重组人血管内皮抑素联合重组人P53腺病毒对裸鼠乳腺癌移植瘤的抑制

摘 要 目的: 探讨重组人血管内皮抑素（recombinant human endostatin,ES）联合重组人P53腺病毒（rAd/P53）对乳腺癌细胞MCF7裸鼠移植瘤的抑制作用。方法：建立裸鼠荷人乳腺癌模型，随机分为对照组、ES组、rAd/P53组和两药联合组,分别给予相应治疗；观察肿瘤生长，免疫组化法检测肿瘤组织微血管密度（microvessel density，MVD）和血管内皮生长因子（vascular endothelial growth factor, VEGF）的表达。结果：各治疗组均可明显抑制移植瘤生长， ES组、rAd/P53组和联合用药组的抑瘤率分别为51.67％、48.74％和75.54％，联合用药组抑瘤作用最为显著（P<0.05）。对照组、ES组、rAd/P53组与联合用药组的移植瘤组织MVD分别为31.17±2.48、20.33±4.84、22.33±3.88及12.50±2.74，VEGF 表达H评分分别为45.33±589、35.83±546、33.67±4.80及22.33±4.41，联合用药组的MVD和VEGF表达显著低于其余各组（P<0.01）。 结论：重组人ES联合rAd/P53治疗可显著抑制裸鼠乳腺癌移植瘤的生长及微血管生成。     

重组人血管内皮抑素联合紫杉醇对裸鼠MKN-45胃癌抗肿瘤效应的实验研究

目的探讨重组人血管内皮抑素联合紫杉醇对裸鼠原位移植人胃癌抗肿瘤效应及作用机制,为进一步的临床研究提供理论依据。方法采用人胃癌细胞株MKN-45完整组织块作裸鼠原位移植,建立裸鼠胃癌模型,种植后从体表可以触及肿瘤,将32只人胃癌原位种植的裸鼠随机分为重组人血管内皮抑素组(重组人血管内皮抑素15 mg/kg),单一化疗组(选用紫杉醇20 mg/kg),联合用药组(重组人血管内皮抑素15 mg/kg+紫杉醇20 mg/kg)及对照组(等体积生理盐水)4组。给药后取肿瘤组织,测量原位肿瘤重量,计算抑瘤率、肿瘤细胞凋亡、肿瘤微血管密度(MVD)以及血清血管内皮细胞生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)的表达,观察肿瘤细胞腹膜、肝、其他脏器转移及腹水情况。结果联合用药组瘤重、血清VEGF和bFGF、MVD等较单一化疗组及对照组明显降低(P0.01或P0.05),凋亡指数较单一化疗组明显增高(P0.01或P0.05)。结论重组人血管内皮抑素联合紫杉醇能明显抑制裸鼠原位移植人胃癌的生长和转移,显示出较好的协同抗肿瘤作用。     

重组人p53腺病毒注射液(rAd/p53)治疗小鼠乳腺癌的实验研究

目的 探讨重组人p53腺病毒注射液(rAd/p53)不同用药方式对荷瘤小鼠乳腺癌的治疗效果.方法 细胞毒性实验,流式细胞术观察rAd/p53对乳腺癌细胞的体外抑制作用及促凋亡作用.建立乳腺癌小鼠模型,不同方式瘤内注射rAd/p53,观察对肿瘤的生长情况的影响,并对肿瘤组织进行免疫组化检查微血管密度(MVD).结果 rAd/p53体内外均可显著抑制乳腺癌细胞Ca761的生长,且早期连续瘤内给药效果好于隔天给药.结论 实验研究提示重组人p53腺病毒注射液治疗乳腺癌是有效的,早期连续用药疗效可能会更佳.     

重组人血管内皮抑制素联合放射治疗对lewis肺癌小鼠的作用

背景与目的放疗敏感性与组织中氧含量关系密切,单用重组人血管内皮抑素(rh-endostatin,YH-16,恩度)可能通过血管正常化作用使组织中氧含量增加,这与放疗的敏感性密切相关。本研究将重组人血管内皮抑素与放射治疗联合对lewis肺癌小鼠的肿瘤生长及血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的影响。方法制作lewis肺癌细胞种植肿瘤的动物模型,小鼠随机分为四组:A组为空白对照组,B组为重组人血管内皮抑素组,C组为放疗组,D组为联合组。分别给予相应处理后绘制肿瘤生长曲线,计算抑瘤率,用免疫组化(SP法)测定VEGF的表达及微血管密度。结果经过相应的处理后,各处理组较A组移植瘤的生长速度明显减慢。且各组瘤质量明显低于A组(P<0.05),D组较其它三组降低得更加明显(P<0.05),VEGF的表达及微血管密度与A组相比也有不同程度的降低(P<0.05),其中D组下降最为明显。结论重组人血管内皮抑素和放疗联用对lewis肺癌小鼠有明显肿瘤抑制作用,其机制可能是下调VEGF的表达、抑制新生血管的生成。     

重组人p53腺病毒联合肿瘤坏死因子对乳腺癌Ca761细胞的抑制作用

目的:探讨重组人p53腺病毒注射液(rAd/p53)联合肿瘤坏死因子(TNF)对荷瘤小鼠体内外的乳腺癌细胞Ca761的抑制效果.方法:MTT法,流式细胞术检测细胞凋亡率,观察rAd/p53和TNF对乳腺癌细胞的体外抑制作用及促凋亡作用.建立乳腺癌Ca761细胞小鼠移植瘤模型,瘤内分别注射rAd/p53和TNF及两药联合,观察对肿瘤生长的影响,并对肿瘤组织进行病理学和免疫组化检查.结果:rAd/p53和TNF体内外均可显著抑制乳腺癌Ca761细胞的生长,可以促进乳腺癌细胞Ca761株的凋亡.两药联合对肿瘤细胞具有协同杀伤作用.结论:重组人p53腺病毒注射液(rAd/p53)治疗Ca761乳腺癌细胞移植瘤是有效的,TNF也可抑制其生长,rAd/p53和TNF联合应用有协同作用,为临床乳腺癌的综合治疗提供了依据.     

榄香烯对裸鼠胃癌原位移植瘤血管生成的抑制作用

 目的 观察榄香烯对裸鼠胃癌原位移植瘤生长和血管生成的抑制作用。 方法 采用裸小鼠胃癌原位移植模型,随机分为0.9%氯化钠溶液(NS)组、5-Fu组、榄香烯组和联合组 ,腹腔注射给药。比较各组移植瘤瘤重的差异;免疫组织化学法检测肿瘤微血管密度 (Microvessel Density,MVD) 和VEGF、p53蛋白表达,RT-PCR法检测VEGF mRNA表达。 结果 联合组裸鼠胃癌移植瘤的瘤重显著低于NS组(P＜0.05);榄香烯组、联合组瘤组织MVD、VEGF蛋 白、p53蛋白及VEGF mRNA表达亦明显低于NS组(P＜0.05);各组移植瘤的瘤重与瘤组织MVD呈正 相关(r=0.669,P＜0.01)。 结论 榄香烯能抑制裸鼠胃癌原位移植瘤生长和血管生成,其机制可能与抑制裸鼠胃癌组织VEGF和突 变型p53的表达有关。     

贝伐单抗与重组人血管内皮抑素对人乳腺癌MCF-7细胞裸鼠移植瘤生长的影响

[目的]探讨贝伐单抗、重组人血管内皮抑素对人乳腺癌MCF-7细胞裸鼠移植瘤生长的影响。[方法]建立人乳腺癌裸鼠移植瘤模型,随机分为对照组、低剂量贝伐单抗组、高剂量贝伐单抗组、低剂量莺组人血管内皮抑素组、高剂量重组人血管内皮抑素组、低剂量联合组以及高剂量联合组,用药3周。检测裸鼠体重、移植瘤体积、移植瘤重量,计算抑瘤率。[结果]与对照组相比,低剂量贝伐单抗组、高剂量贝伐单抗组、低剂量联合组、高剂量联合组裸鼠移植瘤生长曲线较平缓,移植瘤重量明显下降（P〈0．01）,抑瘤率分别为67．69％、68．88％、78.32％和79．26％。低剂量联合组与低剂量贝伐单抗组移植瘤重量存在统计学差异（P〈0．05）。低剂量重组人血管内皮抑素组、高剂量重组人血管内皮抑素组移植瘤生长与对照组无统计学差异（P〉0．05）。[结论]贝伐单抗能抑制人乳腺癌MCF-7细胞裸鼠移植瘤生长,低剂量贝伐单抗联合重组人血管内皮抑素能进一步提高抗肿瘤作用。     

重组人干扰素a2b联合顺铂对骨肉瘤的作用及其对VEGF表达的影响

目的探讨重组人干扰素a2b联合顺铂对骨肉瘤的作用效果及其对血管内皮生长因子(VEGF)表达的影响。方法将32只荷瘤裸鼠随机均分为4组:对照组、干扰素a2b组、顺铂组和联合治疗组。分别在接种后第0、5、10、15、20、25、30天记录肿瘤体积变化和抑瘤率;接种后30 d后处死裸鼠,取裸鼠移植瘤测定重量,采用免疫组化法进行CD34标记,用Weidner方法计数微血管密度(MVD);显微镜下检测细胞凋亡情况;ELISA法检测VEGF水平。结果 4组裸鼠成瘤率均为100%。其中,联合治疗组在接种后第10、15、20、25、30天的肿瘤体积显著低于顺铂组和对照组。联合治疗组的瘤体重量显著低于顺铂组和对照组[(0.733±0.054)g vs.(2.326±0.151)g、(2.419±0.166)g],差异均有统计学意义(均P0.05)。联合治疗组MVD值显著低于顺铂组和对照组[(22.86±2.86)vs.(27.45±1.64)、(31.8±3.12)],联合治疗组移植瘤凋亡指数高于顺铂组和对照组[(45.62±4.24)%vs.(9.66±1.15)%、(10.00±1.28)%],差异均有统计学意义(均P0.05)。干扰素a2b组和联合治疗组的VEGF蛋白表达阳性率分别为20.62%、15.45%,低于顺铂组和对照组(59.24%、68.37%),差异有统计学意义(P0.05)。结论重组人干扰素a2b联合顺铂注射能特异性抑制骨肉瘤裸鼠VEGF的表达、血管生成和肿瘤生长,并能促进肿瘤细胞凋亡。     

放疗联合周剂量重组人血管内皮抑素对A549肺腺癌裸鼠移植瘤生长抑制作用的研究

目的:研究放疗联合周剂量重组人血管内皮抑素(recombinant human endostatin, rh-Endostatin)对肺腺癌A549裸鼠移植瘤生长的抑制作用.方法: 40 只裸鼠A549细胞移植瘤模型随机分成4组:空白对照组、rh-Endostatin治疗组、放射治疗组和放疗联合rh-Endostatin治疗组,观察并绘制肿瘤生长曲线图,计算肿瘤体积抑制率;肿瘤组织行常规HE病理学检查,免疫组织化学法检测肿瘤组织中微血管内皮CD31的表达及肿瘤微血管密度(microvessel density, MVD)的变化;采用免疫组织化学及Western印迹法检测血管内皮生长因子(vascular endothelial growth factor, VEGF)的表达;TUNEL法检测肿瘤细胞的凋亡. 结果:治疗第8天起,放疗联合rh-Endostatin治疗组的肿瘤体积与对照组相比,差异有统计学意义(P＜0.05).15 d后,rh-Endostatin治疗组、放射治疗组和放疗联合rh-Endostatin治疗组小鼠的肿瘤体积的抑制率依次为68.35%、90.78%和106.56%; rh-Endostatin治疗组小鼠的MVD较放射治疗组下降明显(P＜0.05);但rh-Endostatin治疗组与对照组、放疗联合rh-Endostati治疗组及放射治疗组相比,VEGF的改变差异均无统计学意义;放疗联合rh-Endostatin治疗组细胞凋亡明显.结论:放疗联合周剂量rh-Endostatin能抑制肿瘤的生长,较早诱导肿瘤退缩,可能与减少放射治疗后肿瘤血管再生及增加肿瘤细胞和内皮细胞的凋亡相关;各治疗组小鼠均未出现急性不良反应,因此该方法具有短疗程的优势,可用于临床推广.     

重组人血管内皮抑素抑制Namalwa淋巴瘤裸鼠移植瘤生长的实验研究

目的:探讨重组人血管内皮抑素对Namalwa淋巴瘤裸鼠移植瘤生长的抑制作用.方法:建立Namalwa淋巴瘤裸鼠移植瘤模型.从Namalwa淋巴瘤细胞种植后第11天开始给予腹腔注射2.5、5和10 mg/kg重组人血管内皮抑素,以腹腔注射75 mg/kg环磷酰胺作为阳性对照组,腹腔注射0.9%氯化钠溶液作为阴性对照组.比较各组相对肿瘤体积(relative tumor volume,RTV),计算相对肿瘤增殖率.结果:药物干预8 d后,环磷酰胺组RTV小于阴性对照组;干预12 d后,5 mg/kg重组人血管内皮抑素组移植瘤RTV小于阴性对照组(P＜0.05),但大于环磷酰胺组(P＜0.05);而2.5 和10 mg/kg组与阴性对照组之间的差异无统计学意义.2.5、5和10 mg/kg重组人血管内皮抑素组和环磷酰胺组的肿瘤相对增殖率分别为87.4%、64.2%、85.9%和39.2%.结论:重组人血管内皮抑素可抑制Namalwa淋巴瘤裸鼠移植瘤生长,其抑瘤作用与合适的剂量和足够的疗程相关.     

Lack of association of human polyomaviruses with human brain tumors

Summary Brain tumors, primarily glioblastoma multiforme, were examined for the presence of BKV DNA. DNAs extracted from 33 fresh-frozen tumors and from an additional 47 paraffin-embedded tumor tissues were tested for BKV sequences using two different primer pairs. One primer pair amplified highly conserved sequences in the early region coding for BKV and JCV T antigens. The other primer pair amplified the regulatory region of BKV-IR, a variant previously associated with human brain tumors. None of the tumors were positive for BKV or JCV DNA.     

Detection of human papillomavirus DNA in biopsies of human oral tissue

We have employed molecular probes produced from DNA fragments of human papillomavirus, cloned into prokaryotic vectors, to detect virus nucleic acid sequences in extracts of human oral tissues. The study was conducted with duplicate coded snap-frozen tissue biopsies from which frozen sections had been taken to accurately assess the pathology of each particular sample. The results show that a large proportion of the oral biopsies contained DNA which hybridized to the viral DNA probes, even under conditions of high stringency. The presence of virus did not correlate with neoplasia in the tissues examined, but HPV like sequences were found in a high proportion (80%) of biopsies taken from areas of keratosis and lichen planus and also in 41 to 46% of normal and tumour tissues.     

Inhibition of human immunodeficiency virus infection of human lymphocytes by gangliosides

As the first step in human immunodeficiency virus (HIV) infection, HIV binds to CD4 molecules on the surface of human T lymphocytes. Expression of CD4 by the cells is remarkably modulated by exogenously added gangliosides, which are normal components of the surface of animal cells. We report here that these physiological molecules also clearly inhibited HIV infection of lymphocytes in vitro in a dose-dependent manner through the selective modulation of CD4 from the cell surface. This raises the possibility of in vivo application of gangliosides as a new strategy for treating acquired immunodeficiency syndrome.     

Biologic effect of human hepatocyte growth-factor on human gastric-carcinoma cell-lines

Toxicity of polyphenols against rat 3Y1 fibroblasts and the cells transformed by human adenovirus (Ad12-3Y1), its EIA gene (EIA-3Y1), or simian virus 40 (SV-3Y1) was examined. Among the diphenol compounds examined, pyrocatechol (o-diphenol) and hydroquinone (p-diphenol) showed selective toxicity against Ad12-3Y1 and EIA-3Y1 cells, while resorcinol (m-diphenol) showed a much weaker non-specific toxicity against these cells. Another o-diphenol (dopamine) and triphenols (gallic acid and pyrogallol) were less toxic but showed selective toxicity. At lower concentrations where they were not toxic, all polyphenols attenuated toxicity of phosphatidylcholine against EIA-3Y1 cells. Among antioxidants examined, ascorbic acid reduced the toxicity of pyrocatechol, but alpha-tocopherol and butyrated hydroxytoluene did not. Oxidation of pyrocatechol was not enhanced in the presence of 3Y1 or EIA-3Y1 cells and their homogenates. These results suggest that the selective toxicity of polyphenols against Ad12-3Y1 and E1A-3Y1 cells is not related to their oxidation velocity but other factors such as the activity of active oxygen-scavenging enzymes.     

Ectopic production of human chorionic gonadotrophin by human breast tumours

The incidence of tumours ectopically producing the human chorionic gonadotrophins was studied in patients with breast cancer. Specific radioimmunoassay of subunits of HCG was utilized. Nine out of 65 patients with carcinoma of breast showed the presence of circulating HCG. Patients with other pathological conditions of breast tissue did not show any evidence of circulating HCG.     

重组人内皮抑素对荷人肺腺癌裸鼠肿瘤微环境的调节作用

目的研究重组人内皮抑素对荷人肺腺癌裸鼠肿瘤微环境的调节作用。方法构建人肺腺癌裸鼠模型,重组人内皮抑素5mg／kg皮下给药14天。采用荧光素（TRITC—Dextran）示踪法,荧光显微镜观察重组人内皮抑素处理后不同时间点荧光素在肿瘤中的分布,并计算肿瘤实质内部荧光素不能弥散到的面积相对于总面积的比例。结果给予处理14天后,对照组肿瘤内部血液弥散减少（低弥散面积占总面积的10．01％±4．50％）,相对于第7和第21天显著增高（分别为1．49％±1．14％和2．90％±2．02％）。重组人内皮抑素能够在治疗的初始阶段抑制血液的弥散（低弥散面积占总面积的16．85％±4．03％）,但第14天后这种抑制作用解除,肿瘤内部血液弥散反而增加（0．54％±0．32％,P=0．000）,到第21天时则两组没有明显差别。讨论重组人血管内皮抑素能够暂时性的抑制肿瘤血管,此外重组人内皮抑素还能够调节肿瘤的内环境,在一段时间内增加了肿瘤内部血液弥散。     

Effect of recombinant human interleukin 4 on human monocyte activity

Recombinant human interleukin 4 (rhuIL-4), a lymphokine that reportedly stimulates tumoricidal activity in mouse macrophages, is currently undergoing clinical studies to determine its efficacy in the treatment of cancer. IL-4 is known to participate with other cytokines to regulate growth and differentiation of various hematopoietic cells as well as modulate the immune response. Little is known about the effect of rhuIL-4 on human monocyte tumoricidal activity. The purpose of these studies was to examine the effect of rhuIL-4 on human peripheral blood monocytes. Peripheral blood monocytes isolated from normal donors failed to demonstrate tumoricidal activity or interleukin 1 secretion after treatment with rhuIL-4 in vitro. Furthermore, monocyte-mediated cytotoxicity induced by recombinant human gamma-interferon plus muramyl dipeptide was suppressed in a dose-dependent manner by rhuIL-4. This reduction in cytotoxicity corresponded to a reduction in IL-1 production and secretion. Further investigation of rhuIL-4 and its role in the cytokine network is necessary for the development of effective immunotherapy in cancer patients.     

Involvement of human T lymphotropic virus type I in human neoplasia

Of 211 human tissues analyzed by PCR-SSCP and direct sequencing for HTLV-I genes, 165 (78.2%) gave positive signals. Signals detected in family members of adult T cell leukemia (ATL) patients showed mutations distinct from those in ATL. HTLV-I gene sequences showed variability in same persons examined at different times. Lymphocytes with prototypic HTLV-I genes showed higher sister chromatid exchange following MNNG treatment than those without. Nasopharyngeal carcinomas carrying both HTLV-I and Epstein-Barr virus genes showed markedly abnormal gene product expression. HTLV-I genes in human DNA may cause deregulation of host cell genes upon exposure to carcinogens including viruses.