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A recent integrative analysis using a phosphoproteomic approach identified FAM129B, also known as MINERVA, as a downstream effector of the MAP kinase pathway in human melanoma cells. FAM129B protein, which is a member of a small family of proteins, was also found to suppress TNFα/cycloheximide-induced apoptosis in HeLa cells. To investigate the physiological functions of Fam129b in vivo, we generated gene-targeted Fam129b-mutant mice in which, the amino terminal coding exon was replaced by lacZ. We found that homozygous mutant mice are viable and fertile and that Fam129b is considerably expressed in most of the epidermal keratinocytes of both embryonic and adult mice. Although no gross defect was observed in the skin of the Fam129b-deficient mice, wound healing subsequent to skin puncturing was significantly delayed. Furthermore, overexpression of Fam129b promoted HaCaT cell motility in an N-terminal pleckstrin homology domain-dependent manner, but not proliferation. Microarray analysis of the Fam129b transfectant exhibited substantial upregulation of several genes related to wound repair and cell motility. These results suggest that expression of Fam129b in epidermal keratinocytes accompanied by alteration of wound healing-related gene expression is necessary for regulation of cell motility and thereby, contributes to the appropriate wound healing process.

Background: a-Fetoprotein (AFP) and prostate-specific antigen (PSA) in serum are widely used as tumor markers in the evaluation of prognosis and management of patients with hepatocellular carcinoma and prostate cancer, respectively. To establish the molecular diagnosis of cancer, reverse transcriptase polymerase chain reaction (RT-PCR) for AFPand PSA was used to identify circulating cancer cells in the blood of cancer patients. Here, we examined the tissue-specificity of AFPand PSA and tested whether AFPand PSA are suitable targets in the detection of certain cancer cells by RT-PCR using peripheral blood samples. Methods: Tissue specificity of AFP and PSA was analyzed by Northern blotting and RT-PCR. Probes for AFPand PSA were hybridized with poly A+ RNAs from 50 human tissues. RT-PCR for AFP and PSA mRNA was performed using several cancerous tissues and normal tissues and peripheral blood cells from seven healthy volunteers. Results: Broad expression of AFP was observed in several tissues and a large amount of AFP mRNA was found in fetal liver. PSA was expressed in prostate, salivary gland, pancreas and uterus. By RT-PCR, AFP and PSA mRNA were detected in several tumors, including salivary pleomorphic adenoma, hilar bile duct carcinoma, pancreatic carcinoma, transitional cell carcinoma of urinary bladder and thyroid papillary carcinoma. Furthermore, AFPand PSA mRNAs were frequently detected by RT-PCR, even in peripheral blood cells from healthy volunteers. Conclusions: Neither AFPnor PSA showed tissue-specific expression. AFPand PSA mRNA were detected in several diseased and non-diseased tissues and normal circulating blood by RT-PCR.

The heterotrimeric guanine-nucleotide-binding proteins (G proteins) and G protein-coupled hormone receptors including gonadotropin receptors have been suggested to play a role in ovarian tumorigenesis. However, no functional significance of gonadotropin receptors and G proteins in this process has been demonstrated. To investigate this issue, we examined point mutations in these genes in various types of ovarian tumors by polymerase chain reaction-single strand conformation polymorphism analysis and direct sequencing. Among 37 tumors (20 epitherial, 8 sex cord-stromal, and 9 germ cell tumors) and 5 carcinoma cell lines examined, no mutational sequence of G protein-interaction domains of luteinizing hormone receptor and follicle-stimulating hormone receptor, or "hot spots" of the alpha subunit of adenylyl cyclase-stimulating G protein and -inhibitory G protein was observed. Although this analysis was performed on only a limited number of tumors and cell lines, and on limited gene loci, the results suggest that mutational activation in gonadotropin receptors and G proteins is not crucial for ovarian tumorigenesis.

Human T-cell leukemia virus type I (HTLV-I) antibody detection has been widely used to screen HTLV-I carriers. Sometimes, however, it gives false positive or negative results. A demonstration of the HTLV-I provirus from patients' peripheral blood mononuclear cells (PBMC) should, therefore, give the crucial evidence for them being HTLV-I carriers. We established a simple and reliable method using the polymerase chain reaction (PCR) to detect one molecule of HTLV-I provirus in 100 x 10(3) PBMC, during which internal control primers for the human beta-globin gene were also employed in the same reaction tube to check the success of the amplification reaction. We can thus easily avoid any false negative judgement and quantitate the HTLV-I provirus in PBMC simply by diluting the sample before PCR. One ml blood was enough for ten or more determinations by PCR. Analysis of seropositive blood from donors demonstrated a wide range for the number of HTLV-I provirus in PBMC. The method could conveniently be used for quantitating HTLV-I proviruses and following up HTLV-I carriers to study the pathophysiology and mode of HTLV-I transmission.