TY - JOUR
T1 - Cloning of differentially expressed genes in highly and low metastatic rat osteosarcomas by a modified cDNA-AFLP method
AU - Fukuda, Tomokazu
AU - Kido, Akira
AU - Kajino, Kazunori
AU - Tsutsumi, Masahiro
AU - Miyauchi, Yoshizumi
AU - Tsujiuchi, Toshifumi
AU - Konishi, Yoichi
AU - Hino, Okio
N1 - Funding Information:
This study was supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science, Sports, and Culture of Japan and the Organization for Pharmaceutical Safety and Research (OPSR) Abbreviations used: PCR, polymerase chain reaction; cDNA-AFLP, cDNA-amplification fragment length polymorphisms; SOS, osteosarcoma-spontaneous origin; S-SLM, selected lung metastasis lesion.
PY - 1999/7/22
Y1 - 1999/7/22
N2 - To identify differentially expressed genes between highly and low metastatic rat transplantable osteosarcomas, we applied a modified AFLP (amplified fragment length polymorphisms) method for cDNA subtraction. The specific point of our modification is selective amplification using suppression PCR technique after restriction enzyme cutting. Our cDNA-AFLP gave high reproducibility (about 95%) in mRNA patterns and enabled us to clone four dominantly expressed genes in a highly metastatic tumor line. Three showed homology with known genes, encoding Ki-67, a proliferation-associated effective marker of malignancy, type IV collagen alpha-3, a major component of basement membrane, and KIAA77 for which the function is unknown. Although one fragment showed no database homology, we revealed a derivation from the rat homologue of the Drosophila melanogaster diaphanous gene (Dia) by cloning of longer cDNA. Dia genes, known to affect actin filament formation, are downstream effectors of Rho small GTPase. The results suggest that alterations in the expression of cytoskeletal protein, basement membrane elements, and proliferative markers may be important for metastasis of osteosarcomas.
AB - To identify differentially expressed genes between highly and low metastatic rat transplantable osteosarcomas, we applied a modified AFLP (amplified fragment length polymorphisms) method for cDNA subtraction. The specific point of our modification is selective amplification using suppression PCR technique after restriction enzyme cutting. Our cDNA-AFLP gave high reproducibility (about 95%) in mRNA patterns and enabled us to clone four dominantly expressed genes in a highly metastatic tumor line. Three showed homology with known genes, encoding Ki-67, a proliferation-associated effective marker of malignancy, type IV collagen alpha-3, a major component of basement membrane, and KIAA77 for which the function is unknown. Although one fragment showed no database homology, we revealed a derivation from the rat homologue of the Drosophila melanogaster diaphanous gene (Dia) by cloning of longer cDNA. Dia genes, known to affect actin filament formation, are downstream effectors of Rho small GTPase. The results suggest that alterations in the expression of cytoskeletal protein, basement membrane elements, and proliferative markers may be important for metastasis of osteosarcomas.
KW - Metastasis
KW - Osteosarcomas
KW - Subtraction
KW - cDNA-AFLP
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U2 - 10.1006/bbrc.1999.0997
DO - 10.1006/bbrc.1999.0997
M3 - Article
C2 - 10405319
AN - SCOPUS:0033595146
VL - 261
SP - 35
EP - 40
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -