Characterization of cell death events induced by anti-neoplastic drugs cisplatin, paclitaxel and 5-fluorouracil on human hepatoma cell lines: Possible mechanisms of cell resistance.

Two different hepatoma cell lines were incubated for 48h with chemotherapeutic drugs cisplatin, paclitaxel and 5-FU to determine their ability to induce cytotoxicity and DNA fragmentation as well as to modify the expression of some cell death-related genes that could be involved in the resistance to therapy. We observed that cisplatin and paclitaxel induced cytotoxicity, but significant differences between both cell lines, were found only in the case of paclitaxel. At 48h, apoptosis was clearly present in Hep3B cells treated with cisplatin and HepG2 cells treated with paclitaxel. 5-FU induced cytotoxicity in both cell lines but only at higher concentrations than the other two drugs, triggering apoptosis and necrosis in HepG2 cells and only necrosis in Hep3B. When a time course was performed for the first 8h of treatment to elucidate the initial mechanism of cell death responsible for DNA fragmentation, we observed that 5-FU in Hep3B, and cisplatin in both cell lines, induces primary necrosis, whereas at the concentration tested here, paclitaxel clearly triggers apoptosis in both cell lines. HepG2 cells were weakly sensitive to 5-FU in the first 8h of treatment, so the primary mechanism of cell death was not clear, but results seem to indicate that it could be apoptosis. At 48h, Bax was not up-regulated with any of the treatments, whereas cisplatin was able to induce Bcl-xL down-regulation in both cell lines. Treatment with 5-FU also down-regulated Bcl-xL in HepG2 cells. We also measured variations in the expression of survivin, an inhibitor of apoptosis that has also been involved in mitototic catastrophe. Hep3B cells seem to show an increase in protein levels with all treatments. Exposure to paclitaxel resulted in the highest effect. In the case of HepG2 cells, there was a decrease in survivin expression when cells were treated with 5FU and paclitaxel, both treatments showing complete loss of the protein. Using an antibody that recognizes unprocessed caspase-3, we observed that the enzyme was assumingly activated in HepG2 cells treated with 5FU and paclitaxel, but only weakly after treatment with cisplatin. Hep3B cells did not show activation since the levels of the pro-enzyme remained the same as that in the control. In conclusion, the three drugs tested in this study could induce cell death, with paclitaxel being more effective inducing apoptosis. 5FU was only effective at high doses and its mechanism seems to be primarily related to necrosis in Hep3B and probably apoptosis in HepG2. Cisplatin mechanism of cell death is probably mediated by the decrease in anti-apoptotic protein Bcl-xL whereas paclitaxel and 5FU are decreasing the apoptosis inhibitor survivin. According to pro-enzyme levels, caspase-3 was only activated in HepG2 cells, whereas in the case of Hep3B cells the mechanisms of toxicity appear to be caspase-3-independent at the time and concentrations tested in this study. The resistance of Hep3B cells to death induced by chemotherapy could be related to an increase in the expression of IAP survivin, which can decrease cell response to the treatment or even switch the type of death from apoptosis to another kind, making therapy less efficient.     

14年间住院病人主要病因和死因变化趋势

目的　探讨 1990～ 2 0 0 3年住院病人的主要病因和死因及变化趋势的方向、强度。方法　分析我院 1990～ 2 0 0 3年录入广东省卫生厅“广东省医院统计病案管理系统”的统计资料 ,计算住院病人主要疾病、死亡病因 1990～ 1999年、2 0 0 0～ 2 0 0 3年两个阶段的年均构成比 ,观察顺位变化 ,用回归分析的方法计算变化趋势。结果　 1) 1990～ 1999年主要住院病因顺位是消化系统疾病 (15 .6 2 % )、损伤和中毒 (13.16 % )、循环系统疾病 (11.2 4 % )、呼吸系统疾病 (10 .4 7% )、恶性肿瘤 (10 .4 3% ) ;2 0 0 0～ 2 0 0 3年顺位是循环系统疾病 (16 .4 1% ) (其中缺血性心脏病占 2 2 .98% ,脑血管病占 39.6 8% ) ,损伤和中毒(12 .80 % )、消化系统疾病 (11.76 % )、呼吸系统疾病 (9.5 3% )、恶性肿瘤 (8.73% ) ,循环系统疾病呈显著上升趋势 (B =4 .2 6 ,P <0 .0 5 ) ,消化系统疾病呈显著下降趋势 (B =- 3.17,P <0 .0 5 ) ;2 ) 1990～ 2 0 0 3年住院死因顺位为恶性肿瘤 (2 9.2 9% )、循环系统疾病 (2 5 .91% )、损伤和中毒 (12 .6 3% )、呼吸系统疾病(6 .18% ) ,消化系统疾病 (5 .77% ) ;2 0 0 0～ 2 0 0 3年恶性肿瘤呈显著下降趋势 (B =- 3.88,P <0 .0 5 ) ,循环系统疾病呈上升趋势但无显著性 (B =0 .84 ,P >0     

食管癌、胃癌组织的低密度脂蛋白受体分析

以１２５Ⅰ－ＬＤＬ为配体对食管癌、胃癌及其相应的正常组织进行了ＬＤＬ受体水平的测定，同时分析了各组织中蛋白质、ＤＮＡ的含量，以探讨食管癌和胃癌发生与ＬＤＬ代谢之间的关系。结果显示，癌组织和正常组织的ＬＤＬ受体亲和性差异无显著性，但癌变组织的蛋白质、ＤＮＡ和ＬＤＬ受体水平均明显高于相应正常组织。癌细胞需要大量的胆固醇满足其有丝分裂，细胞通过受体升高机制使ＬＤＬＲ增多，以增加对ＬＤＬ的摄取和利用。     

胃癌腹腔脱落细胞检查及临床意义

腹膜转移是胃癌常见的转移形式。为在肉眼可见转移前发现亚临床的转移现象,自1981年3月至1982年4月,我们共对胃癌病人142例进行了腹腔脱落细胞检查及临床随诊,发现有腹腔脱落细胞阳性者76例(53.5％)。5年随诊结果发现,腹腔脱落细胞阳性者无一例生存>3年;而腹腔脱落细胞阴性的66例中生存5年者有11例。可见,腹腔脱落细胞检查,对胃癌的诊断,治疗和预后判断有一定价值。     

保留幽门间置空肠双腔代胃术临床研究

目的 探讨全胃切除术后保留幽门间置空肠代胃术的临床应用。方法 1987年至今，对60例胃癌患者临床应用保留幽门的间置空肠代胃术式，评估其疗效。结果 60例患者无手术死亡，术后幽门舒缩功能及代胃的食糜搅拌、贮存功能良好，无返流性食管炎、倾倒综合症、营养不良等并发症。结论 保留幽门的间置空肠双腔代胃术能明显地改善全胃切除术后患者的生活质量。     

功能性消化不良患者胃排空障碍与胃肠激素的关系

目的 探讨功能性消化不良(FD)患者胃排空障碍与胃肠激素间的关系。方法 对54例四患者进行胃排空检查，根据结果将其分为胃排空延缓的FD组(FDD组)和胃排空正常的四组(FDN组)，另以17名正常人作为对照组。用放免法测定受试者血浆(空脂和餐后)、胃窦十二指肠粘膜组织的神经降压素(NT)和胃动宗(MTL)含量。结果 FDD组空脂和餐后血浆、胃窦和十二指肠粘膜组织的NT含量均明显高于对照组及FDN组。各组试餐前后血浆NT增幅差异无显著性。FDD组空脂和餐后血浆、胃窦和十二指肠粘膜组织的MIL含量均明显低于对照组及FDN组。各组十二指肠粘膜组织MTL含量均明显高于胃窦粘膜。结论 FD患者胃排空障碍与NT、MIL密切相关。NT、MIL在FD的发病机制中可能具有一定作用。     

癌胚抗原及粘液组织化学在胃癌中表达的研究

对60例胃癌进行了粘液组织化学和癌胚抗原免疫组织化学研究。结果表明:胃癌发病率肠型和胃肠混合型占多数。60例胃癌中含大肠型粘液者占50例,而单纯含胃型粘液为极少数。癌胚抗原分布方式,肠型胃癌与胃肠混合型胃癌比较无显著差异性(P>0.05)。癌胚抗原的阳性率,胃肠混合型胃癌高于肠型胃癌,两者之间有非常显著差别(P<0.01)。认为选用能显示硫粘蛋白和氧乙酰基唾液酸粘蛋白的粘液组织化学方法,作为胃癌组织分型和确定胃粘膜肠化性质,对癌前病变诊断有一定价值。     

电子计算机监护慢性肝病程序的验证

用参与建立数学模型的521例资料和307例没参与建立数学模型的病例资料验证电子计算机监护慢性肝病的程序。以病理学检查、或以3个月至3年的临床观察结果作为诊断的金标准。254例临床诊断作为对照。验证结果表明（1）计算机监护优于临床诊断，（2）本程序敏感度高。