Thomopoulos G N, Kyurkchiev S, Perbal B
Aristotle University, School of Sciences, Department of Biology, Thessaloniki, Greece.
J Submicrosc Cytol Pathol. 2001 Jul;33(3):251-60.
In this study we present biochemical and immunocytochemical results on NOVH protein secretion and localization in NCI-H295R cells, as well as results on the ultrastructural characteristics of NCI-H295R cells. NCI-H295R cells were characterized by small quantities of rough and smooth endoplasmic reticulum, many free ribosomes, large nuclei with prominent nucleoli, numerous elongated mitochondria, a few Golgi complexes, and a small number of lipid droplets. Large numbers of coated pits and coated vesicles were present, but no secretory granules or exocytotic profiles were seen. Best ultrastructural preservation of NCI-H295R cells was achieved when fixation was done directly on the culture dishes and the cells were detached by scraping. Our biochemical results showed that NCI-H295R cells secreted large amounts of NOVH protein. The immunocytochemical localization of NOVH protein showed that the protein was localized in the cytoplasm, the plasma membrane and the nuclear envelope. This localization pattern, along with the ultrastructural and biochemical findings raise interesting questions on the function(s) and the mode of secretion of NOVH protein.
在本研究中，我们展示了关于NOVH蛋白在NCI-H295R细胞中的分泌及定位的生化和免疫细胞化学结果，以及NCI-H295R细胞超微结构特征的结果。NCI-H295R细胞的特征为粗面和滑面内质网数量少、游离核糖体多、细胞核大且核仁突出、线粒体众多且呈细长形、高尔基体复合体少以及脂滴数量少。存在大量有被小窝和有被小泡，但未见分泌颗粒或胞吐结构。当直接在培养皿上进行固定并通过刮擦使细胞脱离时，可实现NCI-H295R细胞最佳的超微结构保存。我们的生化结果表明，NCI-H295R细胞分泌大量的NOVH蛋白。NOVH蛋白的免疫细胞化学定位显示，该蛋白定位于细胞质、质膜和核膜。这种定位模式，连同超微结构和生化研究结果，引发了关于NOVH蛋白的功能和分泌方式的有趣问题。
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