Change of morphology and cytoskeletal protein gene expression during dibutyryl cAMP-induced differentiation in C6 glioma cells.
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.
Hu
Weiwei
W
Department of Pharmacology, School of Medicine, Zhejiang University, Hangzhou 310058, China. originoflife78@hotmail.com
Onuma
Takeshi
T
Birukawa
Naoko
N
Abe
Masashi
M
Ito
Etsuro
E
Chen
Zhong
Z
Urano
Akihisa
A
D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D048430:Cell Shape / Q000187:drug effects*; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D005910:Glioma / Q000473:pathology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D014404:Tubulin / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000199:Actins; D003598:Cytoskeletal Proteins; D012333:RNA, Messenger; D014404:Tubulin; D003994:Bucladesine