多囊肾病囊肿衬里上皮细胞增生与凋亡的研究

目的 :研究多囊肾病囊肿衬里上皮的增生与凋亡及相关蛋白表达。　 方法 :应用原位末端标记(TUNEL)法和免疫组化法 ,对 3例正常肾组织和 12例常染色体显性遗传型多囊肾病 (ADPKD)患者 (2例为终末期肾病患者 ,10例为氮质血症期患者 )肾组织中囊肿衬里上皮细胞的凋亡细胞和增生细胞核抗原 (PCNA)阳性细胞计数 ,并检测c myc、bcl 2、bax、p5 3、Fas蛋白表达。　 结果 :多囊肾病囊肿衬里上皮细胞增生与凋亡均增加 ,增生较凋亡增加显著 ,PCNA阳性细胞 /凋亡细胞为 3 46± 1 6 8。多囊肾组织c myc蛋白和bcl 2蛋白表达显著升高。bax、p5 3和Fas的表达并无改变。　 结论 :多囊肾病中囊肿衬里上皮细胞的增生和凋亡失调 ,调节这些过程的基因异常表达介导了多囊肾病囊肿形成。多囊肾组织c myc蛋白高度表达 ,与囊肿衬里上皮过度增生、凋亡相关。多囊肾囊肿衬里上皮细胞的凋亡不依赖于bcl 2、p5 3和Fas。     

缺血预适应减少心肌缺血损伤机制的研究

目的　通过对心肌缺血预适应的动物模型观察 ,探讨细胞凋亡在其中的作用 ,以及p5 3,bcl 2 ,Bax基因对其发生进行的调控。方法　采用TUNEL标记技术研究心肌缺血预适应心肌细胞中细胞凋亡现象 ,并采用免疫组化染色技术及原位分子杂交技术研究p5 3,bcl 2及Bax基因的蛋白及mRNA的表达。结果　缺血预适应组 (P)及非缺血预适应组 (NP)非缺血区均未见凋亡细胞 ,但在P组缺血区可见散在的凋亡细胞 ,而在NP组缺血区则多见。P组p5 3蛋白表达显著低于NP组 ,bcl 2蛋白表达在P组显著高于NP组 ,Bax蛋白表达在P组显著低于NP组 ,并且bcl 2 /Bax的比值P组与NP组相比显著升高。P组p5 3基因mRNA表达显著低于NP组 ,bcl 2基因mRNA表达在P组显著高于NP组。结论　心肌缺血预适应对心肌的保护可通过抑制细胞凋亡来实现 ,并且通过bcl 2表达增加 ,p5 3、Bax表达减少对其进行调控。     

吲哚美辛诱导胃癌细胞凋亡的机制研究

目的 :研究非甾体类抗炎药吲哚美辛诱导人胃癌细胞系SGC 790 1的凋亡作用及对COX 2mRNA表达及c myc、bcl 2、cas pase 3凋亡基因蛋白的表达 ,以探索其凋亡机制。方法 :胃癌细胞的凋亡用电子显微镜、AnnexinV FITC染色流式细胞仪技术测定。COX 2基因表达用RT PCR法测定。c myc、bcl 2和caspase 3蛋白表达用免疫细胞化学技术测定。结果 :吲哚美辛在浓度为 50 μmol/L时作用48、72h和浓度为 1 0 0和 2 0 0 μmol/L时作用 2 4、48、72h均可诱导胃癌细胞凋亡 ,凋亡率分别为 6 .48%、8.2 0 % ;9.1 4 %、1 2 .2 7%、1 5 .1 1 %和 9.95 %、1 4 .70 %、1 9.81 % ,呈浓度和时间依赖性。吲哚美辛可降低COX 2mRNA表达和bcl 2蛋白表达 ,增加c myc和caspase 3蛋白表达。 2 0 0 μmol/L吲哚美辛作用 72h ,bcl 2、c myc和caspase 3蛋白表达阳性率分别为 2 2 .8± 6 .5 %、42 .5± 1 3 .1 %和 31 8± 1 2 .7% ;对照组为 44 .6± 1 0 .1 %、2 4 .74± 9.5 %和 1 4 .8± 6 .4%。两者比较 ,差异有显著性 (P <0 .0 1 )。结论 :吲哚美辛可诱导胃癌细胞SGC 790 1凋亡 ,其机制涉及bcl 2表达下调、c myc表达上调及caspase 3激活。说明多基因调控参与其中     

慢性心力衰竭兔模型心肌细胞凋亡及其基因调控的实验研究

目的 :研究持续快速心室起搏致慢性心力衰竭 (心衰 )兔模型心肌组织的细胞凋亡 ,以及基因 p5 3、bcl 2、Bax在心肌细胞中的表达。方法 :采用持续快速心室起搏的方法建立慢性心衰兔模型为心脏起搏组 (n =7) ,运用TUNEL标记法检测兔模型心肌组织中的凋亡细胞 ,并运用免疫组化及原位分子杂交技术研究基因 p5 3、bcl 2、Bax在心肌细胞中的表达 ,并与假手术组 (n =9)进行对比。结果 :在心脏起搏组约每 10 6个心肌细胞中有 40 33± 884 5 6个凋亡细胞 ,而假手术组则无 ;心脏起搏组中P5 3蛋白表达、Bax蛋白表达显著高于假手术组 ,而bcl 2蛋白表达则明显低于假手术组。心脏起搏组核糖核酸在心肌细胞中的表达与假手术组比较 ,P5 3明显升高 ,bcl 2则明显降低 ,均有非常显著性差异 (P均 <0 0 1)。结论 :细胞凋亡参与了持续快速心室起搏致慢性心衰的发生及发展 ,并且基因 p5 3的激活、blc 2的表达抑制、Bax的表达增加 ,具有促其细胞凋亡的作用。     

白血病细胞p53和bcl-2表达与化疗药物诱导凋亡的关系

目的　探索p5 3和bcl 2表达及化疗药物体外诱导白血病细胞凋亡与患者化疗疗效的关系。方法　用免疫组化法测定 42例急性白血病患者骨髓细胞p5 3和bcl 2表达水平 ;用TdT介导的脱氧核苷酸切口和末端标记法 (Tunel法 )、Wright Giemsa染色及超微结构观察等方法检测化疗药物诱导的白血病细胞凋亡。结果　 42例急性白血病患者p5 3和bcl 2总体表达水平显著高于对照组。2 7例随访组患者完全缓解 (CR) 16例 ,未缓解 (NR) 11例。CR者表现以p5 3和bcl 2共同阴性及化疗诱导高凋亡为主 ,NR者以p5 3和bcl 2共同阳性及化疗诱导低凋亡占优势。形态学观察可见典型细胞凋亡特征。结论　检测急性白血病患者p5 3和bcl 2表达水平及体外化疗药物诱导白血病细胞凋亡的敏感性有助于预测患者的化疗效果。     

bcl-2,c-myc蛋白在胃癌前病变、胃癌中的表达

目的观察bcl－2，c－myc蛋白在胃癌前病变、胃癌中的表达．方法取内镜活检标本93例，其中慢性浅表性胃炎（CSG）21例，伴有上皮异型增生（gastricepithelialdysplasia，GED）者5例．慢性萎缩性胃炎（CAG）34例，伴有GED、肠上皮化生（IM）者23例．胃癌（GC）38例，癌旁粘膜伴有GED和IM者22例．SP免疫组化法检测bel－2，c－myc基因蛋白．结果bcl－2基因蛋白弥漫分布于细胞浆中，c－myc基因蛋白表达多在细胞浆中，少数细胞核内同时阳性．bcl－2，c－myc基因蛋白表达在CSG伴有GED粘膜中的阳性率分别为100％和60．0％．不伴GED组，前者为零，后者为75．0％在CAG伴有GED和IM粘膜中bcl－2，C－myc的阳性率分别为34．8％和73．9％．无GED和IM粘膜则前者为零，后者为63．6％．在胃炎中，伴有GED，IM与不伴有GED，IM组比较，bcl－2表达有显著性差异P＜0．05—0．001．c－myc表达则差异不明显P＞0．05．在GC中，bcl－2，c－myc阳性表达率分别为47．4％和76．3％．在GC癌旁粘膜中，伴有GED和IM者bcl－2阳性率为54．5％，c－myc为81．8％．不伴有GED，IM者bcl-2无阳性表达，c－myc阳性率为25％．GC及伴有GED，IM癌旁粘膜与不伴GED，IM的癌旁粘膜比较，bcl－2与c－myc的表达差异均非常显著（P＜0．001）．bcl－2，c－     

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     

高浓度葡萄糖对人胰岛β细胞凋亡影响的分子机制

目的　初步探讨高浓度葡萄糖对人胰岛 β细胞凋亡的影响及其分子机制。　 方法　分离培养人胰岛细胞 ,并分为对照组、高糖组和高糖 氨基胍组 ,3 7℃ ,5 %CO2 培养 72h ,测定培养液上清液中胰岛素、一氧化氮 (NO)、还原性谷胱甘肽 (GSH)水平。原位末端核苷酸标记法 (TUNEL)和胰岛素免疫组化双染色法及ELISA法检测胰岛 β细胞凋亡 ,RT PCR检测胰岛细胞p5 3、Bcl 2和胰岛素基因启动转录因子 1 (PDX 1 )mRNA表达水平。　结果　高糖组胰岛 β细胞凋亡小体富计因子(1 91± 0 6 9)、β细胞凋亡率 (1 4 8% )、NO〔(1 82 3± 1 5 5 ) μmol/L〕和 p5 3mRNA(0 3 0 6± 0 0 3 9)表达水平均显著高于高糖 氨基胍组〔分别为 1 1 9± 0 3 3、6 8%、(1 5 4 2± 1 9 7) μmol/L、0 1 3 9±0 0 6 9,P <0 0 1〕和对照组〔分别为 1 0 6± 0 2 6、4 2 %、(1 1 7 3± 2 1 7) μmol/L、0 1 2 5± 0 0 1 5 ,P <0 0 5〕 ,而胰岛素释放量、GSH、bcl 2mRNA和PDX 1mRNA表达水平则显著低于高糖 氨基胍组(P <0 0 5 )和对照组 (P <0 0 5 )。　结论　高浓度葡萄糖可通过诱导人胰岛 β细胞凋亡及PDX 1表达降低使胰岛素分泌减少 ,其机制与高糖状态下胰岛 β细胞抗氧化能力降低引起NO介导的p5 3高表达和PDX 1低表     

舒心胶囊对缺氧诱导人脐静脉内皮细胞凋亡及相关基因表达的影响

观察舒心胶囊对缺氧诱导人脐静脉内皮细胞凋亡及相关基因表达的影响。体外培养人脐静脉内皮细胞 ,建立缺氧模型 ,以透射电子显微镜技术、流式细胞术 (膜联蛋白v/碘化丙啶 )双染色鉴定细胞凋亡 ;流式细胞术检测Fas、bcl 2及p5 3蛋白表达。缺氧培养 12h后 ,电镜显示细胞呈典型凋亡形态 ,双染色流式细胞仪检测凋亡细胞比例 (13.0 3%± 0 .96 % )显著高于正常组 (4 .4 7%± 0 .6 4 % ) (P <0 .0 1) ,而加入 5g/L舒心胶囊原液缺氧培养 12h后 ,凋亡细胞显著减少 (5 .0 3%± 0 .5 4 % ) (P <0 .0 1)。Fas蛋白表达在缺氧损伤过程中未见改变 ,舒心胶囊可使bcl 2蛋白表达升高 ,p5 3蛋白表达降低。舒心胶囊能够抑制缺氧诱导人脐静脉内皮细胞凋亡 ,可能与其升高bcl 2表达、降低p5 3表达有关。     

下调肌细胞增强因子2C基因对动脉粥样硬化血管内皮细胞增殖凋亡及核因子-κB信号通路的影响

目的探讨下调肌细胞增强因子(MEF) 2C基因对动脉粥样硬化(AS)血管内皮细胞增殖凋亡及核因子(NF)-κB信号通路的影响及机制。方法 Western印迹检测AS患者斑块组织和正常对照组MEF2C的蛋白表达;将MEF2C的特异性siRNA(si-MEF2C)及阴性对照siRNA(NC)转染人脐静脉血管内皮(ECV)-304细胞,Western印迹检测转染效果。ECV-304细胞经H_2O_2处理后转染si-MEF2C,细胞分为NC组、H_2O_2组、H_2O_2+si-MEF2C组和正常对照组,噻唑蓝(MTT)检测细胞活力;流式细胞术检测细胞凋亡率及活性氧(ROS)水平; Western印迹检测核因子(NF)-κB p65、NFkappa B激酶抑制剂(IKK)α、增殖细胞核抗原(PCNA)和Bcl-2的蛋白表达。结果 AS患者斑块组织MEF2C蛋白表达显著高于正常对照组(P<0. 05); MEF2C的特异性siRNA转染ECV-304细胞后MEF2C蛋白表达显著低于空白对照组(P<0. 05)。H_2O_2组细胞活力及PCNA和Bcl-2的蛋白表达均显著低于NC组,细胞凋亡率、ROS水平及NF-κB p65和IKKα的蛋白表达均显著高于NC组(P<0. 05),而H_2O_2+si-MEF2C组细胞活力及PCNA和Bcl-2的蛋白表达均显著高于H_2O_2组,细胞凋亡率、ROS水平及NF-κB p65和IKKα的蛋白表达均显著低于H_2O_2组(P<0. 05)。结论下调AS血管内皮细胞MEF2C基因表达可提高细胞活力,抑制细胞凋亡,机制可能是细胞内ROS水平降低及NF-κB信号通路的下调。     

动脉粥样硬化斑块中平滑肌细胞凋亡及凋亡相关基因的表达

为了揭示动脉粥样硬化发生发展过程中血管平滑肌细胞增殖与凋亡的关系及其调节机制，采用末端脱氧核糖核酸转移酶介导的dUTP缺口末端标记（TUNEL）染色法对家兔动脉粥样硬化斑块组织中平滑肌细胞的凋亡情况进行原位检测，并对原癌基因bcl-2、c-myc和P53在斑块组织中的表达活性进行North－ernblot分析。结果发现，在动脉粥样硬化斑块组织中有许多TUNEL染色阳性的平滑肌细胞，提示斑块组织中有平滑肌细胞凋亡发生：其斑块组织的DNA电泳图谱呈梯形状．符合细胞凋亡的特征性改变。Northern印迹分析表明，在动脉粥样硬化斑块组织中，原癌基因bck-2和P53的表达较正常组织增强，C－myc表达减低。提示这些基因的表达对动脉粥样硬化斑块组织中的平滑肌细胞凋亡起着重要的调节作用。     

The function of apoptosis and protein expression of bcl-2, p53 and C-myc inthe development of gastric cancer

AIM To understand the rule and possible function of apoptosis and protein expression of bcl-2, p53 and C-myc in chronic gastritis, gastric ulcer, non-classic proliferation of gastric mucosa and gastric cancer.METHODS Apoptosis was detected by using in situ terminal labelling (TUNEL). The protein expression ofbcl-2, p53 and C-myc was detected by immunohistochemical method.RESULTS The indexes of apoptosis in chronic active gastritis, gastric ulcer, mild and severe non-classicproliferation of gastric mucosa, early and progressive gastric cancer were 16.8%±12.3%, 24.1%±20.0%,19.3%±16.4%, 15.7%±15.2%, 10.1%±9.1% and 6.3%±6.0%, respectively. The index of progressivegastric cancer was lower than that of early gastric cancer and non-classic proliferation of gastric mucosa(P<0.05). The positive rate of bcl-2 protein was 9.4%, 27.6%, 52.9%, 75.0%, 83.3% and 46.7%,respectively. The positive rate of bcl-2 of early gastric cancer was higher than that of progressive gastriccancer. The positive rates of p53 protein of severe non-classic proliferation, early and progressive gastriccancer were 25.0%, 33.3% and 63.3%, respectively. The positive rate of p53 of progressive gastric cancerwas higher than that of early gastric cancer and non-classic proliferation (P<0.05). In Lauren types, theindex of apoptosis, protein expression rates of bcl-2, p53 and C-myc of intestinal type were 8.3%±7.2%,38.9%, 77.7% and 56.6%, while that of diffuse type were 5.1%±4.9%, 58.3%, 50.0% and 8.3%,respectively. All markers had statistical difference between two types (P<0.05).CONCLUSION Apoptosis was inhibited stepwise in the development of non-classic proliferation of gastricmucosa to early gastric cancer and then to progressive gastric cancer. The high expression of bcl-2, p53 andC-myc was related to the development of gastric cancer, bcl-2 might play an important role in early gastriccancer while p53 and C-myc act mostly in middle and late stage gastric cancer. The Lauren typing of gastriccancer is closely related to the index of apoptosis and expression of bcl-2, p53 and C-myc.     

Dual effects of 8-Br-cAMP on differentiation and apoptosis of human esophageal cancer cell line Eca-109

AIM: To investigate the effects of 8-Br-cAMP on differentiation and apoptosis of human esophageal cancer cell line Eca-109, and the related gene expression. METHODS: The cultured Eca-109 cells were divided into four groups: E1 group (co-cultured with 8-Br-cAMP for 24 h); E2 group (co-cultured with 8-Br-cAMP for 48 h); C1 group (treated without 8-Br-cAMP for 24 h); and C2 group (treated without 8-Br-cAMP for 48 h). The same concentration of cell suspension of each group was dropped separately onto the slides and nitrocellulose membranes (NCM). The biotin-labeled cDNA probes for c-myc, wild-type (wt) p53, bcl-2 and iNOS were prepared for in situ hybridization. The expressions of epidermal growth factor receptor (EGFR), p38 kinase, FAS, FasL and caspase-3 were detected using immunocytochemistry, and the NOS activity and the ratio of differentiated cells/proliferating cells were examined by cytochemistry. Immunocytochemistry, cytochemistry, and in situ hybridization were separately carried out on both slides and NCM specimens for each group. In addition, TUNEL was used to detect the cell apoptosis rate in each group. RESULTS: The apoptotic rate of E2 group was significantly higher compared to E1 group, while there was no difference in the ratio of differentiated cells/ proliferating cells between E1 and E2 groups. The signals of wt p53 and iNOS were markedly stronger, while the signals of c-myc and EGFR were obviously weaker in E1 group than those in C1 group (P<0.05). Moreover, the signals of wt p53, iNOS, p38 kinase, caspase-3 and NOS activity were significantly stronger, whereas, the signals of bcl-2, c-myc and Fas/FasL were markedly weaker in E2 group than those in C2 group (P<0.05). CONCLUSION: The differentiation and apoptosis of human esophageal cancer cell Eca-109 can be induced after 24- and 48-h treatment with 8-Br-cAMP, respectively. Upregulation of wt p53, iNOS and downregulation of c-myc may be associated with differentiation and apoptosis of Eca-109 cells. Furthermore, upregulation of FasL, p38 kinase and caspase-3 as well as downregulation of bcl-2, and Fas may be involved in the apoptosis of Eca-109 cells.     

X-ray induced L02 cells damage rescued by new anti-oxidant NADH

AIM：To explore molecular mechanism of nicotinamide adenine dinucleotide (NADH) antagonization against X-ray induced L02 cells damage.METHODS: L02 liver cells were cultured in RPMI 1640,exposed to X-ray irradiation and continued to culture in the presence or absence of NADH. Cellular viability was analyzed by routine MTT methods. The percent age of apoptotic cells and positive expressions of pS3, bax and bcl-2, fas, fasL proteins were determined by FCM. Level of intracellular ROS was determined by confocal microscope scanning. Morphological change was detected by scanning electron micrograph.RESULTS: The viability of L02 cells was decreased with increasing dose of X-ray irradiation. NADH could not only eliminate the apoptosis induced by X-ray irradiation, but also up-regulate expression of bcl-2 protein and down-regulate expression of p53, bax, fas and fasL proteins (P&lt;0.05). At the same time, NADH could reduce level of intracellular ROS in radiated L02 cells.CONCLUSION: NADH has marked anti-radiation effect, its mechanism may be associated with up-regulation of bcl-2 expression and down-regulation of p53, bax fas and fasL expression, as well as decline of intracellular ROS. However,further investigation of its mechanism is worthwhile.     

Dynamic expression of apoptosis-related genes during development of laboratory hepatocellular carcinoma and its relation to apoptosis

AIM: To explore the expression of p53, bcl-2, bax, survivin and the cell apoptosis during the development of tree shrew hepatocellular carcinoma (HCC), the relationship between expression of these genes, its impact on HCC development, and its relation to cell apoptosis. METHODS: Tree shrew HCC was induced with aflatoxin B1 (AFB1), and regular biopsy of liver tissues was carried out and the biopsy tissues were collected during cancer inducement. Liver biopsy tissue and HCC tissue were collected from 35 pre-cancerous experimental animals at wk 30 and 60 and at the 30th-, 60th-, and 90th-wk. Liver biopsy tissues were collected from 13 blank control animals at wk 30, 60, and 90. Expression of p53, bcl-2, bax, and survivin at each stage was examined by immunohistochemistry method. Apoptotic cells were detected in situ by the terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) technique. RESULTS: The apoptosis rate of normal hepatic cells was extremely low, whereas it increased during the formation of HCC. Expression of the apoptosis-related genes p53, bd-2, bax, and survivin during the formation of HCC presented an increasing tendency. Expression of p53 did not noticeably relate to that of bcl-2, bax, and survivin, whereas expression of bcl-2 and bax was closely related. In HCC, p53 did not present a distinct relation to cell apoptosis, whereas its high level expression was probably related to liver cell proliferation. Survivin negatively correlated apoptosis index, and its overexpression could inhibit cell apoptosis. CONCLUSION: Apoptosis-related genes p53, bcl-2, bax, and survivin are all related to the occurrence of HCC. The anti-apoptosis effect of bcl-2 is influenced by bax, and ratio bcl/bax reflects more correctly the extent of cell apoptosis.     

肺癌平药物血清对A549人肺癌细胞凋亡及基因表达的影响

目的探讨肺癌平治疗肺癌的作用机制。方法采用血清药理学方法孵育A549人肺癌细胞，通过电镜、流式细胞术、免疫细胞化学方法检测肺癌平药物血清作用后对细胞凋亡及相关基因bcl-2、p53表达的影响。结果肺癌平药物血清作用后电镜下可见凋亡小体形成，流式细胞仪检测可见亚二倍体核型峰的特征，免疫细胞化学法显示抑癌基因p53主要定位在细胞核，部分细胞浆也有淡染。癌基因bcl-2主要表现为胞浆着色，流式细胞仪分析药物血清作用后，p53表达增强，bcl-2表达降低，随着作用时间延长，两种基因表达明显增强。结论肺癌平治疗肺癌的作用机制是通过诱导肺癌细胞凋亡及影响相关基因bcl-2、p53表达实现的。     

Imbalance between cell proliferation and cellular DNA fragmentation in hepatocellular carcinoma.

AIM/BACKGROUND: Hepatocellular carcinoma (HCC) is known for its rapid growth. This study was undertaken to determine the expression of proliferative markers, apoptosis (DNA fragmentation) and oncogene products known to regulate apoptosis (p53, bcl-2) in HCC. METHODS: 150 Chinese patients with HCC were studied (M:F 128:22, age 14-88 years). Immunohistochemistry was employed to detect cell proliferative markers (PCNA, Ki67), and oncogene products known to regulate apoptosis (p53, bcl-2). DNA fragmentation was determined by terminal dUTP nick end labeling (TUNEL). RESULTS: 98% and 95% of HCC had PCNA (median 2+) and Ki67 (median 2+) detected respectively. TUNEL labeling was detected in only a small number of tumor cells (no labeling in 11%, median 1/1000 cell labeled, range: 0-70/1000 cells). There was no correlation between TUNEL labeling and the clinical parameters (sex, age, cirrhosis, and survival) and the expression of cell proliferative markers. p53 was detected in 53% of the patients (median 1+, range: 0-4+) and bcl-2 was detected in a small proportion of tumor cells in only 13% of the HCCs (range: 0-1 +). The expression of p53 and Bcl-2 did not correlate with TUNEL labeling or the natural survival. CONCLUSIONS: Cell proliferation in HCC is unmatched by apoptosis, accounting for the rapid growth of this tumor. This lack of apoptosis in HCC is unrelated to the expression of p53 or bcl-2 over-expression.     

舒林酸诱导胃癌BGC-823细胞凋亡的研究

目的 探讨舒林酸诱导人胃癌BGC-823细胞株凋亡的作用及其机制。方法将舒林酸作用于人胃癌BGC-823细胞,并设置不同的作用浓度和作用时间,应用流式细胞术、TUNEL法检测胃癌细胞的凋亡率;免疫组化（S-P）法检测凋亡基因蛋白bcl-2、Sru—vivin的表达以及环氧合酶-2（COX-2）蛋白的表达情况。结果 流式细胞仪检测出凋亡峰;其凋亡检测率及TUNEL法检测的细胞凋亡率均高于对照组（P〈0.05）;免疫组化结果显示凋亡基因蛋白bcl-2、survivin等在胃癌细胞表达下降,COX-2蛋白的表达阳性率也显著低于对照组,均呈时间和剂量依赖性（P〈O.05）。结论舒林酸可诱导胃癌BGC-823细胞凋亡,其机制与抑制凋亡抑制基因bcl-2和survivin的表达及下凋COX-2蛋白的表达有关。