Tamoxifen (TMX)/Fas induced growth inhibition of human cholangiocarcinoma (HCC) by gamma interferon (IFN-gamma)

OBJECTIVES: To evaluate the response of human cholangoicarcinoma cells to TMX treatment through the Fas pathway by pretreatment with IFN-gamma. SUMMARY BACKGROUND DATA: Cholangiocarcinoma remains one of the most difficult tumors to treat in clinical medicine. Currently, there are no effective chemotherapy treatments for this disease. Surgery offers the only opportunity for a cure, with the majority of patients failing to qualify for such treatment. This study seeks to evaluate a potential new modality for treatment of this disease. METHODS: Human cholangiocarcinoma cells were treated with anti Fas mab and sorted to two populations (Fas-positive and Fas-negative) by FAC analysis. In vitro individual cell populations were pretreated with IFN-gamma 250 units/mL x 18hs. The treated cells assayed for caspase 3, 7, 8, Bak, and for apoptosis with Annexin V after treatment with or without TMX. In Vivo 2 x 106 5 SK-ChA-1 Fas-negative cells were injected into nude mice for development of tumor xenografts. Mice received either no treatment or intra tumor IFN-gamma and/or intra peritoneal TMX. RESULTS: More than 90% (90% +/- 3.5%) of Fas-positive and 70% (71 +/- 2.3%) of Fas-negative cells underwent apoptosis after TMX treatment when pretreated with IFN-gamma. In contrast, TMX alone and IFN-gamma alone stimulated apoptosis by only 22% (22 +/- 3%) P <.00013, and 17% (17 +/- 2%) P <.0001 in Fas-ve cells respectively. In vivo human cholangiocarcinomas xenograft growth was significantly inhibited by a combination of TMX + IFN-gamma compared to controls P <.0007. CONCLUSION: TMX exposure to human cholangiocarcinoma after pretreatment with IFN-gamma allows for induction of apoptosis in vitro and significant inhibition tumor xenograft growth. The combination of these two compounds may provide novel treatment regimen for cholangiocarcinoma.     

（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。     

阿霉素上调肿瘤细胞Fas基因表达并诱导凋亡的实验研究

目的 研究化疗药物阿霉素(ADM)对肿瘤细胞Fas凋亡基因的调控，并探讨其致凋亡机制。方法 逆转录一聚合酶链反应(RT-PCR)比较经。ADM处理前后肿瘤细胞胃癌ADM细胞系SGG-7901、MGC-803和肺癌细胞系A-549的Fas mRNA的表达水平，同时观察其对抗Fas抗体的敏感性，并用流式细胞术检测肿瘤细胞凋亡率。结果 所有3种肿瘤细胞在ADM作用下Fas表达水平显著增强，差异有统计学意义(P＜0．05)，同时肿瘤细胞凋亡率明显增加(凋亡率分别为40．3％、43．6％、54．4％)。ADM能增加肿瘤细胞对Fas抗体的敏感性。结论 ADM上调肿瘤细胞Fas的表达，ADM与Fas抗体联合应用将增强肿瘤细胞的凋亡率，有一定的抗肿瘤临床应用前景。     

Cisplatin augments cytotoxic T-lymphocyte-mediated antitumor immunity in poorly immunogenic murine lung cancer

OBJECTIVE: Many tumors are poorly immunogenic and resistant to cytotoxic T-lymphocyte-mediated cell lysis. Because cisplatin has been demonstrated to increase tumor cell Fas receptor expression, we hypothesized that cisplatin will enhance cytotoxic T-lymphocyte tumor cell killing and augment the antitumor effect of an active immunotherapy strategy in a poorly immunogenic murine lung cancer model. METHODS: Lewis lung carcinoma cells were exposed to cisplatin in vitro, and Fas receptor expression and apoptosis in response to an agonistic anti-Fas antibody were quantified using flow cytometry. Wild-type and Fas ligand-deficient mice bearing Lewis lung carcinoma flank tumors were then treated with intraperitoneal cisplatin as well as an intratumoral injection of an adenovirus gene transfer vector encoding CD40 ligand. End points included tumor size, animal survival, and Fas expression (determined using immunofluorescence). Cytotoxicity assays were performed using splenocytes from adenovirus gene transfer vector encoding CD40 ligand-treated animals as effectors and cisplatin-treated Lewis lung carcinoma cells as targets. RESULTS: Cisplatin induced heightened expression of Fas receptor on Lewis lung carcinoma cells in vitro and in vivo and enhanced apoptosis in cells exposed to an agonistic anti-Fas antibody. In vivo, the combination of 1 dose of intraperitoneal cisplatin and intratumoral adenovirus gene transfer vector encoding CD40 ligand inhibited tumor growth and prolonged survival compared with adenovirus gene transfer vector encoding CD40 ligand alone, resulting in a higher cure rate. This effect was lost in Fas ligand-deficient mice. Splenocytes from adenovirus gene transfer vector encoding CD40 ligand-treated wild-type mice lysed cisplatin-treated Lewis lung carcinoma cells more efficiently than untreated Lewis lung carcinoma cells, an effect lost in splenocytes from Fas ligand-deficient mice. CONCLUSION: Cisplatin augments the antitumor effect of a cytotoxic T-lymphocyte-mediated immunotherapy strategy, resulting in a higher cure rate than seen with immunotherapy alone. This effect is associated with the enhanced ability of cytotoxic T lymphocytes to lyse tumor cells that have been exposed to cisplatin through Fas/Fas ligand interactions.     

维生素E琥珀酸酯对乳腺癌荷瘤裸鼠的抑瘤作用

目的 检测维生素E琥珀酸酯（VES）对裸鼠荷乳腺癌MCF-7细胞的抑制作用。方法 裸鼠皮下接种MCF-7乳腺癌细胞建立荷瘤裸鼠模型。以150mg／kg（体重）剂量的VES治疗模型鼠共5周。处死动物后测量肿瘤大小。以流式细胞仪检测细胞周期变化和细胞表面Fas和FasL的表达；以免疫组化法检测肿瘤组织Fas和FasL的表达，并以TUNEL法检测细胞凋亡指数。结果 VES对荷乳腺癌裸鼠具有显著的增殖抑制作用。细胞周期分析显示，VES冶疗后肿瘤细胞表现为G0／G1期阻滞，肿瘤组织和细胞表面的Fas和FasL均出现上调，同时细胞的凋亡指数升高。结论 VES对于MCF-7乳腺癌荷瘤裸鼠具有强效的抑制作用，其机制与上调肿瘤细胞Fas／FasL表达，促进细胞凋亡有关。     

Omega-6 fatty acids can inhibit Fas-mediated apoptosis in a human colorectal carcinoma cell line: a potential mechanism for escape from immune surveillance

The Fas/Fas ligand pathway may play an important role in the pathogenesis of colorectal carcinoma by allowing tumor cells to evade host immune defenses. Since dietary fats, in particular omega-6 fatty acids, facilitate tumor development their influence on the Fas/Fas ligand pathway needs to be elucidated. The purpose of this study was to determine the effect of membrane free fatty acid (FFA) alterations on Fas expression and sensitivity. Two human colorectal carcinoma cell lines (CX-1 and CCL-188) were grown in cell culture media supplemented with omega-3 (docosahexanoic acid) and omega-6 (linoleic acid) fatty acids. Membrane alterations were confirmed by gas chromatography (GC). Cell surface Fas expression was determined with flow cytometry using an anti-Fas monoclonal antibody. Sensitivity to Fas mediated apoptosis was measured by now cytometric measurement of fragmented DNA stained with propidium iodide. Appropriate changes in the membrane FFA composition were found by GC. Cell surface Fas expression was unaffected in either cell line. Omega-3 fatty acids did not alter Fas sensitivity for either cell line compared to control (CX-1: 59.7%, +/- 5.4 vs 51.4% +/- 7.1 for control, CCL-188: 54.3% +/- 8.6 vs 51.2% +/- 4.8 for control). Omega-6 fatty acids produced a significant decrease in Fas-mediated apoptosis (CX-1: 34.2% +/- 4.8 and CCL-188: 22% +/- 6.0, p < 0.05 vs control). These data indicate that although membrane FFA alterations did not affect Fas expression, omega-6 fatty acids significantly decreased Fas-mediated apoptosis. This inhibitory effect may protect colorectal carcinoma cells from lymphocyte Fas-mediated cell death.     

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.     

Enhanced cell death in NR-S1 tumor by photodynamic therapy: possible involvement of Fas and Fas ligand system

BACKGROUND AND OBJECTIVES: To investigate the effect of photodynamic therapy (PDT) on cell death in malignant tumor tissue, the frequency of terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL)-positive cells and the possible involvement of Fas and Fas ligand system were evaluated. STUDY DESIGN/MATERIALS AND METHODS: NR-S1 tumor-bearing C3H/HeNCrj mice were treated by PDT with Photofrin(R) (12 mg/kg body weight) and Nd:YAG dye laser (630 nm, 10 Hz, 150 J/cm(2)). Paraffin-embedded tissue sections from the excised tumor tissues at 6, 12, 24, 48 hours after PDT were analyzed by TUNEL for the occurrence of apoptosis and by immunohistochemistry for Fas and Fas ligand (FasL) expression. TUNEL-positive cells as well as Fas- or FasL-positive cells were counted and expressed as a percentage of positive cells per total cells. RESULTS: Based on the percent area of tumor necrotic foci, the most effective conditions for PDT were first determined. Under these conditions, PDT increased the number of TUNEL-positive tumor cells at 12 hours after irradiation. In parallel with the increase in TUNEL-positive cells, Fas-positive tumor cells were also found in the same area where many TUNEL-positive tumor cells were found. The expression of Fas ligand was found in the tumor cells surrounding TUNEL-positive cells on serial sections. A significant increase in FasL-positive lymphocytes was observed at 12 hours, whereas the infiltration of such lymphocytes into the area where TUNEL-positive tumor cells were observed was rare. CONCLUSION: The possible role of Fas/FasL system in the cell death induced by PDT with Photofrin(R) and Nd:YAG dye laser was suggested. Moreover, the role of infiltrated lymphocytes seemed not to be so much in this model.     

腹主动脉瘤患者T淋巴细胞Fas系统功能的研究

目的 通过分析Fas诱导的外周血活化T淋巴细胞的凋亡来评价腹主动脉瘤(abdominal aortic aneurysm,AAA)患者Fas系统的功能. 方法采用细胞培养方法,应用Fas单克隆抗体激活Fas通路,分析20例AAA患者和15例腹主动脉硬化闭塞症(aortic atherosclerotic occlusive disease,AOD)患者以及25例正常人外周血T淋巴细胞Fas系统的功能,并计算细胞的存活率和死亡率. 结果 AOD组和正常人组中Fas诱导的活化T细胞发生很强的凋亡反应,而AAA患者组则不明显.AAA组细胞存活率(98.9%±10.3%)明显高于AOD组(58.9%±15.2%)和正常人组(59.4%±12.9%,P＜0.01).该结果与采用FICT-Annexin-V和propidium iodide染色及流式细胞仪检测分析细胞凋亡的结果一致.T细胞Fas功能的缺陷并不是Fas表达减少所致,而且这种功能缺陷具有Fas特异性,因为采用甲基强的松龙均可诱导细胞凋亡.结论 AAA患者T淋巴细胞中存在Fas系统功能缺陷,这种缺陷可能是主动脉瘤发展的重要影响因素.     

Impaired Fas-induced apoptosis of T lymphocytes in patients with abdominal aortic aneurysms

OBJECTIVE: Homeostasis of the immune system is maintained by apoptotic elimination of potentially pathogenic autoreactive lymphocytes. Emerging evidence shows that Fas-mediated apoptosis is impaired in activated lymphocytes from patients with autoimmune disease. The aim of this work was to assess apoptosis mediated by the cell death receptor Fas in peripheral T lymphocytes from patients with abdominal aortic aneurysms (AAA). METHODS: The apoptotic pathway was triggered by anti-Fas monoclonal antibodies in cultured and activated peripheral T-cell lines from 20 AAA patients with control groups of 15 patients with aortic atherosclerotic occlusive disease (AOD) and 25 healthy individuals. Cell survival and death (apoptosis) rate were assessed. RESULTS: Cross-linkage of Fas receptor exerted a strong apoptotic response on T cells from AOD patients and healthy controls, but a much less pronounced effect on T cells from AAA patients. The evaluation of cell survival rate showed a significantly higher percentage in AAA group (98.9% +/- 10.3%) than in the AOD subjects (58.9% +/- 15.2%) or the healthy group (59.4% +/- 12.9%; P < .001). Apoptosis assessment by annexin V and propidium iodide staining and flow cytometry showed similar results. The defect in AAA group was not due to decreased Fas expression, since Fas was expressed at normal levels. Moreover, it specifically involved the Fas system because cell death was induced in the normal way by methylprednisolone. Complementary DNA sequencing identified no causal Fas gene mutation, but two silent single nucleotide polymorphisms with higher frequency were found in the AAA group. CONCLUSIONS: Fas-induced apoptosis in activated T cells from AAA patients is impaired. This may disturb the normal down-regulation of the immune response and thus provide a new insight into possible mechanisms and routes in the pathogenesis of AAA.     

Critical roles for JNK, c-Jun, and Fas/FasL-Signaling in vitamin E analog-induced apoptosis in human prostate cancer cells

BACKGROUND: Alpha-tocopherol ether-linked acetic acid (alpha-TEA), an analog of vitamin E (RRR-alpha-tocopherol), is a potent pro-apoptotic agent for human cancer cells in vivo and in vitro. METHODS: alpha-TEA-induced apoptosis was investigated in LNCaP and PC-3 human prostate cancer cells. Apoptosis was measured by DAPI-staining and FACS analyses of the sub-G1 fraction. Signaling molecules involved in apoptosis were measured by Western immunoblot analyses with or without prior immunoprecipitation, FACS analyses of cell surface membrane expression, RT-PCR analyses of mRNA levels, and chromatin immunoprecipitation. Functional significance was determined using siRNAs, dominant negative mutant, chemical inhibitor, or neutralizing antibody. RESULTS: Alpha-TEA treatment increased Fas and Fas ligand mRNA and protein levels; as well as, levels of cell surface membrane Fas in both cell lines. Blockage of Fas signaling attenuated alpha-TEA-induced apoptosis. alpha-TEA treatment also produced prolonged, elevated levels of activated (phosphorylated) c-Jun N-terminal kinase (JNK) and its substrate c-Jun, both of which were demonstrated to be necessary for alpha-TEA-induced apoptosis. Chromatin immunoprecipitation results showed binding of c-Jun to the promoters of both Fas and FasL in alpha-TEA treated cells. Investigations of alpha-TEA-triggered apoptosis showed dual signaling from Fas with essential roles for both FADD and Daxx with FADD initiating the classical pathway mediated by caspase-8 activation and Daxx initiating an alternate pathway involving activation of JNK, c-Jun, and increased levels of Fas and FasL. CONCLUSIONS: Collectively, data support critical roles for JNK, c-Jun, and dual signaling from Fas/FasL via FADD and Daxx in alpha-TEA-induced apoptosis of human prostate cancer cells.     

CD4+ cells play a major role in xenogeneic human anti-pig cytotoxicity through the Fas/Fas ligand lytic pathway

BACKGROUND: In this study, the role of cell-mediated cytotoxicity by human leukocytes against pig endothelial cells was examined in vitro. The aim was to determine which cell subsets were responsible for this phenomenon and which pathways were involved in cell lysis. METHODS: Primed human peripheral blood mononuclear cells (PBMC) or purified CD4+ or CD8+ T cells were used in a cell-mediated cytotoxicity assay in which cytotoxicity of an SV40 transformed porcine endothelial cell (EC) line (SVAP) was determined by Annexin V binding. RESULTS: Human PBMC demonstrated specific lysis of porcine EC that was proportional to the effector: target ratio. CD4+ T cells accounted for >60% of this lysis, whereas CD8+ T cells accounted for <20%. CD4+ T cell-mediated lysis depended on direct recognition of porcine major histocompatibility complex class II molecules as inhibition of swine leukocyte antigen class II on porcine EC-inhibited CD4+ T cell cytotoxicity. This lysis was mediated through the Fas/FasL pathway as addition of anti-Fas and/or anti-FasL antibody profoundly inhibited antiporcine lysis. In addition, FasL gene expression was detected in primed PBMC and CD4+ T cells by RT-PCR, whereas granzyme B gene expression was not. Primed CD4+ T cells demonstrated high level FasL protein by Western blotting and two-color FACS analysis, whereas NK cells and CD8+ T cells did not. Finally, recombinant human FasL induced apoptosis in Fas expressing porcine EC cells, demonstrating that human FasL interacted with and activated Fas on porcine EC cells. CONCLUSIONS: In conclusion, human to pig cell-mediated cytotoxicity was mediated predominantly by CD4+ T cells through the Fas/FasL pathway of apoptosis. These results suggest that direct cytotoxicity by xenoreactive CD4+ T cells may be one of several effector mechanisms involved in cellular xenograft rejection.     

抗Fas单克隆抗体诱导人胃癌细胞系SGC-7901细胞凋亡的研究

目的 探讨抗Fas单克隆抗体诱导胃癌细胞凋亡的规律及在胃癌治疗中的意义。方法 应用细胞形态观察、琼脂糖凝胶电泳、流式细胞光度术检测抗Fas单克隆抗体对胃癌细胞SGC-7901增殖周期的影响以及对细胞杀伤作用的方式，并检测了SGC-7901细胞表面bcb2的表达情况。结果 抗Fas单克隆抗体有阻滞细胞周期、通过诱发凋亡而抑制肿瘤细胞生长的作用。经抗Fas单克隆抗体处理后，SGC-7901细胞表面bcl-2蛋白表达无明显变化。结论 抗Fas单克隆抗体可以诱导胃癌细胞系SGC-7901细胞凋亡，抗Fas单克隆抗体诱导胃癌细胞凋亡与bcl-2表达无关。     

Differential involvement of the Fas receptor/ligand system in p53-dependent apoptosis in human prostate cancer cells

BACKGROUND: The objective of this study was to characterize the involvement of the Fas receptor/ligand system in p53-dependent apoptosis in human prostate cancer cells. METHODS: The effects of adenovirus-mediated p53 gene transfer (Ad5CMV-p53) into human prostate cancer LNCaP, DU145, and PC3 cells on their growth, apoptosis and Fas receptor/ligand expression were examined by the MTT assay, DNA fragmentation assay, and Northern blot analysis, respectively. The sensitivity of these cells to an agonistic anti-Fas receptor antibody (CH11) and the effects of an antagonistic anti-Fas ligand antibody (4H9) on Ad5CMV-p53-induced apoptosis were analyzed by the MTT assay and DNA fragmentation assay. RESULTS: Ad5CMV-p53 treatment resulted in substantial growth inhibition, induction of apoptosis and up-regulation of Fas receptor as well as Fas ligand mRNA expression in LNCaP, DU145 and PC3 cells. Despite the abundant expression of Fas receptor in all of these cells, CH11 induced apoptosis only in PC3 cells. Furthermore, 4H9 partially blocked the apoptosis induced by Ad5CMV-p53 in PC3 cells, but not in LNCaP and DU145 cells. CONCLUSIONS: The Fas receptor/ligand system is differentially involved in p53-dependent apoptosis in prostate cancer cells; therefore, reintroduction of wild-type p53 into prostate cancer cells may induce apoptosis through Fas receptor/ligand interaction as well as through an alternative pathway.     

Expression of Fas receptor on disc cells in herniated lumbar disc tissue

STUDY DESIGN: The expression of Fas receptor, an apoptosis-related protein, on disc cells was examined in surgically obtained disc specimens. OBJECTIVE: To assess the fate of disc cells in herniated disc tissue and the difference in the degree of expression of the Fas receptor between contained and noncontained discs. SUMMARY OF BACKGROUND DATA: Little is known about the fate of disc cells after herniation. METHODS: Twenty-three herniated lumbar disc specimens were classified into contained discs (protrusion or subligamentous extrusion; n = 9) and noncontained discs (transligamentous extrusion or sequestration; n = 14). All specimens were stained using the avidin-biotin-peroxidase complex method. The percentage of disc cells positive for Fas receptor was calculated and compared with clinical and radiologic data. RESULTS: There was a significant difference in the percentage of Fas-positive disc cells between the contained and noncontained discs (8.44 vs.- 14.29;P = 0.044). The percentage of Fas-positive disc cells correlated significantly with the patient's age (r = 0.455, P = 0.029), but not with the degree of disc degeneration on magnetic resonance imaging (r = 0.252, P = 0.214). CONCLUSION: This is the first study to identify the expression of Fas receptor on disc cells in herniated disc tissue. The results show that the disc cells after herniation may undergo Fas-mediated apoptosis and that the degree of expression of Fas receptor differs depending on the type of herniation.     

MAPK13调控胆管癌细胞增殖、侵袭和上皮间质转化进程的机制研究

目的 本研究旨在分析丝裂原活化蛋白激酶13(MAPK13)在胆管癌组织及细胞中的表达水平并探讨其在胆管癌细胞增殖、侵袭和上皮间质转化(EMT)中的作用。方法 通过微阵列分析筛选胆管癌组织中差异表达的mRNA,通过实时荧光定量PCR(qRT-PCR)、蛋白印迹及免疫组化染色验证MAPK13在肿瘤组织和细胞中的表达情况。进一步通过细胞转染构建MAPK13敲低(si-MAPK13)后,通过细胞活力实验检测胆管癌细胞增殖能力,流式细胞仪检测细胞凋亡,Transwell法检测细胞的迁移和侵袭性。最后,通过裸鼠成瘤实验检测MAPK13对体内成瘤及相关蛋白表达的影响。结果 本研究通过微阵列分析发现MAPK13在肿瘤组织中的表达存在显著差异,进一步通过qRT-PCR、蛋白印迹和免疫组化染色分析发现,与癌旁组织比较,MAPK13在mRNA和蛋白质水平的表达情况在肿瘤组织及细胞中均显著升高且差异具有统计学意义(t=27.92,6.90;均P<0.01)。进一步细胞活力检测表明,抑制MAPK13的表达后细胞增殖数量明显减少(F=32.13,P<0.01)。Transwell实验则显示细胞迁移和侵...     

胞嘧啶脱氨酶基因、单纯疱疹病毒胸苷激酶基因治疗裸鼠肝门部胆管癌皮下移植瘤的研究

目的　观察胞嘧啶脱氨酶基因 (CD)、单纯疱疹病毒胸苷激酶基因 (HSV tk)共表达对裸鼠人肝门部胆管癌皮下移植瘤的治疗作用。方法　用转染有双自杀基因的包装细胞PA3 17产生的病毒上清 ,感染裸鼠肝门部胆管癌皮下移植瘤 ,给予前体药物 5 氟胞嘧啶 (5 FC) 5 0 0mg/kg体重和 /或无环鸟苷 (GCV ) 10 0mg/kg体重后 ,1次 /d ,共 10d ,观察肿瘤的生长情况。 结果　双自杀基因在裸鼠肝门部胆管癌皮下移植瘤中稳定表达 ,转基因组肿瘤的生长明显受到抑制 (P <0 .0 1) ;CD tK / 5 FC GCV组、CD/ 5 FC组、tK/GCV组抑瘤率分别为 88.3 1%、63 .5 2 %、65 .0 5 %。双自杀基因组与单自杀基因比较差异有显著性 (P <0 .0 5 )。结论　双自杀基因可明显抑制裸鼠肝门部胆管癌皮下移植瘤生长 ,较单一自杀基因有更强的抗肿瘤作用