Mapping of metastasis suppressor genes for prostate cancer by microcell-mediated chromosome transfer.

AIM: To identify the metastasis suppressor genes for prostate cancer. METHODS: A copy of human chromosomes was introduced into the highly metastatic Dunning R-3327 rat prostate cancer cells by the use of microcell-mediated chromosome transfer. Relationships between the size of human chromosomes introduced into microcell hybrid clones and the number of lung metastases produced by the clones were analyzed to determine which part of human chromosomes contained the metastasis suppressor gene(s) for prostate cancer. To determine portions of human chromosomes introduced, G-banding chromosomal analysis, fluorescence in situ hybridization analysis, and polymerase chain reaction analysis were performed. RESULTS: Each of microcell hybrid clones containing human chromosomes 7, 8, 10, 11, 12, or 17 showed decreased ability to metastasize to the lung without any loss of tumorigenicity. This demonstrates that these human chromosomes contain metastasis suppressor genes for prostate cancer. Spontaneous deletion of portions of human chromosomes was observed in the human chromosome 7, 10, 11, 12, and 17 studies. In the human chromosome 8 study, irradiated microcell-mediated chromosome transfer was performed to enrich chromosomal arm deletions of human chromosome 8. Molecular and cytogenetic analyses of microcell hybrid clones demonstrated that metastasis suppressor genes on human chromosomes were located on 7q21-22, 7q31.2-32, 8p21-12, 10q11-22, 11p13-11.2, 12p11-q13, 12q24-ter, and 17pter-q23. KAI1 and MKK4/SEKI were identified as metastasis suppressor genes from 11p11.2 and 17p12, respectively. CONCLUSION: This assay system is useful to identify metastasis suppressor gene (s) for prostate cancer.

Photographic detection of fluorescent-labelled oligodeoxynucleotide in the blotting format by peroxyoxalate chemiluminescence.

The preparation of a fluorescent labelled oligonucleotide and its photographic detection by peroxyoxalate chemiluminescence (PO-CL) are described. Fluorescent labelling of an oligonucleotide (15-mer) was performed with naphthalene-2,3-dicarboxaldehyde to give an N-substituted 1-cyanobenz[f]isoindole (CBI) derivative (CBI-15-mer). For the photographic detection of CBI-15-mer, the bis(2,6-difluorophenyl) oxalate (DFPO)-dimethyl phthalate (DMP) system was selected to obtain a long-lived CL emission. After optimizing the conditions for the CL reaction, the system was applied to the photographic detection, and as little as 250 fmol per spot of CBI-15-mer on a membrane were detected as a visible spot with an instant photographic film.

[Adaptability of composite resin inlays. 1. Adaptability to the experimental MOD cavity prepared in an acrylic block].

The adaptability of composite resin inlays was examined. CR Inlay and INLAY/ONLAY were used as materials. After the CR Inlay was polymerized in a U-form metal mold, the dimensional changes were measured before and after heating. Furthermore, the composite resin inlays made according to the manufacturer's instructions were cemented in an MOD type cavity and the film thickness of the cement was measured. The polymerization shrinkage of CR Inlay was 0.63% in the inside of the inlay and after heating it increased to 0.71%. The film thickness of the cement in the case of CR Inlay was more than 250 microns and that in the case of the INLAY/ONLAY was more than 750 microns in the MOD type cavity. Dentacolor XS (a box type photo generator) was better than Wite Lite (a handy type photo generator) for the polymerization of CR Inlay. Because of the poor adaptation of the inlays and the difficulty in removing a inlay from a stone model for CR Inlay, perhaps polymerization shrinkage of the inlay should be compensated in some way.