A 73-year-old male was admitted to our hospital in October 1987 because of severe anemia, anorexia, and loss of weight. The hemoglobin level was 5.7 g/dl, the white blood cell count 2,500/microliters with 5% myeloblasts positive for peroxidase, and the platelet count 8.6 x 10(4)/microliters. The LDH was 656 mU/ml, the total protein in the serum 7.4 g/dl, IgG 419 mg/dl, IgA 104 mg/dl, IgM 10 mg/dl, and urine Bence Jones (BJ) protein 8.8 g/day. The X-ray survey of the bones showed multiple osteolytic lesions. A bone marrow aspirate was hypercellular with 91.4% plasma cells, and was cultured a whole day for chromosome study. It revealed an abnormal karyotype of 46, XY, -15, t(6; 14) (p21.1; q32.3), +der(15)t(1; 15) (q23; q24). Immunoelectrophoresis demonstrated lambda type BJ protein. He was treated with melphalan and prednisolone. Proteinuria and marrow plasma cells decreased in amount. In December a white cell count was 6,030/microliters with 80% myeloblasts. A bone marrow aspirate revealed an increase of 82.6% myeloblasts or promyelocytes. The patient was refractory to chemotherapy and died of sepsis in April 1988. An unrelated abnormal karyotype; 48, XY, +8, +13 appeared concomitant with an increase of the leukemic cells, but no cells showed the t(6; 14). We cytogenetically discussed the simultaneous presence of multiple myeloma with acute myelogenous leukemia.

A 46-year-old man was diagnosed as having chronic myelogenous leukemia (CML) in chronic phase in Dec. 1985. In Dec. 1987, anemia and leukocytopenia progressed, and the percentage of blast cells increased in the bone marrow. The blast cells were lymphoblastoid and positive for TdT. It was treated as a lymphoid crisis with vincristine and prednisolone, and complete remission was achieved. However, the blasts (11%) were observed in the bone marrow in Mar. 1988, and the chromosomal analysis revealed 46, XY, t (2q-; 11q+), t (9q+; 22q-) in 13 out of 20 cells. In June, the percentage of the blasts increased again, but chromosomal analysis showed a different karyotype, 46, XY, t(2p-; 11p+), t(9q+; 22q-) which was observed in 9 out of 10 cells. Then, myeloblastoid cells increased rapidly in spite of the chemotherapy in Dec. 1988. The chromosomal analysis showed 46, XY, 2p-, 7q-, 9q+, 11p+, 22q- in all analyzed cells. The rearrangement of the bcr gene could be detected by the Southern blotting. The blasts were positive for CD7, CD11, CD13, CD33, CD36, CD41 and CD42, suggesting that the blasts had the surface phenotypes of both myeloid and megakaryocytoid-lineage. This is a case with the mixed blast crisis that changed from the lymphoid to the myelo-megakaryocytoid in nature, in which three clonal evolutions were observed during the clinical course.

A case of Philadelphia (Ph1) chromosome positive acute myelogeneous leukemia (AML) following a refractory anemia with excess of blasts (RAEB) with 8 trisomy is reported. The 80-year-old man developed pancytopenia during the course of follow-up after the surgical operation of the carcinoma of the sigmoid colon and the rectum for which no irradiation therapy nor chemotherapy had been applied. The diagnosis of RAEB was made according to the diagnostic criteria proposed by FAB co-operative group. Chromosomal analysis revealed 8 trisomy in 54% of the metaphases of bone marrow cells. The remainders showed normal karyotype without Ph1 chromosome. He was on androgenic steroid and activated Vitamin D3 without significant changes in the clinical and the hematological features until 3 months later when many atypical blasts appeared in the peripheral blood. The diagnosis of AML (M2) was made. Chromosomal analysis revealed Ph1 chromosome with the typical 9;22 translocation in 100% of the examined cells. 8 trisomy was not detected any more. Southern blot analysis using bcr probe showed bcr rearrangement. He was treated with a small doses of Ara-C. There was some reduction in the number of blasts in the peripheral blood. However, he died of septicemia 2 months later. The present case indicates that Ph1 positive acute leukemia with bcr rearrangement is not necessarily considered as a blastic transformation of chronic myelogeneous leukemia and such a cytogenic abnormality can appear in a leukemic transformation of myelodysplastic syndrome.

Thirty-four children, including nine relapsed cases with acute lymphoblastic leukemia (ALL) having hyperdiploidy (greater than 50 chromosomes) were studied on clinical and cytogenetic characteristics. The majority of children initially with hyperdiploidy (greater than 50 chromosomes), who showed favorable prognostic features such as lower leukocyte counts, lower serum lactic dehydrogenase levels, ages between 2 and 10 years, or the presence of common ALL antigen, had the most favorable outcome among childhood ALL (5-year survival rate was 100%). Even nine children, who showed poor prognostic features such as ages over 10 years, leukocyte counts over 2 X 10(4)/mm3 or lymphomatous signs, had also the same favorable outcome. There were no differences in clinical features between 6 patients with additional chromosomal structural abnormalities and 19 patients without them. Duplication of the long arm of chromosome 1 was frequently observed as additional chromosomal structural abnormalities. Patients with hyperdiploidy (greater than 50 chromosomes) observed at relapse, who had the same favorable clinical features as those at diagnosis, had a poorer prognosis. These findings show that initial hyperdiploidy (greater than 50 chromosomes) is an independent favorable prognostic sign in childhood ALL and additional chromosomal structural abnormalities may not indicate a poor prognosis among childhood ALL with hyperdiploidy (greater than 50 chromosomes). On the other hand, relapsed children with hyperdiploidy (greater than 50 chromosomes) have not a favorable outcome after the onset of relapse.

Amplifications of myc oncogenes (c-myc, N-myc) were studied by Southern blot hybridization methods in ovarian cancers. Fourteen cases were primary ovarian cancers and one case was metastatic ovarian cancer. Primary and metastatic foci of three primary ovarian cancers were compared. A serous adenocarcinoma had a c-myc gene amplification more than 10 fold without N-myc gene amplification. This case also had an amplification in other c-myc probes with upper and down streams of the second exon. The results suggested that this case had c-myc gene expression. One of these cases had 5 fold amplification of the metastatic focus compared with the primary focus. The amplified c-myc gene did not show relations among clinical courses, prognosis and histologies.

We analyzed the characterization of DNA synthesized cells (S phase cells) in the proliferation. It is important that the fixation and cell cycle time of S phase cells and DNA qualitative changes, etc., be examined. We used the dual-laser FCM system of the Los Alamos National Laboratory in New Mexico and analyzed the cell cycle, especially changes in S phase cells both quantitatively and qualitatively. By the bromodeoxyuridine (BrdU)/Hoechst Quenching effect, we could detect differences in electric signals by adopting the differential fluorescence correction method. (1) The effects of sodium butyrate in gene expression were dose-dependent at human endometrial adenocarcinoma cells. (2) After double staining with Mithramycin and Hoechst33342, S phase cells incorporating BrdU under quenching gave from linear relative to arched bivariate contour histograms. (3) At 24 hours G1 phase synchronization of SB 1,3 and 5mM appeared in slow cycling cells after BrdU was added. These results suggest that the effect of sodium butyrate in human endometrial adenocarcinoma cells might be understood not only from the quantitative change, but also by the qualitative change in S phase cells.

One family of familial gingival fibromatosis was presented. Case 1 is an 11 years old girl. Gingival fibromatosis was noted by delayed eruption of permanent teeth. Fibromatosis was seen in whole gingiva and lower 1/2 to 1/3 of the tooth crowns was covered. Gingivectomy was performed. Case 2 was 42 years old man, who was father of case 1. Firstly gingival swelling was noted at the age of 10 years. He had operations of gingivectomy at the age of 28 years. It recurred and fibromatosis was seen in whole gingiva and lower 1/2 to 1/3 of the tooth crowns was covered. Cytogenetically no abnormality was seen in the chromosomes of both cases. Reported cases of familial and non-familial gingival fibromatosis in Japan were reviewed.

Ras genes are converted to active oncogenes by point mutations occurring in either codon 12, 13 or 61. We used polymerase chain reaction and direct sequence method for the analysis of these mutations. We examined 13 hepatocellular carcinomas, 8 cholangiocarcinomas, 2 hepatoblastomas and 1 biliary cystadenocarcinoma. Of these tumors, ras gene mutations were detected solely in cholangiocarcinomas. Cholangiocarcinoma showed a high incidence of K-ras gene mutation. Among 8 patients with cholangiocarcinoma, the mutation was detected at codon 12 in 4 and at codon 61 in 1. The incidence of K-ras gene mutation was especially high in the hilar type of cholangiocarcinoma as compared with the peripheral type.

A new cell line designated RTSG established in vitro from the pleural effusion of a patient with metastatic ovarian epithelial cancer has been subcultured 46 times for more than 2 years. The cells grew in a monolayered sheet, showing a tendency to pile up, with the population doubling in 48 hrs. Electron-microscopically, desmosomes were characteristically observed, suggesting the cells were of epithelial origin. Chromosomal analysis revealed aneuploidy with a tetraploid mode. The heterotransplanted tumors in nude mice were histopathologically classified as a poorly differentiated adenocarcinoma, whereas the original tumor consisted mainly of mucinous and serous cystadenocarcinoma and only partly of poorly differentiated adenocarcinoma. The cells secreted hCG (38.8 mIU/day/10(6) cells) and beta-hCG (6.1 ng/day/10(6) cells) in spent medium. Immunocytologic +-and-histochemical staining for tumor markers of the original tumor, the cultured cells and the transplanted tumors also revealed the localization of not only hCG and beta-hCG but also CA19-9 and CA-125 whose values had been elevated in the preoperative serum (hCG: 10 mIU/ml, CA19-9: 6,400 U/ml, CA-125: 225 U/ml). Results of PAS, Alcian-blue and Mucicarmine strains indicated that most of the PAS-positive substances in the cultured cells and the transplanted tumors were consistent with glycogen while the original tumor mainly contained mucin except for the lesion of poorly differentiated adenocarcinoma with glycogen. These results suggested that the cultured cells might originate from poorly differentiated adenocarcinoma cells in the original tumor.

A monoclonal antibody, MRK16, recognizing specifically an epitope of P-glycoprotein (P-GP) was used to determine the degree of expression of P-GP in kidney and urinary bladder cancers. Immunohistochemistry, immunoelectronmicroscopy, and immunoprecipitation were used for this study. Expression of P-GP was found in 6 of 20 kidney cancers treated without anticancer drugs. Also expression of P-GP was found in 17 of 47 urinary bladder cancers. 11 of 31 in primary cases, 0 of 5 in recurrent cases treated without anticancer drugs, and 6 of 11 in recurrent cases treated with anticancer drugs were positively expressed. These results indicate that a certain proportion of kidney and urinary bladder cancers intrinsically acquired multidrug resistance, and also that prior administration of anticancer drugs may induce P-GP in initially negative tumors. We also succeeded to detect MDR1 mRNA by means of in situ hybridization. From our present data, our methods to detect P-GP and MDR1 mRNA appeared to be very useful from the point of clinical application.

The C-erbB-2 gene was first found in human genomic DNA as a sequence which had homology in nucleotide sequence to the V-erbB by molecular hybridization under relaxed conditions. The product of this gene is a receptor type protein-tyrosine kinase which has a structure highly related to that of epidermal growth factor receptor (EGF-r: C-erbB-1). The proto-neu gene is a rat counterpart of the C-erb B-2 gene. The C-erbB-2 gene is also called as the HER-2 gene. The C-erbB-2 gene acquires the ability to transform NIH 3 T 3 cells by, 1) mutation which alters valine 659 in transmembrane region to glutamic acid as was found in neu gene activation, 2) deletion of c-terminal regulatory domain or 3) gene-amplification or overexpression. C-erbB-2 expresses in human embryos on mucous membranes and glands, but only faintly in adult tissues. High expression or gene amplification in human tumor appeared to be an indication for high risk of metastasis or high degree of malignancy.

A cell line, HuH-33 was cultured in vitro from a patient with hepatocellular carcinoma. This cell line has been in continuous culture over 12 month period with slow growth potential. HuH-33 was composed spindle-or polygonal-shaped cells as a major population. Chromosome number of the cells were widely distributed even in the primary culture. HuH-33 was transplantable into nude mice and secreted alpha-fetoprotein, albumin, beta 2 microglobulin, ferritin and tissue polypeptide antigen.

The expression of nine proto-oncogenes (c-myc, N-myc, c-fos, c-jun, p53, H-ras, N-ras, c-raf, hst) and other three genes (AFP, PCNA, GST-P) were investigated during spontaneous development to hepatocellular carcinomas (HCCs) in LEC rats. Expressions of c-myc, H-ras, N-ras, c-raf, p53, and PCNA genes were detected but did not significantly change during the development to HCCs in LEC rats. Expressions of N-myc, hst, and AFP genes were not detectable since 5 weeks after birth. Expression of c-fos gene was detected in one out of four HCCs. Significantly increased expression of c-jun gene was observed in the liver tissues of LEC rats aged 8 months. The high expression was decreased in HCCs. On the other hand, the expression of GST-P gene increased in parallel with the clinical course of the development to HCCs in LEC rats. The increased expression of GST-P gene was observed in the liver tissues of LEC rats aged 8 months, and HCCs showed very high expression of GST-P gene. These observations suggest that both c-jun and GST-P genes may play a role in the spontaneous development to HCCs in LEC rats.

Thirty-four primary lung cancers were analyzed for abnormalities in the myc family genes (c-myc, N-myc, L-myc), using the Southern blot hybridization method. They were subcutaneously transplanted into nude mice. The Southern blot analysis showed that c-myc and L-myc genes were amplified in 4 non-small cell carcinomas and 3 small cell carcinomas respectively. Allelic deletion of the L-myc gene was observed in 7 cancers, including 2 carcinomas which also had an additional band of the c-myc gene or amplification of the L-myc gene. No abnormalities in the N-myc gene were observed in this study. Of 13 cancers with abnormalities in the myc family genes, 11 including all tumors with myc gene amplification were transplantable to nude mice. Of 21 tumors without any abnormalities in the myc family genes, however, only 6 were transplantable to nude mice (p less than 0.005). These results indicate that abnormalities in the myc family genes, especially gene amplification might promote tumor-forming capacity in xenotransplantation of lung cancers and this phenomenon might be closely related to the function of the myc gene.

The relationships between the history of cancer in parents and the dietary pattern, serum cholesterol, serum protein and blood hemoglobin were examined to assess the possible role of lifestyle transmission in the familial aggregation of cancer. Individuals with history of cancer in parents were selected from a population of 1,221 individuals. Five age- and sex-matched controls for each case were selected from individuals, the parents of whom were alive at the age of the case parent. Intake of miso soup, pickles and milk product showed associations with the history of cancer in parents. It was also found that serum cholesterol was significantly lower in individuals with history of cancer other than stomach in the mother. Interestingly, serum protein, especially the globulin fraction, was found to be significantly lower among individuals with the history of cancer in parents. These findings suggest that lifestyle transmission might play some role in the familial aggregation of cancer.

Lymphatic permeation (ly) is considered an important prognostic factor, even in the case of a diffuse infiltrative carcinoma (DICA) of the stomach, which generally has the poorest prognosis among the various types of gastric carcinomas. Our previous study has revealed that the ly in DICA is strongly related to the histological type of carcinoma that infiltrates the tumor tissue: the "mixed type", i.e., a type that includes tubular/trabeculoacinar element in 10% or more of the tumor tissue presents a significantly higher incidence of ly than the "pure type", i.e., a type that consists almost exclusively of anaplastic/mucocellular carcinoma. However, even in the "pure type" cases, some show a high ly incidence. To clarify this discrepancy, the DNA ploidy pattern in the "pure type" DICA was investigated, since the DNA ploidy has been reported to correlate well with the invasiveness or the metastatic tendency of the tumor. Although most of the cases (70%) showed a heteroploidy, such an incidence was significantly higher in the cases with a marked ly (86%) than in the cases without ly. (60%). As a result, we have concluded that in cases of a DICA, the lymphatic permeation (ly) is remarkably influenced not only by the histological type seen but also by the DNA ploidy pattern of the carcinoma.