Change of morphology and cytoskeletal protein gene expression during dibutyryl cAMP-induced differentiation in C6 glioma cells
Change of morphology and cytoskeletal protein gene expression during dibutyryl cAMP-induced differentiation in C6 glioma cells
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DOI：
10.1007/s10571-007-9229-y
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发表时间：
2008-06-01
影响因子：
4
通讯作者：
Urano, Akihisa
中科院分区：
文献类型：
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作者：
Hu, Weiwei;Onuma, Takeshi;Urano, Akihisa
C6胶质瘤细胞内cAMP水平升高诱导星形胶质细胞样分化的形态学改变这种变化可能伴随着细胞骨架蛋白基因的表达。因此，我们分析了用二丁酰cAMP（dbcAMP）处理后的形态学变化，然后通过定量实时聚合酶链反应评估细胞骨架蛋白基因的表达。细胞数保持不变，孵育后与1 mM的dbcAMP在培养基中补充有0.1%胎牛血清（FBS），而过程的数量和长度增加，当与那些在培养基中孵育的细胞补充有0.1%或10%FBS只。在含10%FBS的DMEM中，C6细胞中β-肌动蛋白、γ-肌动蛋白和β-微管蛋白mRNA的量在早期增殖期间增加，但α-微管蛋白mRNA不增加。与10%FBS培养的细胞相比，0.1%FBS或1 mM dbcAMP/0.1%FBS培养的细胞骨架蛋白基因表达降低。这些结果表明，在正常培养条件下，C6细胞在早期增殖过程中，细胞骨架蛋白基因的表达，除了α-微管蛋白，增加，而在分化或分化的C6胶质瘤细胞，cAMP诱导的形态学变化并不伴随着细胞骨架蛋白，如肌动蛋白和微管蛋白的基因表达的升高。
Elevation of the intracellular cAMP level induces morphological changes of astrocyte-like differentiation in C6 glioma cells. Such changes may be accompanied with expression of cytoskeletal protein genes. We therefore analyzed morphological changes after a treatment with dibutyryl cAMP (dbcAMP) and then assessed the expression of cytoskeletal protein genes by a quantitative real-time polymerase chain reaction. The cell number remained unaltered upon incubation with 1 mM dbcAMP in medium supplemented with 0.1% fetal bovine serum (FBS), whereas the number and lengths of processes increased, when compared with those of cells incubated in medium supplemented with 0.1% or 10% FBS only. The amounts of beta-actin, gamma-actin, and beta-tubulin mRNAs in C6 cells, but not alpha-tubulin mRNA, increased during the early proliferation in DMEM containing 10% FBS. The expression of cytoskeletal protein genes decreased when incubated with 0.1% FBS or 1 mM dbcAMP in 0.1% FBS, compared with those of cells cultured in 10% FBS. These results indicated that, during the early proliferation in normal culture condition, the expression of cytoskeletal protein genes in C6 cells, except alpha-tubulin, increased, while in differentiating or differentiated C6 glioma cells, cAMP-induced morphological changes were not accompanied with elevation of gene expression for cytoskeletal proteins, such as actin and tubulin.