The multistep aspects of carcinogenesis including initiation and promotion problems in both human and experimental hepatocarcinogenesis are discussed, especially in terms of oncogene and antioncogene changes. It is shown that the H-ras activation may be an event occurring in relatively late phase of carcinogenesis in the mouse systems, and that hepatitis B integration frequently causes host chromosomal rearrangement possibly leading to inactivation of cancer suppressor genes. In addition, significance of endogenously-produced nitrosamines in hepatocarcinogenesis is pointed out.

A 63 year-old woman was referred to our hospital because of fever and increased number of blasts in the bone marrow. On physical examination she had slight hepatomegaly but no splenomegaly. Laboratory tests disclosed a hemoglobin level of 8.5 g/dl; a WBC count of 13,200/microliter with 26% blasts; a platelet count of 51,000/microliter. A bone marrow aspirate was normocellular with 74% blasts and 37% blasts were stained positive for myeloperoxidase. Cell surface markers for HLA-DR, CD10, CD19, CD13, CD33 were positive. Karyotype analysis revealed 46, XX, t (9q+; 22q-) and 45XX, -7, t (9q+; 22q-). Southern analysis showed rearrangement of immunoglobulin heavy chain but not T cell receptor beta gene. Rearrangements in M-BCR were not detected with 5' or 3' bcr probes. After 2 courses of chemotherapy, blasts decreased to 7% with recovery of normal elements and 11 out of 20 metaphases of the bone marrow cells were normal karyotype. These findings suggest that this case was de novo Ph1 positive acute leukemia which demonstrated both lymphoid and myeloid features.

Bilateral tumors of the same histotype occur rarely in opposite organs. These include hereditary diseases such as retinoblastoma and Wilm's tumor. Bilateral tumors of the same histotype also occur in the parotid glands. A study was performed to determine if there is a possibility that hereditary tumors are included in such cases. Two cases of bilateral parotid tumors, one of pleomorphic adenoma and one of adenolymphoma were reported, and 59 cases reported between 1942 and 1990 with clear times of onset which underwent histological examinations were analyzed. The main histotypes were adenolymphoma in 50.8% (30 cases), pleomorphic adenoma in 27.1% (16 cases), acinic cell tumor in 13.6% (8 cases). Onset of the tumors was simultaneous in 43% of the adenolymphomas. The sex and age at onset showed the same trends as in unilateral cases for each histotype. Among parotid tumors, the percentage of bilateral onset is 1-2%, and when adenolymphomas which show the highest incidence among such tumors were considered from the standpoints of multiplicity, simultaneous onset and the fact that they are benign, the possibility that hereditary diseases are included in the aforementioned 43% was suggested.

We report a 63-year-old woman with renal cell carcinoma associated with von Hippel-Lindau disease. The patient was referred to the department of neurosurgery at our hospital, complaining of gait disturbance. There was a history of retinal hemangioma. After further examination, von Hippel-Lindau disease was the conclusion with evidence of cerebellar hemangioblastoma. An abdominal CT-scan and arteriography revealed multiple hypervascular tumors in the right kidney, but not in the left. She underwent a right radical nephrectomy and lymphadenectomy 2 months after resection of a brain tumor. Von Hippel-Lindau disease is generally recognized as an autosomal-dominant inherited disorder. The patient had a positive family history, seen in both her younger brother and son. Both were diagnosed with renal cell carcinoma with central nervous system involvement before any sign of disease was found in the patient. Twenty one cases of von Hippel-Lindau disease associated with renal tumors have been reported in the Japanese literature. The clinical findings of these cases are discussed.

On the purpose of elucidating the malignant potential, flow cytometric DNA analyses were carried out using paraffin-embedded materials of 82 subserosal, serosal and adventitial invasive colorectal carcinoma. The ratio of DNA diploidy against DNA aneuploidy was one to two. DNA ploidy pattern was not correlated with histological grade but with histopathological factors such as lymphatic permeation, lymph node metastasis and venous invasion. At the primary operation, the rate of the liver metastasis in DNA aneuploid cancer was 16.4% but in DNA diploid cancer the liver metastasis was not observed at all. On the liver metastasis, the 5-year disease free survival rates were lower in patients with DNA aneuploidy (72.8%) than those with DNA diploidy (95.2%). These results indicate that DNA ploidy pattern is related to liver metastasis. Furthermore, the 5-year survival rates in patients with absolute curative resection were lower significantly in DNA aneuploidy (62.5%) than DNA diploidy (92.9%). In conclusion flow cytometric DNA analysis is useful for evaluating the biological malignant potential and predicting the liver metastasis in colorectal carcinoma.

Expression of human epidermal growth factor (EGF) was examined immunohistologically in 93 surgically resected gastric carcinomas, using Biotin-StreptAvidin method pretreated by protease, and its relation with cancer progression and DNA ploidy pattern was studied. DNA ploidy pattern was determined by cytofluorometric measurement. The gastric cancers were divided into two basic ploidy patterns; a diploid mode and an aneuploid mode. EGF expressions were found in 5 out of 20 cases (25%) in intramucosal carcinomas and, the more deeply the carcinomas invaded into the gastric wall, the greater the frequency of EGF expressions did not always become. In the carcinomas with the aneuploid pattern, EGF expression was found only in 2 out of 12 cases (17%) in early cancers and significantly increased in advanced cancers, 15 out of 22 cases (68%). On the other hand, in the carcinomas with the diploid pattern, EGF was expressed in 9 out of 31 cases (29%) in advanced cancers, not significantly different in early cancers, 11 out of 28 cases (39%). Consequently, in advanced cancers, EGF was found more frequently in the aneuploid tumors than in the diploid tumors. These results suggested that EGF expression in the gastric carcinomas is closely correlated with the progression to the advanced cancer with DNA aneuploidy.

Growths factors, defined as polypeptides that stimulate cell proliferation, are major growth-regulatory molecules for cells in culture and probably also for cells in vivo. Evidence has been derived for autocrine system in which the cell produces its own growth factor. Several growth factors as well as their cellular receptors have been identified as productions of proto-oncogenes. Furthermore, these growth factors have been identified as mitogens in tumors of the central nervous system. The roles of growth factors including platelet-derived growth factor (PDGF), epidermal growth factor (EGF) and its receptor. Insulin-like growth factors (IGFs), transforming growth factors (TGFs) and fibroblast growth factor (FGF) on the proliferation of brain tumors, especially glioma were reviewed. The activation of cellular proto-oncogenes resulting in the autocrine system of growth factors and their receptors offers the opportunity for therapeutic interference. Therapeutic efforts will be based on the concepts of neutralization of growth factors, antagonizing growth factors at their receptors, irreversibly blocking receptors, and interference with oncogene product synthesis. Specific antibody for growth factors or receptors will be able to inhibit the proliferation. Trapidil, an antagonist for PDGF, can inhibit the proliferation of a PDGF-producing glioma cell. We can assume that the further analysis of growth regulatory mechanism will allow the design of new therapeutic approaches.

The prediction of prognosis and the design of treatment modalities for patients with brain tumors are judged mostly by its histopathology. The morphology of each tumor, however, represents a part of its phenotype, and more attention should be paid to understand phenotypic expression of individual tumors in relation to cytogenetic abnormality at the molecular level, proliferative potential and invasiveness. The author reviewed current studies on cytogenetics, growth characteristics of various brain tumors, and experimental approaches for invasiveness.

The 9-year-old boy was admitted to Shizuoka Children's Hospital because of cervical lymphoadenopathy. Complete blood count showed normal RBC and platelet counts. WBC was 2700/microliters with no tumor cells. Bone marrow aspirate showed normocellularity with 34% tumor cells. Lymph node biopsy from his right neck was performed and the patient was diagnosed as non-Hodgkin's lymphoma (lymphoblastic type). Surface marker analysis disclosed that the tumor cells were positive for CD5, CD7, CD19, CD38, CD71, and Ia antigen. Chromosomal analysis of the cervical lymph node revealed 46, XY, t(7;14) (p15;q32). Molecular investigation with appropriate probe showed germ-line configurations of IgH gene, TcR beta gene, and TcR gamma gene, and one rearranged band of TcR delta gene. Monoclonality of tumor cells was demonstrated from chromosomal analysis and molecular study. CD7 and CD19 are not lineage specific antigens because CD7 is expressed on immature AML cells and CD19 is expressed on T ALL cells or AML cells. Moreover, TcR delta rearrangement is considered to occur at early phase of hematolymphoid cells. Based on these data, tumor cells of this patient is considered to originate from immature lymphoid cell, so-called lymphoid stem cell.

The expression of proliferation-associated nuclear antigen p105 of gastric carcinomas was studied by multiparameter flow cytometry and immunohistochemical technique. Content of p105-antigen in cancer cells increased with cell cycle progression, and increased more rapidly in cells in late-S phase than in cells in G0 and early S-phases. Cells in M-phase exhibited a dramatic increase in the amount of the antigen, and the amount of IF intensity of mitotic cells were approximately 5-10-fold greater than that of cells in G1- and S-phases. Immunohistochemical technique demonstrated that patients with lymph node metastasis are more likely to have high p105 positive rates than node-negative patients. The mean p105 positive rates of aneuploid tumors were significantly higher than those of diploid tumors. These results indicate that the measurement of p105 positive rates may be a powerful prognostic indicator of gastric carcinoma.

The existence of tumor-suppressor genes has been primarily suggested by three lines of evidences: 1) the suppression of transformed phenotypes of tumor cells by cell-cell hybridization with normal cells; 2) non-random chromosome deletions in a variety of tumors; 3) loss of heterozygosity in specific chromosomal regions in tumor cells. Results from monochromosome transfer experiments also suggest the existence of multiple, functionally distinct tumor-suppressor genes. Recently, several tumor-suppressor genes, which appeared to be functionally distinct, (i.e., Rb gene, WT gene and DCC gene) were isolated. Most recently, it was suggested that the inactivations of at least three different tumor-suppressor genes were required for the colorectal carcinogenesis at different steps. Thus, these findings support that losses or alterations in the dosage of multiple tumor-suppressor genes play crucial roles during initiation and/or progression of a wide variety of cancers.

The nuclear area, N/C ratio and anisonucleosis by morphometry and the DNA content by image photo-cytometry of paraffin-embedded tissues of 9 cases of prostatic hypertrophy and 48 cases of carcinoma obtained by open surgery, needle biopsy or by autopsy were correlated with histological grading. In histological grading, in addition to 9 benign hypertrophy cases, 11 were classified as well differentiated adenocarcinoma, 19 as poorly differentiated adenocarcinoma and 1 as small cell carcinoma. All 14 tumors taken from distant metastasis were classified as poorly differentiated adenocarcinoma. Correlation of the data by tumor cell morphometry with those of nuclear image photo-cytometry was carried out on surgical materials alone. In autopsy materials, DNA ploidy patterns were compared between primary lesions and metastases. The DNA histogram patterns of most cases were classified into 4 groups. Hypertrophy was differentiated from carcinoma by the histogram patterns. There were significant correlations between mean DNA value and morphometric factors. One of the 4 histogram patterns had a higher mortality than others. The DNA histograms did not distinguish between the primary and metastatic lesions of the same patients. However, the DNA histograms constructed from metastasis frequently showed a single stem-line of the primary lesion. This study disclosed several correlations between histological grading and morphometrical factors.

To assess the frequency and significance of 14q32 translocation abnormalities in childhood acute lymphoblastic leukemia (ALL) and the differences between the clinical and cytogenetic features of patients with the 8; 14 translocation and those of patients with other 14q32 translocations, we analyzed our experience with 124 consecutive cases with completely banded karyotype. Eight cases (6.5%) with 14q32 translocation were identified :5 with the 8; 14 translocation and 3 with other 14q32 translocations. As compared with ALL children lacking 14q32 translocations, these 8 cases had a higher serum lactic dehydrogenase (LDH) level, more L3 (FAB classification), and a poorer outcome. On the other hand, in comparison with ALL patients with other 14q32 translocations, patients with the 8:14 translocation were likely to be younger (median age 4.5 years vs 10.4 years), to have a higher serum LDH level (median 5832 IU/l vs 504 IU/l), to have more L3 (3/5 vs 0/3), to have a higher induction failure rate (4/5 vs 1/3), and to have more partial duplication of the long arm of chromosome 1 (4/5 vs 0/3). These results helped clarify the characteristic features of ALL children with 14q32 translocations and showed that ALL children with the 8 ; 14 translocation have different clinical and cytogenetic findings from those of ALL children with other 14q32 translocations.

Molecular mechanism of development and progression of gastric cancer which could be a base of molecular diagnosis was described. Amplification and point mutation of oncogenes are less common in gastric carcinomas, even though it is valuable for diagnosis. Amplification of ERBB2 seems to be an indicator for metastatic ability of gastric carcinoma. Overexpression of EGF/receptor system is a biologic marker for high malignancy. Diagnostic significance for scirrhous gastric carcinomas is found in over-expression of TGF beta, IGF and PDGF. Loss of heterozygosity on chromosomes 5q and 17p frequently occurs commonly in well differentiated type gastric cancer. More accumulation of molecular alterations in the development and progression of gastric cancer should make the molecular diagnosis more valuable in clinical field.

Usefulness of DNA analysis in diagnosis of hematopoietic malignancy was discussed. Examination on the presence of rearrangement in immunoglobulin (Ig) and T cell receptor (TCR) was the first DNA analysis used for clinical diagnosis of lymphoid malignancy to determine the cell-lineage and clonality of proliferating lymphoid cells. One point mutation in ras oncogene has also been used to detect residual leukemic cells as well as diagnosis of the early relapse of leukemia, although not all leukemic cells have this mutation. Presence of BCR-abl fused gene is a genetic marker for Ph1 chromosome. Analysis of BCR-abl gene has made it possible to diagnose the Ph1 ALL and masked Ph1 CML. Development of PCR technique markedly increased the possibility for the use of DNA analysis in clinical medicine. In addition to Ph1 chromosome, various chromosomal abnormalities resulted in a reciprocal translocation between Ig or TCR gene and other genes in various lymphoid malignancies, such as Burkitt lymphoma and follicular lymphoma. These translocations can be analyzed by Southern hybridization and used for clinical diagnosis.

Several methods for DNA analysis including DNA histogram and proto-oncogene amplification in human breast cancer, and the results recently reported were reviewed. A large number of DNA histograms obtained by flow cytometry have provided a possible correlation between DNA ploidy pattern and clinical outcome of breast cancer patients. Poor clinicopathologic factors, however, are not always in association with DNA aneuploidy and/or S-phase fraction rate, suggesting that further investigations on DNA ploidy are required. Amplification and/or over expression of proto-oncogenes in human breast cancers have been reported. Among them c-erbB-2 amplification may be one of the candidates for prognostic indicators of breast cancer survival. To determine any reliable biological factors exist further analysis of tumor DNA will be required in breast cancer research.