In hybrids SBW I and SBW II between S194 mouse plasmacytoma and BW5147 mouse T-cell lymphoma, enhanced expression of the rearranged c-myc mRNA of heterogeneous sizes (1.8-2.4 kb) in S194 was markedly downregulated but expression of the non-rearranged c-myc of 2.4 kb in BW5147 was not. Treatment of SBW I hybrid cells with cycloheximide augmented non-rearranged c-myc expression 2-4-fold but did not release the downregulated expression of the rearranged c-myc. Nuclear run-on assay showed that the high level of c-myc transcripts in S194 cells declined in SBW I cells comparable to the level in BW5147 cells. The results suggest that the downregulation of the rearranged c-myc in SBW I hybrid cells was at a transcriptional level rather than a posttranscriptional level. Methylation patterns of the rearranged c-myc in SBW I cells was the same as S194 cells. Treatment of the SBW I cells with 5-azacytidine, TPA or forskolin did not release the downregulated expression of the rearranged c-myc, suggesting no causative involvement of DNA methylation or protein phosphorylation in the downregulation. Higher DNase I sensitivity of the rearranged c-myc in S194 cells decreased to a similar extent to that of the non-rearranged c-myc after cell fusion with BW5147 cells. All these results suggest that expression of the rearranged c-myc was downregulated at the level of transcription in murine cell hybrids between a plasmacytoma and a T-cell lymphoma probably by changing the chromatin configuration around the gene from the open to the closed state.

A 3-year-old boy was referred to our hospital in September 1985, because of pancytopenia. His bone marrow was normocellular with 18% blasts, which had Auer rod and were positive for peroxidase staining. A diagnosis of refractory anemia with excess blasts in transformation was made according to FAB criteria. Chromosome analysis of bone marrow cells showed normal male karyotype. He attained complete remission with aclarubicin and BH-AC and continued it until August 1987 when pancytopenia and hypoplastic bone marrow developed. Chromosome analysis of bone marrow cells showed normal male karyotype and gene analysis revealed germ-line configuration of breakpoint cluster region (bcr). Overt leukemia developed in May 1988 when his WBC count increased to 60, 600/microliters with 91% blasts, which were negative for peroxidase staining, positive for anti-Ia and CDw 41 by cell surface analysis, and positive for ultrastructurally demonstrable platelet peroxidase. A diagnosis of megakaryocytic leukemia was made. Chromosome analysis of bone marrow cells showed 46, XY, t(9;22) (q34;q11) and gene analysis revealed rearrangement of bcr. He died in November 1988. Our results and review of literature suggest that late appearing ph1 chromosome and rearrangement of bcr may occur in a variety of hematologic malignancies and influence the course of disease.

A 38-year-old male admitted to the Internal Medicine of Surugadai Nihon University Hospital, complaining of general fatigue and throat pain. The laboratory examinations revealed leukocytosis (83, 900/microliters) and an appearance of myeloblasts (90.2%) in the peripheral blood. The nucleated cell count was 56 x 10(4)/microliters with 85.5% myeloblasts in bone marrow. He was diagnosed as acute myeloblastic leukemia (AML). Though he received two courses of combination chemotherapy with daunorubicin, BH-AC, 6 MP and prednisone, one course of combination with mitoxantrone, etoposide and cytosine arabinoside and one course of combination with aclarubicin cytosine arabinoside and prednisone, he could not achieved remission. A chromosome analysis revealed 46, XY del(5)(q22). The amount of DNA fragments hybridized to 4.5 Kb v-fms probe in blastoid cells was approximately a half amount of normal persons. It is not defined the relationship between the decrease of fms and leukemia in this case. He was diagnosed de novo AML, since he had not been received the therapy with potential mutagenic and carcinogenic agents and had not been exposed the irradiation on his works.

A 56-year-old male was admitted to the Nihon University Hospital because of general fatigue and anemia on September 21st, 1985. He had mild hepato-splenomegaly. Hematological findings showed RBC 286 x 10(4)/microliters, Hb 6.0/dl, reticulocyte count 2.5%, platelet count 9.3 x 10(4)/microliters and WBC 2,400/microliters. An erythroblast per 100 leukocytes counted in a blood film was found. Bone marrow was erythroid hyperplasia with megaloblasts. The erythroblasts were PAS positive but not ringed sideroblasts. Other laboratory data including hemolysis were all negative. This case seemed to be diagnosed as refractory anemia (RA) according to the FAB classification. Chromosomal analysis of marrow cells, however, all revealed 46, XY, 20q- at diagnosis and 46, XY, 7q- 20q- after 22 months. Furthermore, Hb electrophoresis ahd family study indicated the presence of acquired HbH disease. Neither erythroid bursts (BFU-e) nor late erythroid progenitors (CFU-e) were detected. He has had progressive anemia without proliferation of blasts for over 2 years. From these findings, we postulate that the entity of erythremia should be distinguished from RA including many heterogeneous diseases.

Both the rearrangement and the expression of the bcl-2 gene in Japanese hematopoietic tumor cells were studied by Southern and Northern blot hybridizations. The expression of the bcl-2 gene was studied in seven cultured cell lines and twenty clinical samples, including eleven non-lymphoid tumors and nine lymphoid tumors. All lymphoid tumors except one sample from a patient with ALL expressed the bcl-2 gene. The bcl-2 gene was strongly expressed in B cell tumors. This gene was also expressed in T-ALL samples studied, although the expression was not as marked as in the B cell tumors. Non-lymphoid tumors did not express bcl-2 gene. bcl-2 gene rearrangement was studied in five cultured cell lines and six clinical samples including four follicular lymphomas and two T-ALLs. No abnormal bcl-2 gene configurations were found in any of the clinical samples. Among the cultured cell lines, the BALL-1 line showed two-fold amplification. The frequency of bcl-2 rearrangement in Japanese follicular lymphomas is reported to be lower than that seen in the American follicular lymphomas. Nevertheless, the increased expression of the bcl-2 gene seen in the Japanese B cell tumors studied supports the contention that the bcl-2 gene plays an important role in the pathogenesis of B cell tumors.

Carcinoembryonic antigen (CEA) gene was cloned in 1987. Thereafter, the structures of non-specific cross-reacting antigen (NCA) and biliary glycoprotein I (BGPI) have also been clarified. These three antigens contain immunoglobulin-like domains in their basic structures and it could be possible that CEA gene was originated from this basic structure by internal gene multiplication. Each CEA and NCA has hydrophobic domain in the C-terminus consisting of 26 amino acids which is eliminated when it binds with membrane and is reconstituted by combining with phosphatidylinositol glycan, whereas BGPI contains transmembrane and cytoplasmic domains. It is of interest that CEA and NCA have been found to function as adhesion molecules. The structure and possible function of another CEA gene family, PS beta G, were also introduced in this short review.

Epstein-Barr virus (EBV) infects human B lymphocytes and efficiently transforms them into immortalized lymphoblasts. EBV-determined nuclear antigen (EBNA) and EBV latent infection membrane protein (LMP) are expressed in latently infected, growth-transformed lymphoblasts. To elucidate the functions of EBNA and LMP, clones of cells were established that stably expressed EBNA-1, EBNA-2, EBNA-3A, EBNA-leader protein (EBNA-Lp) or LMP, using gene transfer technique and the growth characteristics of the transfectants were examined. The expression of EBNA-1, EBNA-2,EBNA-3A,EBNA-Lp or LMP caused shortening of doubling time, increased saturation cell density, reduced serum dependence, anchorage-independent growth in semisolid agar and activation of c-myc. Furthermore, the expression of LMP in NIH/3T3 cells led to tumorigenicity in nude mice, enhanced expression of H-ras and increased production of diacylglycerol, which might activate protein kinase C. B cell line, BJAB, EBNA-1 was responsible for expression of c-fgr mRNA and EBNA -2 specifically induced expression of B-cell activation antigens, including CD21 (CR2) and CD23 (Fc epsilon receptor). These results indicate that EBNA and LMP play an important role in EBV-induced growth transformation. It is possible that EBNA-1 and EBNA-2 are directly involved in the early process of immortalization. It is also possible that LMP could contribute to tumorigenic alteration of immortalized cells. The proliferation of the EBNA or LMP-expressing cells was markedly enhanced by phorbol ester. By contrast phorbol ester had no effect on the proliferation of nonexpressing control cells. The phorbol ester enhancement of EBV-induced growth transformation is likely to be mediated by EBNA and LMP.

The human papillomaviruses (HPVs) are epitheliotropic viruses that induce epithelial hyperproliferation including cutaneous warts and condylomas in cervical and vaginal epithelia. Certain types of HPVs, such as HPV 16, 18 are closely associated with cervical cancer, penile cancer and skin cancer of EV (epidermodysplasia verruciformis) patient. Experimental studies in cultured cell lines provide strong support for the idea that HPV 16 and 18 have transforming genes and are involved in the development of above cancers. However, their transforming genes are unable to induce malignant transformation of primary cultured cells originated from rodent, human for skin and cervical epithelial cells. For the carcinogenesis by HPVs, alteration of some additional factor(s) is thought to be necessary. It is the aim of this review to summarize recent knowledge about transforming genes of HPVs and their additional factors associated with carcinogenesis focused on cervical cancer.

Several genetic events are necessary for a cell to become malignant. Some of these events are activation of proto-oncogenes, and other events are loss of normal function of tumor suppressor gene(s). Two or more different tumor suppressor genes may be inactivated in some tumors, and the same suppressor gene may be involved in different types of tumors. Loss of constitutional heterozygosity (LOH) in the tumor suggests that a certain tumor suppressor gene may reside near the locus of the probe by which the LOH was demonstrated. However, LOH is not necessarily found when a tumor suppressor gene is inactivated. The frequency of LOH found by a probe depends upon the distance between the probe and the tumor suppressor gene. Moreover, inactivation of the suppressor gene by point mutation or very small deletion does not cause any change in electrophoretic mobility of the DNA fragment detected by the probe. Treatment of a cancer by tumor suppressor gene(s) is not possible until a technique by which we can introduce the gene into all the tumor cells is established. At present, clinicians should cooperate with basic scientists in the search for tumor suppressor genes. The comparison of clinical features and the genetic alterations in the tumor will shed light on the malignant behavior of the tumor cells such as rapid growth and/or tendency to metastasis.

Molecular biological analysis plays an important role for understanding of cancer pathogenesis. For example, accumulation of oncogene and anti-oncogene changes is necessary for cancer development. Molecular pathology is defined as a study field examining clinical materials using molecular biological technique. Even though mechanisms of cancer development and progression has much complexity, molecular pathological findings give a crucial information on clinical medicine, as Ki-ras point mutation and certain allelic loss may convert colon adenoma into carcinoma. Overexpression of EGF/receptor in gastric cancer is a biologic marker for high malignancy. More accumulation of molecular pathological informations will provide a new approach for prospective molecular diagnosis.

The patient was a 64-year-old woman who was admitted to our hospital because of lumbago. A diagnosis of multiple myeloma (non-producing type) was made, based on (1) the presence of multiple osteolytic lesions, (2) hypercellular marrow with 64.2% plasmacytoid malignant cells, (3) no monoclonal gamma-globulin was detected in the serum and urine, and (4) abnormal monoclonal gamma-globulin was also not detected in the cytoplasm and membranes of these malignant cells. After several courses of chemotherapy, a pleural effusion infiltrated by myeloma cells developed and the patient's serum contained a markedly increased amylase activity of salivary-type. Amylase activity was also detected in vitro in the supernatant of cultured myeloma cells established from the patient's pleural effusion. The presence of alpha-amylase in the myeloma cells, which were derived from pleural effusion, was demonstrated immuno-histochemically. These observations indicates that amylase was ectopically produced by these myeloma cells. Interestingly, 14 out of 20 metaphases in the cells derived from pleural effusion showed translocation of 1p22 near the region of 1p21, where the amylase gene was assigned.

We report a case of AML (M 4) with eosinophilia who developed meningeal relapse and transverse myelopathy. A 37-year-old woman was admitted to our hospital because of lymphadenopathy and ecchymosis. One week prior to admission, she noticed swelling of the cervical lymph nodes and bleeding tendency. On admission, low-grade fever, gingival swelling, generalized lymphadenopathy, and ecchymosis on the lower legs were found. A white blood cell count was 93,900/microliters with 82% blast cells, and a platelet count was 24,000/microliters. A bone marrow was composed of 45.3% myeloblasts, 27% monocytes and 7.1% eosinophils. Chromosome analysis revealed inv(16). The diagnosis of M4Eo was made. About one year after she gained complete remission, she was readmitted because of disturbance of urination. There was a sign of transverse myelopathy at the seventh vertebral level, and blast cells were detected in the cerebrospinal fluid. Despite of radiation and chemotherapy, paresis of lower extremities and sensory disturbance were persistent, and the patient died on 52th hospital day.

On November 22, 1985, a 54-year-old male was admitted to the cardiovascular department of our hospital suffering from thrombophlebitis, with redness, swelling and pain around the right ankle and left knee. He was transferred to our department on Nov. 26, because of hyperleukocytosis. Peripheral blood examination revealed hyperleukocytosis with 93.0% blastic cells and thrombocytopenia. The bone marrow aspirate was hypercellular and almost all cells were consistent with peroxidase negative blastic cells. The blastic cells were Leu M1 positive and Leu M2, M3 and lymphocytic markers were negative. The patient was diagnosed to have acute lymphocytic leukemia with Leu M1 positive blast cells. BH-AC/DMP therapy was began on the 1st hospital day but he died of cerebral haemorrhage on the 4th hospital day. An autopsy revealed systemic infiltration of leukemic cells including thrombophlebitis of the legs. Chromosome analysis of the bone marrow cells showed t(4;11)(q21;q23).

We studied the clinical and cytogenetic features of 14 acute lymphoblastic leukemia (ALL) patients with 1; 19 translocation. Ten patients had poor prognostic factors such as age over 10 years, hyperleukocytosis over 5 X 10(4)/microliters or high serum lactic dehydrogenase levels over 5,000 IU/l. Two patients had relapsed within 12 months after the onset, but their 5-year survival rate was 84.6%. Cytogenetically, 6 of 14 patients had multiple subclones. Two had the clones with hyperdiploidy greater than 50 chromosomes, which was known to be one of the favorable prognostic factors in childhood ALL. These findings show ALL children with 1; 19 translocation have a more favorable outcome in spite of some high-risk features than hitherto been thought.

We have evaluated the clinical efficacy and problematic items of flow cytometric surface and intracellular immunophenotyping of leukemic cells. The multi-parametric analysis on the results of leukemic phenotyping displayed a possibility of an independent phenotypical classification, but concurrently we recognized complementary relationships between the morphological and phenotypic approaches. Defective information on the morphology of leukemic cells unexpectedly introduced an insufficient quality in performance of leukemic phenotyping. On the other hand, the interpretations of phenotypic outcomes were complicated in cases with mixed leukemia and partial expression of a marker. Finally we hope further integration of the expression spectrum of markers, including the improvement of CD nomenclature system.

Xeroderma pigmentosum (XP) is an autosomal recessive human disease, clinically characterized by high incidence of skin cancers on sun-exposed areas. XP cells are hypersensitive to killing by ultraviolet light (UV), because they have a defect in DNA excision repair of UV-induced DNA damages. Genetic complementation analysis by cell fusion has identified 9 genetic complementation groups, designated groups A through H and a variant. However, the genetic basis of the physiological defect of XP has not yet been characterized. Recently, XP genes and human DNA repair genes have been molecularly cloned by DNA transfection methods. Molecular biological analysis of these genes should be a clue to elucidating the molecular mechanism of DNA repair in human. Moreover, an in vivo microinjection system and an in vitro system for study of DNA repair synthesis promoted by human cell extract have been developed and they can be utilized as assays during the purification of protein factors that complement repair defective XP cells. A nuclear factor that binds to DNA lesion has been identified and it was defective in group E XP cells. Yeast homolog of this nuclear factor appears to be a photolyase.

Adenoid cystic carcinoma generally consists of the following histologic features: tubular, cribriform, trabecular, and solid. To investigate how these histological patterns affect the prognosis of this carcinoma, we determined the proliferative activity of each of the histologic patterns by cytofluorometry. Twenty-six cases of adenoid cystic carcinoma, obtained by surgical resection, were studied. According to predominant histological pattern, they were divided into three groups: seven cases were of cribriform pattern, nine cases of trabecular pattern, and ten cases of solid pattern. The region with each dominant pattern was obtained from biopsy specimen, and the nuclear DNA contents of the tumor cells of the regions were assayed. In four of twenty-six cases, that of the tumor cells of the region with other patterns in the same specimens were also assayed. The results were the following: 1) The mean incidence of over 4.5C-polyploid cells of the region with predominant pattern of each tumor significantly increased in the following order: cribriform pattern and solid pattern. 2) The incidence of over 4.5C-polyploid cells, in comparison of a predominant pattern with other histologic patterns in the same tumor, were calculated. It was higher in solid pattern than in trabecular pattern, and was higher in trabecular pattern than cribriform pattern. 3) Only two of ten cases, had aneuploidy in the region with predominant solid pattern and the other cases had diploid stem line. Judging from the above results, it was concluded that proliferative activity of each pattern increased in the following order: cribriform pattern, trabecular pattern, solid pattern.(ABSTRACT TRUNCATED AT 250 WORDS)