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利用核仁蛋白B23的单克隆抗体以及间接免疫荧光和蛋白质免疫印迹方法,研究了血清饥饿的NIH3T3成纤维细胞受血清诱导重新进入增殖周期后,核仁蛋白B23含量的变化,以及有丝分裂期和间期NIH3T3细胞中B23蛋白含量的差异。结果发现,生长至汇合状态的细胞在无血清培养液中培养24h后,检测到的B23蛋白含量较低,加入10％小牛血清能够引起细胞内B23蛋白含量显著增加,而且在加入血清6h后就已经发生。结果还显示,用微管抑制剂秋水仙酰胺阻断得到的有丝分裂期细胞中B23蛋白含量明显高于间期细胞。以上结果提示,核仁蛋白B23在血清诱导NIH3T3细胞增殖的早期阶段以及细胞由G_2期进入M期时均可能起着一定的调节作用。
Using the monoclonal antibody against nucleocapsid protein B23 and indirect immunofluorescence and Western blotting, the changes of the content of nucleolar protein B23 in serum-starved NIH3T3 fibroblasts after reintroduction into the proliferative cycle induced by serum, Differences of B23 protein content in NIH3T3 cells in The results showed that the B23 protein content of cells cultured in confluent state after incubation for 24h in serum-free culture medium was lower. Adding 10% bovine serum could significantly increase the content of B23 protein in cells, and after adding serum for 6h It happened already. The results also showed that B23 protein was significantly higher in mitotic cells than that in interphase cells, which was blocked by colchicine, a microtubule inhibitor. The above results suggest that nucleolar protein B23 may play a regulatory role in the early stage of NIH3T3 cell proliferation induced by serum and the cell phase from G2 phase to M phase.