Increased expression of the glucose-regulated gene encoding the Mr 78,000 glucose-regulated protein in chemically and radiation-transformed C3H 10T1/2 mouse embryo cells.
Increased expression of the glucose-regulated gene encoding the Mr 78,000 glucose-regulated protein in chemically and radiation-transformed C3H 10T1/2 mouse embryo cells.
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发表时间：
1987-12
期刊：
影响因子：
11.2
通讯作者：
S. Patierno;J. Tuscano;K. Kim;J. Landolph;A. Lee
中科院分区：
文献类型：
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作者：
S. Patierno;J. Tuscano;K. Kim;J. Landolph;A. Lee
在非转化和化学和辐射转化的C3 H 10 T1/2 C1 8小鼠胚胎细胞中检测了编码Mr 78，000葡萄糖调节蛋白（GRP 78）的基因的表达。当细胞在含有10%胎牛血清的完全培养基中生长时，与未转化的10 T1/2克隆8细胞（Cl 8）相比，在3-甲基胆蒽（MCA;克隆15和16）-、博来霉素（Bleo 1）-和紫外线（UV-C3）-转化的细胞系中，在相似的细胞数量和生长期，GRP 78 mRNA增加了4- 9倍。增加稳态水平的GRP 78蛋白在MCA Cl 15相比，Cl 8细胞证实了二维凝胶电泳。在这些条件下，转化的MCA C115表现出增加的GRP 78 RNA后24小时内加入新鲜的含葡萄糖的培养基，而非转化的C18细胞没有增加该基因的表达，即使在条件培养基中培养5天后。C18和MCA C115在无葡萄糖培养基中孵育16小时分别引起GRP 78 RNA的3倍和15倍诱导。此外，化学转化的细胞对葡萄糖剥夺高度敏感，并通过聚集和脱离基质来响应。C18细胞对葡萄糖剥夺无此敏感性.这些结果扩展了早期的报告病毒转化的细胞，包括化学和辐射转化的细胞和扩大早期的报告，包括mRNA表达和2维凝胶电泳的GRP 78蛋白。
Expression of the gene coding for the Mr 78,000 glucose-regulated protein (GRP78) was examined in nontransformed and chemically and radiation-transformed C3H 10T1/2 Cl 8 mouse embryo cells. When cells were grown in complete medium with 10% fetal bovine serum, GRP78 mRNA was increased 4- to 9-fold in 3-methylcholanthrene (MCA; Clones 15 and 16)-, bleomycin (Bleo 1)-, and ultraviolet light (UV-C3)-transformed cell lines compared to nontransformed 10T1/2 clone 8 cells (Cl 8) at similar cell number and growth phase. Increased steady-state levels of GRP78 protein in MCA Cl 15 compared to Cl 8 cells were confirmed by 2-dimensional gel electrophoresis. Under these conditions transformed MCA Cl 15 exhibited increased GRP78 RNA within 24 h after addition of fresh glucose-containing medium, whereas nontransformed Cl 8 cells did not increase expression of this gene even after 5 days of culture in conditioned medium. Incubation of Cl 8 and MCA Cl 15 in glucose-free medium for 16 h caused a 3- and 15-fold induction of GRP78 RNA, respectively. In addition, chemically transformed cells were highly sensitive to glucose deprivation and responded by rounding up and detaching from the substratum. Cl 8 cells exhibited no such sensitivity to glucose deprivation. These results extend earlier reports on virally transformed cells to include chemically and radiation-transformed cells and expand earlier reports to include mRNA expression and 2-dimensional gel electrophoresis of GRP78 protein.