bcl-2 and bcl-xL mRNA in the tissues of normal liver and hepatocellular carcinoma (HCC), and analyze the relationship between the expression of bax, bcl-2 and bcl-xL mRNA and clinical parameters of HCC patients. METHODS: The expression of bax, bcl-2 and bcl-xL mRNA of normal liver and HCC was measured by Northern blot. Statistical analyses were made by t test and correlation analysis. RESULTS: A very low mRNA level was indicated at bax, bcl-2 and bcl-xL in the HCC tissues in contrast to the tissues of normal liver by Northern blot analysis. The analyses of mRNA level revealed that HCC tissues exhibited a mean 7.6-fold decrease in bax, 4.2-fold in bcl-2 and 3.5-fold in bcl-xL in comparison with normal control tissues, respectively. Positive correlation was found between bax and bcl-xL (r=0.7061, P<0.01). There was no significance between the mRNA expression of these three genes and age, gender, tumor differentiation and tumor stage of HCC patients. CONCLUSION: The results are consistent with the fact that apoptosis rarely occurs in normal livers but increases in HCC, indicating that bcl-2 and bcl-xL may play a very important role in regulating the apoptosis of normal liver and HCC.     

凋亡抑制基因survivin bcl-2 bax在肺癌中表达的研究

目的检测肺癌患者癌组织和癌旁组织中survivin、bcl-2及bax的基因表达,探讨它们的相关性及与肺癌发生、发展的关系。方法应用TUNEL原位细胞凋亡检测方法及免疫组化方法,对1998—2004年华中科技大学同济医学院附属协和医院收治的163例肺癌患者手术常规石蜡包埋组织中癌基因survivin、bcl-2及bax的表达进行检测,并与其中106例患者的癌旁组织对比,分析免疫组化结果及其与肺癌的病理特征和预后关系。结果在癌旁肺组织中,survivin、bcl-2、bax蛋白阳性表达率分别为1·9%(2例)、29·2%(31例)、93·47%(99例);肺癌组织内,三者阳性表达率分别为69·9%(114例)、62·0%(101例)、52·8%(86例)。异常增高的survivin、bcl-2表达呈明显相关。结论细胞凋亡和增殖失控,相关基因survivin、bcl-2和bax蛋白异常表达在肺癌发生发展中起重要作用。survivin、bcl-2可能在肺癌癌变及浸润过程中起着重要作用,可作为判断肺癌生物学行为和预后的参考指标。     

Correlation between expression of gastrin, somatostatin and cell apoptosis regulation gene bcl-2／bax in large intestine carcinoma

AIM: To explore the correlation between expression of somatostatin (SS), gastrin (GAS) and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma. METHODS: Sixty-two large intestine cancer tissue samples were randomly and retrospectively selected from patients with large intestine carcinoma. Immunohistochemical staining for bcl-2, bax, GAS, SS was performed according to the standard streptavidin-biotin-peroxidase (S-P) method. According to the semi-quantitative integral evaluation, SS and GAS were divided into three groups as follows. Scores 1-3 were defined as the low expression group, 4-8 as the intermediate expression group, 9-16 as the high expression group. Bax and bcl-2 protein expressions in different GAS and SS expression groups of large intestine carcinoma were assessed. RESULTS: The positive expression rate of bax had a prominent difference between SS and GAS high, intermediate and low expression groups (P<0.05, x2SS = 9.246; P<0.05, x2GAs = 6.981). The positive expression rate of bax in SS high (80.0%, 8/10) and intermediate (76.5%, 13/17) expression groups was higher than that in low expression group (40.0%, 14/35) (P<0.05, X2high vslow = 5.242; P<0.05, x2middle vs low = 6.097). The positive expression rate of bax in GAS high expression group (27.3%, 3/8) was lower than that in low expression group (69.4%, 25/36) (P<0.05, x2 = 4.594). However, bax expression in GAS intermediate expression group (46.7%, 7/15) was lower than that in low expression group, but not statistically significant. The positive expression rate of bcl-2 had a prominent difference between SS and GAS high, intermediate and low expression groups (P<0.05, x2ss = 7.178; P<0.05, x2GAS = 13.831). The positive expression rate of bcl-2 in GAS high (90.9%, 10/11) and intermediate (86.7%, 13/15) expression groups was higher than that in low expression group (44.4%, 16/36) (P<0.05, x2high,vslow = 5.600; P<0.05, x2middle vs low = 7.695). However, the positive expression rate of bcl-2 in SS high (40. 0%, 4/10) and intermediate (47.1%, 8/9) expression groups was lower than that in low expression group (77.1%, 27/35) (P<0.05, x2high vs low = 4.710; P<0.05, x2middle vs low = 4.706). There was a significant positive correlation between the integral ratio of GAS to SS and the integral of bcl-2 (P<0.01, r = 0.340). However, there was a negative correlation between the integral ratio of GAS to the SS and bax the integral of (P<0.05, r= -0.299). CONCLUSION: The regulation and control of gastrin, somatostatin in cell apoptosis of large intestine carcinoma may be directly related to the abnormal expression of bcl-2, bax.     

叶酸对胃癌前病变bcl-2、bax及p53 基因的影响

目的研究叶酸治疗对胃癌前病变组织中bcl-2、bax及p53基因表达的影响.方法胃镜活检经病理证实为胃癌前病变患者38例,利用逆转录聚合酶链式反应(RT-PCR)方法检测胃癌前病变组织bcl-2、bax基因表达率,利用流式细胞仪检测组织p53蛋白表达率.将患者随机分为治疗组(叶酸10mg,每天三次)与对照组(硫糖铝1.0,每天四次)各19例,治疗结束复查组织中bcl-2,bax基因表达率及p53蛋白表达率.结果治疗组治疗后bcl-2基因表达率降低(P<0.05),bax基因表达率无明显变化(P>0.05),p53蛋白表达率增高(P<0.05),对照组治疗后各项指标无明显变化(P>0.05).结论叶酸干预可促进胃癌前病变组织中p53基因的表达,抑制bcl-2基因的表达,而对bax基因表达无明显影响.     

苦参素注射液联合顺铂对人肝癌SMMC-7721细胞凋亡相关基因c-myc、bcl-2和bax表达的影响

目的探讨苦参素注射液(MI)联合顺铂(DDP)对人肝癌SMMC-7721细胞凋亡相关基因c-myc、bcl-2和bax表达的影响。方法分别采用MI、顺铂及MI联合顺铂干预SMMC-7721细胞。TUNEL法检测细胞凋亡率,半定量RT-PCR法检测c-myc、bcl-2和bax mRNA表达,二步法免疫组化检测c-myc、bcl-2和bax蛋白表达。结果苦参素组、顺铂组和联合用药组细胞凋亡率显著上升,与对照组(不干预)比较差异有统计学意义(P<0.05或P<0.01),联合用药具有单纯相加或协同作用;联合用药组的细胞凋亡率显著增加,bax mRNA和蛋白表达显著升高,c-myc、bcl-2 mRNA和蛋白表达显著减少,与DDP单药组比较差异有统计学意义(P<0.05或P<0.01)。结论苦参素注射液联合顺铂具有单纯相加或协同诱导肝癌SMMC-7721细胞凋亡作用,其机制可能与bax基因表达上调和c-myc、bcl-2基因表达下调有关。     

自身免疫性甲状腺疾病甲状腺组织中bcl-2家族蛋白的表达

目的研究凋亡相关基因bcl鄄2家族蛋白bcl鄄2、mcl鄄1、bcl鄄XL和bax在自身免疫性甲状腺疾病(AITD)甲状腺组织中的表达特征及与AITD发病机制之间的内在联系。方法以甲状腺腺瘤旁正常甲状腺组织为对照(C组,20例),采用免疫组织化学ElivisionTM二步染色法检测凋亡相关蛋白bcl鄄2、mcl鄄1、bcl鄄XL和bax在桥本甲状腺炎(HT组,33例)和Graves病(GD组,28例)患者甲状腺组织中的表达与分布。结果bcl鄄2蛋白表达强度GD组>C组>HT组(P<0.01);mcl鄄1蛋白表达强度GD组>C组>HT组(P<0.01);bcl鄄XL蛋白表达强度HT组和GD组强于C组(P<0.01),但HT组和GD组间差异无统计学意义(P>0.05);bax蛋白的表达强度HT组>GD组和C组(P<0.01),但GD组和C组间差异无统计学意义(P>0.05);HT组中,在淋巴细胞浸润区域附近的甲状腺滤泡上皮细胞(TEC)bcl鄄2表达弱,bax和mcl鄄1表达强;远离淋巴细胞浸润区域的TECbcl鄄2表达强,bax和mcl鄄1表达弱(P<0.05)。结论(1)抗凋亡bcl鄄2和mcl鄄1蛋白在HT中表达的减弱以及在GD中表达的增强对于HT甲状腺滤泡细胞凋亡的增加和GD甲状腺滤泡细胞的增殖可能起一定作用;(2)bax蛋白在HT中表达增强所起的促凋亡的作用对疾病的发生发展起一定作用;(3)bcl鄄2与bax表达强度的比值对于凋亡的调控起重要作用;(4)bcl鄄2家族蛋白bc