Transcription analysis of TIMP-1 and NM23-H1 genes in glioma cell invasion.
OBJECTIVE
To evaluate using transcription analysis the presence and importance of two genes: NM23-H1 and TIMP-1 on control of tumor cell invasion in diffuse astrocytomas (WHO II) and glioblastoma multiforme (WHO IV).


METHODS
Northern blot analysis of NM23-H1 and TIMP-1 was performed. Eight diffuse astrocytomas and 19 glioblastomas (WHO IV) were analyzed to determine if TIMP-1 and NM23-H1 were candidates to inhibition of tumor cell invasion quantitated RNA levels. The samples were collected directly from operating room. Total cellular RNA was extracted from frozen tissue samples using guanidinium-isothiocyanate and cesium chloride gradients. Total RNA (10 mg per sample) from tumor tissue were size fractionated through 1% agarose-formaldehyde gel and transferred to nylon filters and then hybridized to 32P-labeled DNA probes and placed for autoradiography. Levels of specific RNAs were determined by computer-assisted laser densitometry. Blot filters were sequentially hybridized to nm23 and TIMP-1 probes in addition to GAPDH, as a control. Statistical analyses were carried out according to t-test for equality of means.


RESULTS
NM23-H1 were detected in each sample, however it did not correlate with malignancy and invasiveness. On the other side TIMP-1 gene expression showed a clear correlation between low expression and invasiveness.


CONCLUSIONS
The data suggest that TIMP-1 is an inhibitor of high grade gliomas invasion. NM23-H1 was present in the entire gliomas sample, but it did not vary in diffuse astrocytomas and glioblastomas.
Nasser
José Augusto
JA
Escola Paulista de Medicina, Universidade Federal de São Paulo, SP-Brazil, Universidade Estácio de Sá, Avenida Ataulfo de Paiva 135/914, 22440-032 Rio de Janeiro RJ, Brazil. nasser@riodejaneiro.br
Falavigna
Asdrúbal
A
Ferraz
Fernando
F
Duigou
Gregory
G
Bruce
Jeffrey
J
D001254:Astrocytoma / Q000235:genetics*; D015152:Blotting, Northern; D001932:Brain Neoplasms / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D005819:Genetic Markers; D005909:Glioblastoma / Q000235:genetics*; D006801:Humans; D054778:NM23 Nucleoside Diphosphate Kinases; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000235:genetics*; D009701:Nucleoside-Diphosphate Kinase / Q000235:genetics*; D012334:RNA, Neoplasm / Q000032:analysis; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics*; D014158:Transcription, Genetic
D005819:Genetic Markers; D054778:NM23 Nucleoside Diphosphate Kinases; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm; D019715:Tissue Inhibitor of Metalloproteinase-1; C517938:NME1 protein, human; D009701:Nucleoside-Diphosphate Kinase