17—β雌二醇对原代人脐静脉内皮细胞一氧化氮合酶基因表？…

目的：阐明雌二醇对内皮细胞一氧化氮（ＮＯ）生成及一氧化氮化合酶（ｃＮＯＳ）基因表达的影响，方法；观察雌二醇作用于原代人脐静脉内皮细胞（ＨＵＶＥＣ）一定时间（１－２４ｈ）后的ＮＯ产量，采用Ｇｒｅｉｓｓ反应测定代谢产物ＮＯ２的量及Ｎｏｒｔｈｅｒｎ杂交技术分析雌二醇作用于ＨＵＶＥＣ１８ｈｃＮＯＳｍＲＮＡ的表达，结果：１００，３０，１０及１ｎｍｏｌ．Ｌ＾－１的雌二醇作用下，ＨＵＶＥＣ分别于１，４，１２及１     

ADENOVIRUS-MEDIATED EXPRESSION OF PEX, A NONCATALYTIC FRAGMENT OF MATRIX METALLOPROTEINASE-2, AND IT＇S INHIBITION ON ANGIOGENESIS AND TUMOR GROWTH

Objective： To develop an adenovirus system to deliver biologically active peptides or proteins such as angiogenesis inhibitors in vivo for the treatment of cancer. Methods： DNA recombination techniques were employed to construct adenovirus shuttle vector, in which angiogenesis inhibitor was put downstream of rat growth hormone signal peptide, and the C-terminal was the myc-epitope 10-amino-acid peptide for the following up of the protein. Adenovirus was made using the bacteria recombination method. We tested this system using an angiogenesis inhibitor chick MMP-2 C-terminal hemopexin-like fragment （PEX） in Sarcoma 180 （S-180） bearing Kunming mice. The anti-angiogenie effect was performed by chick chorioallantoic membrane assay. Results： PEX was readily secreted outside human stomach carcinoma BGC823 cells as demonstrated by immunofluorcscent staining and western blot infected by adenovirus with rat growth hormone signal peptide （E-T-rGH-PEX）. However, without signal pcptide （E-T-PEX）, PEX was expressed and localized in the cytoplasm of the infected cells, and formed large aggregates, which suggested that PEX was insoluble. The adenovirus E-T-rGH-PEX could inhibit angiogenesis, while E-T-rGH-PEX not. The adenoviruses of E-T-rGH-PEX inhibited the growth of S-180 tumor significantly compared with the empty virus control group E-T （P=0.026） and without signal peptide group E-T-PEX （P=0.006） respectively, while E-T-PEX had little effect. Conelusion： These results suggest that this adenoviral system is likely to be used in the gene therapy of cancer to deliver angiogenesis inhibitors.     

老年医学与肿瘤学

老年医学与肿瘤学林仲翔一年一度的美国国际Ｇｏｒｄｏｎ学术会议１９９５年度在意大利Ｖｏｌｔｅｒｒａ于５月５～６日召开了以衰老研究为主题的卫星会，专题为＂对老年的干预＂。会上美国密西根大学老年医学研究所病理系ＲＡＭｉｌｌｅｒ博士就老年医学与肿瘤学问题作了...     

自发永生性转化人胚肺上皮细胞系HLEC的建立及鉴定

目的对源自人胚肺组织原代培养、形态似上皮类型的细胞系——人肺上皮细胞(HLEC)的生物学特性进行鉴定,为相关研究和应用提供模型和依据。方法分别采用相差显微镜下观察、细胞计数、β-半乳糖苷酶染色、免疫荧光细胞化学。人Alu序列PCR扩增、染色体计数、软琼脂集落形成实验和裸鼠致瘤实验对HLEC的形态学特征、生长特性、衰老状态、上皮细胞标志物、来源、细胞遗传学特点和恶性转化等生物学特性进行研究鉴定。结果HLEC角蛋白和上皮型细胞间黏附分子钙黏着素E-cadherin免疫荧光染色阳性,染色体众数在41～44,人Alu序列PCR扩增结果阳性,β-半乳糖苷酶阳性细胞的比例未见随细胞代数的增加而升高,软琼脂和裸鼠体内均不生长,微丝及黏着斑结构正常。结论HLEC细胞具有永生化细胞特征、属相对正常人肺上皮亚二倍体细胞,可用于肺癌癌变机理研究和癌细胞的相对正常对照。     

横纹肌肉瘤逆转分化细胞模型研究

作者观察到人横纹肌肉瘤ＲＤ系细胞的恶性表型明显，细胞增殖快、增殖与分化的关系异常、肌分化阻断、细胞通讯功能缺陷、间隙连接基因Ｃｘ４３转录抑制。用单细胞克隆分离技术和Ｃｘ４－３ｃＤＮＡ转染方法获得ＲＤ细胞亚系，从中选出表型差别明显的ＲＤＬ６、ＲＤＬ３和ＲＤＬ６／Ｃ－４三个亚系进行生物学特性分析比较。结果表明三个亚系在细胞增殖速度、增殖与分化偶联关系、肌分化程度、ｃｘ４３基因功能性表达和原癌基因ｃ－ｍｙｃ、Ｈａ－ｒａｓ、ｃ－ｍｅｔ表达方面有明显差别，可以作为研究ＲＤ逆转分化的细胞模型。作者对ＲＤ逆转分化的阶段性标志、意义及有关检查方法的应用价值进行了讨论。     

三磷酸腺苷促进巨噬细胞吞噬功能的实验研究

巨噬细胞通过其吞噬功能可清除炎症及肿瘤区域的病原体及坏死组织,在防御体系中发挥重要作用,当前肿瘤治疗常用的放疗化疗手段是引起细胞的凋亡,所形成的凋亡小体也依赖于巨噬细胞吞噬功能,但放疗化疗又同时造成巨噬细胞吞噬功能的降低,因此,研究吞噬细胞吞噬功能的提高有重要理论和实际意义.     

高转移人肺癌表型去恶化研究─一细胞通讯、细胞骨架和癌基因、抑癌基因的相关变化

高转移人肺巨细胞癌ＰＧ细胞恶性表型明显。以人胚肺细胞为对照，荧光染料标记示踪法表明，ＰＧ的间隙连接通讯功能缺陷；免疫荧光染色显示微管解聚；荧光鬼笔环肽染色显示微丝网架破坏。分子杂交结果表明，ＰＧ细胞有ｃ－ｍｙｃＤＮＡ扩增和ｃ－Ｈａ－ｒａｓ、ｃ－ｍｙｃ基因高表达，抑癌基因Ｐ５３ｍＲＮＡ水平低于正常。用钙调蛋白拮抗剂ＣＤＺ（１００～２００ｎｍｏｌ／Ｌ）和中药Ｌ２（３～１３ｍｇ／ｍｌ）分别处理ＰＧ，细胞增殖受到抑制，表型向正常分化，细胞通讯功能再现，微管网架恢复。但只有Ｌ２能促进微丝骨架恢复，抑制软琼脂内的集落形成，并使ｃ－ｍｙｃ扩增减弱和ｍＲＮＡ水平下降，Ｐ５３表达升高。     

α—catenin／GST融合蛋白的原核表达及其多克隆抗体制备

目的 制备α-catenin多克隆抗体,为深入研究其功能和探讨其与肿瘤等疾病的相关性提供工具。方法 PCR扩增编码人α-catenin C－端455个氨基酸的DNA片段,DNA重组入原核表达质粒pGEX-4T-3,转化大肠杆菌JM109菌株,异丙基β－D硫代半乳糖苷（IPTG）诱导产生GST／α-catenin融合蛋白。GST－Sepharose亲和层析纯化可溶性融合蛋白,免疫新西兰白兔,制备抗血清。通过ELISA、免疫荧光细胞化学和SP免疫组织化学法鉴定血清特异性和效价。结果 成功地构建了pGEX-4T-3/α-Cat表达载体,转化JM109后可高效表达融合蛋白GST－α-catenin（α－Cat）。免疫兔产生的α-Cat多抗可特异检测人体组织和细胞中α-Cat表达及定位情况。结论 α-catenin抗体效价高,特异性强,适合免疫组织化学、ELISA等检测应用。     

ACTIVATION OF NUCLEOLAR rRNA GENE TRANSCRIPTION IN LYMPHOCYTES BY TUMOR PROMOTERS: APPLIES STUDIES WITH IN SITU HYBRIDIZATION DETECTED BY FLUORESCENT METHOD

The distribution of rDNA was visualized in interphase nuclei of tumor promoter treated human lymphocytes in comparison with the mltogen Phytoheamagglutinin (PHA) effects by using an in situ hybridization fluorescent method. The procedure involves biotinylated rDNA as the probe and FITC-avidin as detection system. Silver (Ag) staining was used to visualize nucleoli. In the interphase nuclei of most of the nonstimulated control lymphocytes, only one small Ag-stained nucleolus could be seen. The in situ hybridization, however, revealed one to several agglomerations of rDNA fluorescent spots. With tumor promoting herb extract WCE (40 μg/ml) or TPA (60 ng/ ml) treatment, the Interphase nucleoli increased slightly in number with the morphology alteration into larger, reticular or compact granular types. Ag-stained particles also increased in number. The number of the in situ hybridization rDNA fluorescent spots and dots increased markedly and largereticulate formations of numerous rDNA spots were seen. This     

三磷酸腺苷对胃癌细胞Na~＋－K~＋－三磷酸腺苷酶活性和c－myc基因表达的影

三磷酸腺苷对胃癌细胞Ｎａ￣＋－Ｋ￣＋－三磷酸腺苷酶活性和ｃ－ｍｙｃ基因表达的影响吕桂芝，林仲翔，孟松娘，高燕，周立新，徐光炜三磷酸腺苷（ＡＴＰ）对多种癌细胞的增殖均具有明显的抑制作用［１］，本实验应用电镜细胞化学和免疫细胞化学方法证实了ＡＴＰ抑制人胃...