人食管癌细胞与正常食管上皮细胞内微管的免疫荧光细胞化学研究

本文利用抗微管蛋白抗体间接免疫荧光细胞化学技术,观察到人食管癌ECa109细胞的微管分布与正常食管上皮细胞有明显差别。原代培养的人正常食管上皮细胞胞质内有间期的纤细的微管结构(CMTC)和分裂期的纺锤体微管。食管癌ECa109细胞在间期的胞质内缺乏微管,但分裂期的纺锤体微管与正常细胞相同。本文对癌细胞间期胞质缺乏微管的现象进行了讨论,认为可能涉及微管组装机制的缺陷。     

IMMUNOFLUORESCENT STUDIES ON MICROTUBULES OF NORMAL AND MALIGNANT HUMAN ESOPHAGEAL CELLS

Anti-tubulin antibody indirect immunofluorescenee microscopy is used in this report. Difference of intracellular microtubule fluorescent staining patterns between normal and malignant human esophageal cells has been observed.Primarily cultured human esophageal epithelial cells display an elaborate array of fluorescent cytoplasmic microtubule filaments, the cytoplasmic microtubule complex(CMTC), in interphase cells and mitotic spindle microtubules in mitotic cells. Malignant esophageal cells from human esophageal carcinoma, ECa109 cells, contain very few cytoplasmic microtubules in interphase cells, but there is no difference of mitotic spindle microtubule fluorescence between normal and malignant cells.The results presented in this report suggest that the diminution of interphase cytoplasmic mierotubules in tumor cells is probably due to the deficiency of microtubule organizing mechanism in interphase tumor cells.     

大鼠神经胶质瘤细胞间隙连接通讯——代谢协同现象及其抑制剂效应的研究

本文用大鼠神经胶质瘤P_(98)F_(47)细胞系建立代谢协同试验(Metabolic Cooperation Test,MCT)体系研究细胞的间隙连接通讯(Gap Junctional Intercellular Communication,GJIC)功能。本实验证明P_(98)F_(47)瘤细胞与瘤细胞之间,瘤细胞与正常神经胶质细胞之间都有代谢协同能力,代表细胞间隙连接通讯功能。不同的促癌剂对P_(98)F_(47)细胞的代谢协同作用有不同的抑制效应。文中用~3H-脲嘧啶标记示踪法进一步证明P_(98)F_(47)细胞的间隙连接通讯功能。对这个试验体系在研究和检测神经系统促癌剂和神经毒素中的效能及意义进行了讨论和评价。     

GAP JUNCTIONAL INTERCELLULAR COMMUNICATION (GJIC) IN RAT GLIOMA CELLS — CHARACTERIZATIONS TO DETECT INHIBITORS OF METABOLIC COOPERATION

Gap junctional intercellular communication (GJIC) as measured by metabolic cooperation was examined in a rat glioma cell line P_(98)F_(47). X-ray induced mutants of P_(98)F_(47) cells were grown in 6-thioguanine selective medium (6TG medium) to separate 6TG-resistant HGPRT- mutant cells (6TG~r). By co-culturing 200 6TG~r cells with varied high densities of the wild type 6TG-sensitive cells (6TG~s), it was found that the recovery of 6TG~r cells depended on the density of 6TG~s cells. Higher densities of 6TG~s cells reduced the recovery of 6TG~r cells. These results demonstrate the ability of P_(98)F_(47) cells to perform metabolic cooperation which is indicative of GJIC. When metabolic cooperation was inhibited, increased recovery of 6TG~r cells was observed. Presented results also demonstrate metabolic cooperation between P_(98)F_(47) glioma cells and normal rat glial cells. Effect of tumor promoting chemicals on metabolic cooperation of P_(98)F_(47) cells was studied.~3H-uridine nucleot de echnique was     

连接蛋白基因表达与横纹肌肉瘤分化关系研究

报道了用间隙连接蛋白Cx43基因探针斜缝杂交,Cx43和肌专一蛋白免疫荧光染色和罗氏黄荧光传输法观察到人横纹肌肉瘤(RD细胞系)的分化异常,细胞通讯缺陷,Cx43表达下降。单细胞克隆亚系RDL6增殖活跃,分化差,细胞通讯缺陷,Cx43表达很低;另一亚系RDL3增殖慢,有明显肌分化,通讯功能强,Cx43及其mRNA的表达近似正常成肌细胞。Cx43 cDNA+neo基因转染RDL6得到Cx43稳定表达的克隆RDL6/C-4,其通讯功能恢复,增殖抑制,成肌性分化增强。本结果证明RD细胞分化阻断与Cx43降表达有关。Cx43的功能性表达不仅抑制瘤细胞增殖,而且与瘤细胞向正常分化有正相关性。