[Studies on the target cells and molecules with sodium valproate induced differential of human glioma cells].
OBJECTIVE
To investigate the target cells and molecules with sodium valproate induced differentiation of human glioma cells.


METHODS
Nude mice bearing human glioma xenogenic graft subcutaneously were treated with sodium valproate. The expressions of HDAC1 and Tob genes of xenografts were analyzed with semiquantitative RT-PCR. The CD133+ cells (BTSCs) were isolated from glioma specimens by immunomagnetic sorting, and cultured in the medium containing FCS or in the serum-free medium supplemented with growth factors, respectively, followed by treatment with sodium valproate in vitro for 21 days. The cell surface markers were detected with flow cytometry and confocal microscopy.


RESULTS
Sodium valproate inhibited the growth of subcutaneous xenografs bearing on nude mice (P<0.05), and up-regulated the HDAC1 expression (P<0.01), down-regulated the Tob expression (P<0.05). The cell surface markers of BTSCs were detected by flow cytometry after sodium valproate treatment for 21 days. In the FCS group, the GFAP or beta-tubulin III positive cells increased significantly (P<0.01), but in the growth factor group, no statistical differences were observed in the GFAP or beta-tubulin III expression (P>0.05). The results of confocal microscopy indicated that the GFAP+ or beta-tubulin III+ cells coexpressed with Nestin.


CONCLUSIONS
HDAC1 and Tob genes were the potential target molecules in reversion of the differential inhibition of human glioma cells with sodium valproate. The BTSCs undergoing the processes of differentiation were the target cells for sodium valproate.
Wang
Ai-dong
AD
Department of Neurosurgery and Brain Tumor Laboratory, the Second Affiliated Hospital of Suzhou University, Suzhou 215004, China.
Ji
Xiao-yan
XY
Huang
Qiang
Q
Wang
Chong-ren
CR
Dong
Jun
J
Lan
Qing
Q
D000071916:AC133 Antigen; D000199:Actins / Q000032:analysis; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D002454:Cell Differentiation / Q000187:drug effects*; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D005910:Glioma / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D006023:Glycoproteins / Q000032:analysis; D006655:Histone Deacetylases / Q000235:genetics; D006801:Humans; D007381:Intermediate Filament Proteins / Q000032:analysis; D051379:Mice; D008819:Mice, Nude; D018613:Microscopy, Confocal; D009419:Nerve Tissue Proteins / Q000032:analysis; D064231:Nestin; D010455:Peptides / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D014635:Valproic Acid / Q000494:pharmacology*; D023041:Xenograft Model Antitumor Assays / Q000379:methods
D000071916:AC133 Antigen; D000199:Actins; D015703:Antigens, CD; D005904:Glial Fibrillary Acidic Protein; D006023:Glycoproteins; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D025521:Tumor Suppressor Proteins; D014635:Valproic Acid; D006655:Histone Deacetylases