A 65-year-old male was admitted to our hospital due to recurred malignant lymphoma of left tonsil origin. Studies using flow cytometry on mononuclear cells in peripheral blood revealed the appearance of intermediate B cells, and examinations on gastrointestinal tract showed diffuse infiltration of medium-sized lymphoid cells into stomach and colon, that had the same phenotype as tumor cells in peripheral blood. They suggested leukemic change and gastrointestinal tract infiltration of malignant lymphoma. Southern blot analysis revealed T-cell receptor beta-chain gene rearrangement as well as immunoglobulin gene rearrangement. Analysis by histo in situ hybridization on infiltrated gastric mucosa specimen showed diffuse expression of immunoglobulin mRNA in tumor cells, but no message of T-cell receptor beta-chain gene. Northern blot analysis showed the same result. It suggests that in this case, the rearrangement of T-cell receptor beta-chain gene is ineffective rearrangement without transcription to mRNA.

Clonal evolution which characterizes malignant tumors is the consequence of two antagonizing forces acting on the tumor cell population, namely, forces of diversion and conversion. The former makes cells to diverge through genetic and epigenetic instabilities which are the built-in characteristics of malignant cells. Possible causes of genetic instability are discussed. These include mistakes in DNA synthesis by an error-prone DNA polymerase, the nucleotide pool distartion and the overreplication of replication origins, abnormal DNA repair, high rate recombination, by expression of fragile sites and possibly by expression of retrotransposons, frequent nondisjunction of chromosomes as a consequence of gene dosage inbalance, and abnormal DNA methylation. The second force makes the resulting tumor cell population with heterogenous phenotypes to converge through selection by host defence mechanisms, competition for nutrients and oxygen among tumor cells, to cell interactions within tumor and between surrounding normal tissues. Genetic tagging of tumor cells with pSV 2neo facilitates the analysis of clonal evolution which results from diversion and conversion of tumor cells. Selective growth and metastasis of a clone in a mouse sarcoma population was demonstrated. Generation of dominant clones as well as drug resistant clones in tumor can be studied with this method.

Based on the two mutation hypothesis in the development of retinoblastoma, loss of heterozygosity (LOH) of specific chromosome has been implicated in the presence of tumor suppressor gene. Studies on the LOH in different types of tumors revealed that LOH of each chromosome might play a different role in the multistep process of carcinogenesis: LOH of some chromosomes may play an etiological role in the development of some tumors, while that of other chromosomes or the same chromosome in other tumors, may play a role in the progression of tumors. LOH of chromosome 13 is an example for the former cases, and the latter cases involve LOH of chromosome 17 in colorectal carcinoma and osteosarcoma, chromosome 10 in glioblastoma, chromosome 1 in neuroblastoma and malignant melanoma, and chromosome 11 in breast carcinoma. These studies indicates that the progressive or concerted LOH could be a measure of the highly malignant or metastatic potentiality. However, it should be borne in mind that, especially in polyploid tumors, LOH also occurs as a random event following the polyploidization-segregation process.

Two non-tumorigenic variant cells were isolated from UV-irradiated BALB/c 3T3 cells according to their morphological responsiveness to phorbol ester tumor promoters. They exhibited remarkably different metastatic behavior after intravenous injections of their v-src transformants into nude mice; phorbol ester-resistant variant TR 4 cells transformed by v-src were hyper-metastatic, whereas v-src transformants of phorbol ester sensitive variant TR 5 cells were not metastatic at all. No different metastatic responses were observed with v-k-ras induced transformants of the variants. These non-tumorigenic variant cells may pre-acquire the genetic alteration of certain src-specific specific and metastasis-associated factors. This system may be useful for genetic analysis of the induction of metastasis.

Alterations in the structure and/or expression of oncogenes have been examined to comprehend better the relationship between metastasis and oncogenes. There are reports that amplification and overexpression of N-myc (human neuroblastoma) and erbB-2 (human breast cancer) are closely related to malignant progression. On the other hand, DNA transfer experiments have also been done to assess involvement of oncogenes in metastasis. The active ras oncogenes, which are frequently used, show that transfer of the ras oncogenes endow and/or enhance the metastatic potential of the recipient cells. In addition to the phenomenological studies to determine whether an oncogene alters metastatic potential, increasing attention has been directed to possible phenotypic changes relating to metastasis and affected by oncogenes. It has been clarified that transfer of a certain oncogene, such as ras and fos, alters the expression of biomolecules, for example, metalloproteinases responsible for invasion, autocrine motility factors and cytoskeletal proteins related to cell motility, receptors of fibronectin for cell attachment, and histocompatibility antigens, such as H-2K and H2-D. While further characterization of the biomolecular changes induced by the oncogenes has to be done, the mechanisms of alteration in the expression and function of biomolecules directly involved in the metastatic potential also should be paid attention.

In order to understand the role of myc family genes (c-myc, N-myc and L-myc) in the development and progression of human testicular cancer, we have analyzed the expression of myc family genes in three different types of primary human testicular cancer (seminoma, embryonal carcinoma and teratocarcinoma) and normal testis using Northern blot analysis. Expression of N-myc gene, which is usually limited in the neoplasms derived from neuroectoderm, was detected in seven out of ten cases of seminomas and two out of two cases of embryonal carcinomas. Gene amplification was not observed in these cases. Expression of N-myc gene was not detected in teratocarcinomas and normal testes. Expression of c-myc gene was observed in seminomas, embryonal carcinomas, teratocarcinomas and normal testes, but specific expression of c-myc gene was not seen in these cancers and normal testes. Expression of L-myc gene was not detected in all cases examined in our studies. Since N-myc gene expression was observed only in undifferentiated testicular neoplasm, such as seminoma and embryonal carcinoma, its expression may be positively related to the development and progression of special types of human testicular cancer.

A 12-year-old boy was referred to our hospital because of anemia and jaundice. On admission bone marrow smears were compatible with M6 classification of the FAB, revealing 74.5% erythroblasts of all nucleated cells and 40% blasts of nonerythroid cells. Karyotype analysis revealed 46, XY. Gene rearrangement within the breakpoint cluster region (bcr) on chromosome 22 was negative at this time. Complete remission was attained by a combination chemotherapy. However, at 10 months of remission, cytogenetic studies of the bone marrow demonstrated Ph1 positive (10%). One month later, the patient fully relapsed with a 75% Ph1 positive karyotype associated with positive bcr. Subsequently, the patient died of refractory leukemia.

A 71-year-old woman was hospitalized because of hematemesis on December 1, 1987. Her white blood cell (WBC) count was 41,200/microliters with 48% of lymphoblasts, and the bone marrow was hypercellular with more than 90% of blasts. The diagnosis of acute lymphoblastic leukemia (ALL) (FAB: L3) was made by morphologic, cytochemical and immunologic studies of the blasts. The examination of fiber gastroscope revealed remarkable varix in the stomach, suggesting portal hypertension accompanied by infiltration of leukemic cells into reticulo-endothelial system. She died of respiratory failure because of bleeding into the trachea. The autopsy disclosed the massive infiltration of leukemic cells into the whole organs. In the chromosome study of the peripheral blood, t(2; 8) and 14q+ were observed, and these chromosomal abnormalities are relatively unusual in patients with Burkitt's lymphoma.

The chromosome 14q11 anomaly has been reported to be specific to adult T-cell leukemia (ATL) and this anomaly has also been confirmed in preleukemic state of ATL (pre-ATL) patients though the frequency is low. In an attempt to clarify if the same chromosome aberrations could be found also at the stage of HTLV-I carrier and if there is any cytogenetic difference from non-HTLV-I carriers, a cytogenetic study of lymphocytes stimulated with phytohemagglutinin in three HTLV-I healthy carriers and three non-HTLV-I carriers in an ATL family was performed. The results were as follows. 1. In three HTLV-I carriers, 7 of 311 cells examined (2.3%) showed chromosome aberrations, and 4 cells (1.3%) had 14q11 anomaly. 2. In three non-HTLV-I carriers, 4 of 260 cells examined (1.5%) showed chromosome aberrations, whereas no cells had 14q11 anomaly. These findings suggest that 14q11 anomaly is already present at the stage of HTLV-I carrier and seems to be an important cytogenetic clue to the pathogenesis of ATL.

To clarify the relationship between oncogene c-sis expression and tumorigenesis in human osteosarcoma, in situ hybridization and immunohistochemical studies were performed to detect c-sis mRNA and PDGF-like protein partially consisting of c-sis product. Formalin fixed-paraffin embedded sections of eight cases of osteosarcoma of bone were examined. Three osteosarcomas highly expressed c-sis mRNA with a fine granular pattern in their cytoplasms. Five osteosarcomas, including three cases with c-sis expression, contained PDGF-like protein in their cytoplasms. These results suggested that c-sis oncogene and PDGF-like protein were closely related to tumorigenesis in human osteosarcoma. The DNA-mRNA in situ hybridization technique applied by the author is as efficient as the immunohistochemical method in cancer research.

This include review of recent paper about sensitivity and effective of radiation therapy using flow cytometry. This paper discuss the prediction of tumor response. As predictor of response in cancer. It is DNA aneuploidy, growth fraction (% S fraction), chromosome changes, and monoclonal antibody by oncogene product protein. It is possible that the protocol of cancer treatment will be determined based on prediction data from an individual cancer. Flow cytometry would be a useful tool to get a predictor of radiation, drugs response on each cancer.

A solitary hemangioblastoma of the optic nerve was found in a 36-year-old male with a distinct family history of intracranial hemangioblastoma. The patient was admitted with complaints of visual loss and exophthalmos of the right eye. X-rays showed enlargement of the right optic canal. Right carotid angiography revealed a hypervascular tumor in the orbital apex, supplied by the ophthalmic artery. Computed tomography disclosed a pear-shaped, isodense mass with moderate contrast enhancement in the orbital apex. The right optic nerve was enlarged along its entire course and was involved with the apical mass. Surgery via the right frontal extradural approach disclosed a solid, vascular tumor involving the optic nerve at the apex of the orbit. Histological examination showed the tumor to be a characteristic hemangioblastoma. Over 90% of intracranial hemangioblastomas are located in the posterior fossa. Supratentorial hemangioblastomas, especially those arising in the optic nerve, are extremely rare. In addition, optic nerve hemangioblastomas are frequently familial and are associated with infratentorial hemangioblastomas, angiomatosis retinae, and cysts of the abdominal viscera.

This is a rare case of a 57-year-old-male patient with familial, benign and multiple pheochromocytoma recurring 23 years after initial surgery. His 131I-MIBG scintigraphy was false negative, but his son's 131I-MIBG scintigraphy was true positive.

In the present study, monoclonal antibody rp-28 directed against the ras gene product p21 was used to evaluate ras p21 expression in malignant and benign ovarian tissues. Some ovarian carcinomas (serous cystadenocarcinoma, mucinous cystadenocarcinoma, undifferentiated carcinoma and clear cell carcinoma) demonstrated intense staining of ras p21. In mucinous tumors, both the frequency of ras p21 positive staining and the staining intensity gradually increased with the degree of malignancy. There was no difference in ras p21 expression according to the clinical stages of ovarian carcinomas. In the metastatic lesions, ras p21 staining was rather weaker than in the primary lesions. It is therefore possible that intense staining of ras p21 is associated with the degree of malignancy in some types of ovarian tumors, and that the expression of ras oncogene product p21 is not enhanced with progression and metastasis in such types of ovarian carcinoma. These results suggest that ras oncogene plays an important role in the carcinogenesis of some types of epithelial ovarian tumors.

For the purpose of elucidating the role of oncogenes like c-myc in renal cell carcinoma, the methods of introducing exogenous genes into human cells might be powerful tools. In the present study, the electroporation gene transfer method was investigated for its application to renal cancer cells. A mixture of ACHN cells (human renal cancer cell line, which cannot grow in the presence of neomycin) and DNA of neomycin-resistant genes with SV 40 promoters was exposed to electric pulses from an electroporated (Bio-Pulser 101, UNISOKU). And the cells were cultured in a medium containing Geneticin (neomycin analog) for 3 weeks. Then, the number of formed neomycin-resistant neoR colonies was counted. In the cells of neoR colonies, the existence of neoR genes and their expression were confirmed by Southern and Northern blot gene analyses. The transformation efficiency (number of neoR colonies/inoculated cells) was positively correlated with cell densities, DNA concentrations, and discharged voltages under our experimental conditions. The transformation efficiency was 1.3 - 3.3 x 10(-4)/cell in the condition of 1 x 10(7) cells/ml, 1 microgram DNA/10(6) cells, and 2 kV/cm. These results suggest that the electroporation gene transfer method is applicable for the study of phenotypic alterations after introducing oncogenes into human renal cancer cells.