~(103)钯放射性支架诱导人胆管癌细胞凋亡及对Fas基因表达的影响

目的探讨103钯(Pd)放射性支架释放的γ射线对Fas基因表达的影响以及与胆管癌细胞凋亡的关系及意义。方法建立人胆管癌裸鼠移植瘤模型;将建模成功的裸鼠分为实验组和对照组,实验组置入37MBq103Pd胆道放射性支架,对照组置入普通金属支架;支架置入后10d计算肿瘤体积;应用TUNEL法检测胆管癌细胞凋亡的情况;免疫组化染色方法检测胆管癌细胞中Fas基因表达的情况。结果实验组肿瘤体积较对照组增长明显缓慢(P0.05);胆管癌细胞的Fas基因表达阳性率较对照组明显增高(P0.05),且胆管癌细胞出现明显凋亡,细胞凋亡率明显高于对照组(P0.01)。结论103Pd放射性支架可诱导人胆管癌裸鼠移植瘤中胆管癌细胞凋亡,明显抑制胆管癌细胞生长,其可能通过增强Fas基因表达促进胆管癌细胞凋亡,可能是103Pd放射性支架治疗胆管癌的重要机理之一,进一步为临床应用103Pd放射性支架治疗胆管癌提供了理论依据。     

腺病毒介导凋亡素基因对人胆管癌细胞凋亡的研究

目的探讨凋亡素基因转染对人胆管癌细胞凋亡的影响,为胆管癌的治疗探索新方法。方法采用含凋亡素基因的重组腺病毒感染胆管癌细胞株QBC939,在确定其感染复数后通过光镜、荧光观察其凋亡情况,同时通过TUNEL染色统计凋亡率。结果胆管癌细胞株QBC939的重组腺病毒的感染复数为65,在感染后第3天凋亡素基因能引发胆管癌细胞株QBC939的凋亡,并且随着时间的推移其作用越明显,在第7天凋亡率达到61.9%。结论凋亡素能明显引发人胆管癌细胞的凋亡,在胆管癌的治疗上具有较大潜力。     

大肠癌浸润淋巴细胞Fas配基表达与癌细胞凋亡的关系

目的　研究结肠直肠癌浸润淋巴细胞Fas配基 (FasL)表达与癌细胞凋亡的关系。　方法　用免疫组化方法检测 86例结肠直肠癌及切缘正常大肠组织的Fas蛋白、FasL的表达 ;用TdT酶介导的DNA3′ OH末端 缺口染色方法检测癌细胞的调亡指数 ,并分析浸润淋巴细胞FasL表达与癌细胞凋亡指数的关系。　结果　 86例结肠直肠癌组织呈Fas蛋白染色阳性 5 3例 (6 1 6 % ) ,正常大肠组织则未见染色 ,癌组织旁浸润淋巴细胞FasL染色阳性 6 9例 (80 2 % )。癌组织Fas蛋白阳性组中 ,浸润淋巴细胞FasL染色程度与癌细胞凋亡指数呈正相关 (P <0 0 1) ,FasL染色越强 ,凋亡癌细胞越多。Fas蛋白阴性组中则不存在这种相关性。　结论　癌旁浸润淋巴细胞通过其表面的FasL与大肠癌细胞表面的Fas蛋白作用 ,是诱导癌细胞凋亡的途经之一。提高癌细胞Fas蛋白的表达及应用特异性Fas抗体 ,将为大肠癌治疗提供新思路。     

103钯放射性支架对Fas基因表达影响及与胆管癌细胞凋亡关系研究

目的探讨^103钯（^103Pd）放射性支架释放的γ射线对Fas表达的影响以及与胆管癌细胞凋亡的关系及意义。方法培养瓶中胆管癌细胞消化后形成细胞悬液，采用血细胞计数法计数细胞，按1×10^5／ml的浓度分装到2ml塑料冻存管中。分别设置普通支架组及^103Pd支架组，应用TUNEL法和免疫组化染色方法检测γ射线诱导胆管癌细胞凋亡的情况和Fas表达的情况。结果^103Pd支架组胆管癌细胞的Fas表达较普通支架组明显增高（P〈0．05），且胆管癌细胞出现明显细胞凋亡，凋亡细胞数明显高于普通支架组（P〈0．01）。结论Fas表达水平与胆管癌细胞凋亡的发生有关，^103Pd放射性支架可能通过增强Fas表达，促进胆管癌细胞凋亡，从而为临床应用^103Pd放射性支架治疗胆管癌提供理论依据。     

（103）钯放射性支架诱导人胆管癌细胞凋亡及对Fas基因表达的影响

目的探讨103钯（Pd）放射性支架释放的γ射线对Fas基因表达的影响以及与胆管癌细胞凋亡的关系及意义。方法建立人胆管癌裸鼠移植瘤模型;将建模成功的裸鼠分为实验组和对照组,实验组置入37MBq103Pd胆道放射性支架,对照组置入普通金属支架;支架置入后10d计算肿瘤体积;应用TUNEL法检测胆管癌细胞凋亡的情况;免疫组化染色方法检测胆管癌细胞中Fas基因表达的情况。结果实验组肿瘤体积较对照组增长明显缓慢（P〈0.05）;胆管癌细胞的Fas基因表达阳性率较对照组明显增高（P〈0.05）,且胆管癌细胞出现明显凋亡,细胞凋亡率明显高于对照组（P〈0.01）。结论103Pd放射性支架可诱导人胆管癌裸鼠移植瘤中胆管癌细胞凋亡,明显抑制胆管癌细胞生长,其可能通过增强Fas基因表达促进胆管癌细胞凋亡,可能是103Pd放射性支架治疗胆管癌的重要机理之一,进一步为临床应用103Pd放射性支架治疗胆管癌提供了理论依据。     

联合应用阿霉素和神经节苷脂GD3抑制SMMC-7721肝癌细胞的生长

目的联合应用阿霉素和神经节苷脂GD3能否促进肝癌细胞的凋亡和抑制细胞的生长。方法使用MTT法比较GD3和阿霉素联合处理的与单独使用阿霉素的肝癌细胞生长曲线的差异。通过细胞凋亡ELISA法检测GD3和阿霉素联合处理的肝癌细胞凋亡状况。通过Western blot方法了解GD3和阿霉素联合处理肝癌细胞的胞浆蛋白中caspase 3酶的表达情况,并检测caspase 3的活性。结果与单独使用阿霉素的肝癌细胞比较,低浓度GD3预处理后的肝癌细胞使用阿霉素处理后凋亡显著增加,肝癌细胞的生长受到明显抑制。低浓度的GD3和阿霉素联合应用后导致肝癌细胞中caspase 3酶的表达增加,活性增强。结论神经节苷脂GD3增强阿霉素诱导肝癌细胞凋亡的作用,抑制肝癌细胞的生长,从而增加了阿霉素的抗肝癌敏感性。     

维生素E琥珀酸酯对人肝癌细胞凋亡和侵袭性

目的研究维生素E琥珀酸酯（VES）诱导体外培养人肝癌细胞凋亡以及对肝癌细胞侵袭性的抑制作用。方法自2004年至2005年7月,体外培养SMMC7721人肝癌细胞系,在培养液中加入VES共育,用等量的生理盐水作为对照组。培养6、24、48h后采用流式细胞仪检测肝癌细胞的凋亡,小孔趋化试验检测肝癌细胞的体外侵袭性变化。结果在加入VES后肝癌细胞的凋亡率与对照组相比明显增加。在48h加入VES的实验组与对照组相比,肝癌细胞的小孔趋化试验明显受到抑制。结论VES具有体外诱导人肝癌细胞系凋亡以及抑制肝癌细胞体外侵袭性的作用,是潜在的抗肿瘤因子。     

维生素E琥珀酸酯对人肝癌细胞凋亡和侵袭性的影响研究

目的 研究维生素E琥珀酸酯(VES)诱导体外培养人肝癌细胞凋亡以及对肝癌细胞侵袭性的抑制作用.方法 自2004年至2005年7月,体外培养SMMC7721人肝癌细胞系,在培养液中加入VES共育,用等量的生理盐水作为对照组.培养6、24、48 h后采用流式细胞仪检测肝癌细胞的凋亡,小孔趋化试验检测肝癌细胞的体外侵袭性变化.结果 在加入VES后肝癌细胞的凋亡率与对照组相比明显增加.在48 h加入VES的实验组与对照组相比,肝癌细胞的小孔趋化试验明显受到抑制.结论 VES具有体外诱导人肝癌细胞系凋亡以及抑制肝癌细胞体外侵袭性的作用,是潜在的抗肿瘤因子.     

Fas/FasL在乳头状甲状腺癌组织中的表达

目的：探讨凋亡相关基因Fas/FasL在甲状腺癌组织中的表达与细胞凋亡的关系。方法：采用流式细胞技术检测43例乳头状甲状腺癌癌细胞凋亡及相关基因Fas及FasL表达。同时检测Fas及FasL在甲状腺癌组织肿瘤浸润淋巴细胞（TIL）中的表达。结果：43例乳头状甲状腺癌细胞中Fas低表达者19例,细胞凋亡率为3．71％,高表达者24例,细胞凋亡率为7．26％,高表达组癌细胞凋亡率明显高于低表达组（P＜0．05）,癌细胞中FasL表达明显高于甲状腺腺瘤组织（P＜0．01）,乳头状甲状腺癌组织TLL表达Fas及FasL的水平明显高于甲状腺腺瘤组织（P＜0．01）。结论：Fas系统参与乳头状甲状腺癌细胞中细胞凋亡的调节,Fas/FasL表达异常使甲状腺癌癌细胞逃避免疫监视,诱导Fas敏感的TIL凋亡,有助于癌细胞发生浸润及转移。     

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目的 探讨一氧化氮（nitric oxide,NO)在胆管癌细胞凋亡中的作用机制。方法 通过细胞培养,采用流式细胞仪等测定体外培养胆管癌细胞QB内NO含量,同时检测不同水平NO细胞内细胞凋亡率。结果 低浓度NO对胆管癌QB细胞凋亡无影响,高浓度的NO明显促进QB细胞的凋亡发生。结论 NO对胆管癌细胞内细胞凋亡具有双重作用。     

距骨缺血坏死手术治疗进展

距骨缺血坏死是距骨骨折的一种严重并发症,近年来发病率呈上升趋势。虽然大多数表现较典型,但如对其认识不足,术式选择不当,较易误诊、误治。本文从距骨生理解剖,距骨坏死的临床表现以及各种手术方法等方面做一简单介绍。     

Measurement of varus-valgus and internal-external rotational knee laxities in vivo--Part II: relationship with anterior-posterior and general joint laxity in males and females.

We examined sex differences in general joint laxity (GJL), and anterior-posterior displacement (ANT-POST), varus-valgus rotation (VR-VL), and internal-external rotation (INT-EXT) knee laxities, and determined whether greater ANT and GJL predicted greater VR-VL and INT-EXT. Twenty subjects were measured for GJL, and scored on a scale of 0-9. ANT and POST were measured using a standard knee arthrometer at 133 N. VR-VL and INT-EXT were measured using a custom joint laxity testing device, defined as the angular displacements (deg) of the tibia relative to the femur produced by 0-10 Nm of varus-valgus torques, and 0-5 Nm of internal-external torques, respectively. INT-EXT were measured during both non-weight-bearing (NWB) and weight-bearing (WB = 40% body weight) conditions while VR-VL were measured NWB. All laxity measures were greater for females compared to males except for POST. ANT and GJL positively predicted 62.5% of the variance in VR-VL and 41.8% of the variance in WB INT-EXT. ANT was the sole predictor of INT-EXT in NWB, explaining 42.3% of the variance. These findings suggest that subjects who score higher on clinical measures of GJL and ANT are also likely to have greater VR-VL and INT-EXT knee laxities.     

Enhancement of Apo2L/TRAIL (tumor necrosis factor-related apoptosis-inducing ligand)-induced apoptosis in non-small cell lung cancer cell lines by chemotherapeutic agents without correlation to the expression level of cellular protease caspase-8 inhibitory protein.

OBJECTIVE: Apo2L/tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potential anticancer drug that promotes apoptosis specifically in tumor cells. Because not all cancer cells are susceptible to Apo2L/TRAIL, the aim of our study was to determine whether non-small cell lung cancer cells can be sensitized by chemotherapeutic agents for Apo2L/TRAIL-induced apoptosis. In addition, endogenous expression levels of the caspase-inhibiting cellular protease caspase-8 inhibitory protein (C-FLIP) were measured to investigate partial resistance to Apo2L/TRAIL. METHODS: Six human lung cancer cell lines (A549, NCI-H358, Calu1, Calu6, SkMes1, and SkLu1) were incubated with soluble Apo2L/TRAIL and two different concentrations each of cisplatin, paclitaxel, doxorubicin, 5-fluorouracil, and camptothecin. After 24 hours the rate of apoptosis was measured by annexin V/propidium iodide staining followed by FACScan analysis. Expression levels of C-FLIP in cell lines and lung cancer biopsy specimens were determined by Western blotting. RESULTS: Treatment of lung cancer cells with Apo2L/TRAIL alone resulted in apoptotic cell death in four cell lines (P <.001). Combining Apo2L/TRAIL and chemotherapeutic agents enhanced the rate of apoptosis significantly. Statistical analysis revealed a synergistic effect of Apo2L/TRAIL in combination with 1.8 mmol/L camptothecin and 100 micromol/L cisplatin, each in four of the six cell lines (P <.002). Western blot analysis showed that sensitization to Apo2L/TRAIL did not correlate with the expression of cellular protease caspase-8 inhibitory protein. Furthermore, no increased cellular protease caspase-8 inhibitory protein levels relative to those in normal lung tissue could be found in non-small cell lung cancer specimens from 12 patients. CONCLUSION: Apo2L/TRAIL-induced apoptosis in non-small cell lung cancer cell lines is significantly enhanced by chemotherapeutic agents. Resistance and sensitization to Apo2L/TRAIL are not correlated with the endogenous expression level of cellular protease caspase-8 inhibitory protein, implying that in non-small cell lung cancer other mechanisms are responsible for inhibition of the Apo2L/TRAIL pathway. Even though the molecular mechanism remains unclear, the combination of Apo2L/TRAIL with chemotherapy may be a promising treatment modality for non-small cell lung cancer.     

stTRAIL—MSC靶向促进肝癌大鼠RFA过渡区残癌细胞凋亡的体内研究

目的：探讨stTRAIL-MSC靶向促进肝癌大鼠RFA过渡区残癌细胞凋亡的作用。方法：对150只雄性SD大鼠,采用N1S1肿瘤细胞建立肝荷瘤大鼠模型,并随机将其分为stSTRAL-MSC组、MSC组、Vector-MSC组、培养液组以及空白对照组,每组30只,在接受治疗前1周给以注射相应的制剂,在接受RFA治疗后12h、24h、48h以及1周时每组取6只处死并进行相关检查（空白组除外）,同时比较空白组与RFA组其生存时间。结果：stTRAIL-MSC在接受RFA治疗后其肿瘤体积变化、过渡区肿瘤细胞凋亡指数、大鼠肿瘤毁损体积、过渡区肿瘤细胞（ki-67染色）增殖指数等比较明显优于其他几组,而且在对caspase-3、caspase-8、Bcl-2水平检测时发现,stTRAIL-MSC其caspase-、3caspase-8水平明显高于其他四组,而Bcl-2水平低于其他四组,对接受RFA以及空白对照组大鼠的生存情况分析发现,采用RFA治疗的大鼠其生存时间明显高于空白对照组。结论：stTRAIL-MSC靶向通过提高cas-pase蛋白酶级联反应继而达到促进肝癌大鼠RFA过渡区残癌细胞凋亡。     

STAT1 Activation-Induced Apoptosis of Esophageal Squamous Cell Carcinoma Cells In Vivo

Background The induction of apoptosis might be a promising treatment for cancers refractory to conventional therapies, such as esophageal cancer. In this study, we examined whether epidermal growth factor–induced growth inhibition results from apoptosis of esophageal squamous cell carcinoma (SCC) cells as a result of STAT1 activation and evaluated whether interferon gamma (IFN-γ) can induce apoptosis of cancer cells in vivo. Methods To assess the function of STAT1, we established stable transfectants expressing dominant-negative STAT1. Apoptosis was assessed by several experimental techniques, including flow cytometry. Differentiation was evaluated by Western blot test with involucrin used as a marker. In vivo, cancer cells were injected into male BALB/c nu/nu mice. Two weeks later, the mice started to receive injections of IFN-γ or saline into a tail vein four times per week. Concentrations of IFN-γ in the tumors were analyzed by enzyme-linked immunosorbent assay. Apoptosis was evaluated by TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling) staining. Results Epidermal growth factor inhibited the growth of esophageal SCC cells by causing apoptosis through several pathways involving STAT1 activation. IFN-γ induced the apoptosis of cancer cells, but it also promoted the differentiation (not apoptosis) of primary cultured cells derived from normal esophageal epithelium. IFN-γ also inhibited the growth of xenograft tumors of esophageal SCC cells in vivo. Conclusions Our results suggest that IFN-γ is one candidate for cytokine-based therapy of cancer. IFN-γ–induced STAT1 activation might be involved in the apoptosis of esophageal SCC cells and in the terminal differentiation of normal squamous cells. Further studies of STAT1 signaling pathways may provide the basis for new targeted therapeutic strategies for esophageal SCC.     

Content of adenine nucleotide translocator mRNA in insulin-producing cells of different functional states

This study was undertaken to characterize the expression of the gene coding for the adenine nucleotide translocator (ANT) in the insulin-producing beta-cell and to study any possible relationship between its expression and the functional state of the beta-cell. Adult and fetal rat pancreatic islets were prepared and cultured under different conditions in vitro. The total RNA from these islets and from the insulin-producing RINm5F cells was isolated and analyzed by the Northern blot technique via a cDNA clone (pAAC-9) coding for the bovine ANT. We found that a 1600-base pair (bp) mRNA hybridizing to the pAAC-9 clone could be detected in RINm5F cells, and a 1450-bp mRNA was similarly observed in the islets. These sizes correspond well to previously reported forms of mRNA for the ANT observed in other tissues. When comparing the intensities of the pAAC-9 hybridizing bands of the different islet groups, it was observed that fetal islets contained less of this mRNA than adult islets. Furthermore, the content of the ANT mRNA in adult islets cultured at a high glucose concentration was increased compared with islets cultured at a low glucose concentration. Finally, streptozocin-treated islets, which display an impaired glucose-sensitive insulin release after 6 days in culture, also contained less of this mRNA than the control islets. We conclude that pancreatic islet cells express an mRNA that appears to be highly homologous to the bovine ANT and that the contents of this mRNA increases with the functional status of the beta-cell. It is furthermore suggested that defects in the expression of this gene may be associated with impaired glucose sensitivity.     

Fas and Fas-ligand expression in human pancreatic cancer

OBJECTIVE: To investigate Fas and FasL expression in pancreatic tissues and cultured pancreatic cancer cell lines, and to assess the ability of anti-Fas antibodies to induce apoptosis. SUMMARY BACKGROUND DATA: Activation of the Fas receptor by Fas-ligand (FasL) results in apoptosis, and dysregulation of this pathway may contribute to abnormal cell proliferation. METHODS: Northern blotting and immunohistochemistry were used to compare Fas and FasL expression in normal and cancerous tissues. DNA 3'-OH end labeling was used to detect apoptotic cells. The effects of Fas activation on cell growth and signaling pathways were investigated in culture. RESULTS: Pancreatic cancers exhibited increased Fas RNA levels, whereas FasL mRNA levels were similar in both groups. Despite the colocalization of Fas and FasL in the cancer cells, an apoptotic signal was present in approximately 10% of these cells in only 2 of 16 cancer samples. Fas and FasL were coexpressed in all four cell lines, whereas Fas-associated phosphatase 1 was below the level of detection in all cell lines. Only COLO-357 cells underwent apoptosis after Fas activation. Apoptosis was associated with enhanced activation of jun kinase (JNK) and p38 mitogen-activated protein kinase (MAPK). In the presence of actinomycin D, Fas antibody also induced apoptosis in the other three cell lines. CONCLUSIONS: These results suggest that pancreatic cancer cells are resistant to Fas-mediated apoptosis by mechanisms excluding receptor downregulation or Fas-associated phosphatase upregulation and raise the possibility that Fas-mediated apoptosis may be dependent on the activation of the JNK/p38 MAPK pathway in these cells.