A new human testicular cancer cell line (TCam-2) was established. The original material of TCam-2 was a primary lesion of a left testicular seminoma (typical pure type seminoma) from a 35 aged male patient. TCam-2 produced neither AFP nor beta-HCG, It showed strong immunoreactivities for 5G9 (anti testicular cancer MoAb), 4B3 (anti PTHrP MoAb) and PALP (placental alkaline phosphatase). The chromosomal analysis revealed 92 modal number and loss of Y chromosome. Histochemical, morphological and chromosomal analysis supported that TCam-2 is from classical seminoma. TCam-2 was transplanted subcutaneously to the back of 6 weeks old scid mice (CB-17 strain), and grew a classical seminomatous tissue.

Carcinogenesis is a multistep process in which cells accumulate multiple genetic alterations as they progress to a more malignant phenotype. Allelic chromosomal losses as well as mutations of tumor suppressor genes are common genetic events in the development and/or progression of both hereditary and non-hereditary tumors. Recent studies have shown a direct correlation between malignancy and inactivation of tumor suppressor genes for human cancers such as colorectal, hepatocellular and lung carcinomas. Although it is supposed that there are more than twenty tumor suppressor genes in the human genome, only a few tumor suppressor genes have been identified to date. Thus, further studies should focus on the identification and characterization of novel tumor suppressor genes, and molecular analyses of those genes in human cancer would be of great help in clarifying the multiple steps in the process of human carcinogenesis.

Telomere is the structure which is located on both ends of individual chromosomes in eukaryotes. The DNA sequence of the telomere consists of Guanine-rich tandem repeat, i.e., (TTAGGG)n in man. Telomere protects the end of the chromosome from fusion or deletion and maintains the stability of the chromosome and is synthesized by telomerase, a ribonucleoprotein. Telomere reduction is observed with cell senescence and immortalization, both in vivo and in vitro. Thus, telomere is considered to be a "clock" which measures the life span of cells, and its length is altered by cellular senescence and immortalization.

Three methods for analyzing the products of polymerase chain reaction were applied to detect complex alterations of dihydrofolate reductase (DHFR) gene, in order to assess their value in detection of folate-resistance in leukemia cells. A single point mutation in the second position of codon 31, a T-to-C transition, in trimetrexate (TMQ) resistant MOLT-3/TMQ200 cells was detected by either allele-specific oligonucleotide hybridization or restriction pattern of the PCR product. These two analyses allowed us to detect not only the presence of the mutation, but also the amplification of the mutated gene in TMQ-methotrexate (MTX) doubly-resistant MOLT-3/TMQ200-MTX500 cells. The base change was confirmed by direct sequencing method of the PCR product. Using these analyses of the PCR product, the complex alterations of DHFR gene are to be examined in leukemic patient cells.

Immunohistochemical staining for EGF, EGFR, c-erbB-2, p53, K-ras and PCNA was performed on the formalin-fixed, paraffin embedded sections of resected gastric carcinomas. A relatively high positive rate was observed for EGFR and c-erbB-2 in the well-differentiated adenocarcinomas and p53 in the poorly-differentiated adenocarcinomas. The positive rate of these factor was higher in the advanced cases than in the early cases, and also in the deep invasive area than the superficial area. According to the PCNA staining, a relatively high positive rate was observed in the well-differentiated adenocarcinomas compared with the early cases of poorly-differentiated adenocarcinomas, but the positive rate was markedly higher in the advanced cases of the latter. Typical signet-ring cell carcinomas showed the lowest positivity rate compared with the other histological types of gastric carcinomas.

Cultured human retinoblastoma cells (Y79) were induced to differentiate by dibutyryl cyclic AMP(Bt2cAMP). We examined the effects on the mRNA levels of several cellular genes when the induction of differentiation was monitored by observation of the cellular processes. Bt2cAMP(1mM) treatment produced significant extension of cellular process after 3 days. We examined the mRNA levels of N-myc gene(oncogene), Rb(anti-oncogene, retinoblastoma gene) and nucleolin (nucleolar protein) being linked with ribosome biosynthesis. The mRNA levels of all these genes decreased for 3 days after Bt2cAMP treatment. These results suggested the possibility that Y 79 cells were induced to differentiate by down-modulation of both N-myc gene expression and ribosome biosynthesis in the nucleolus following treatment of Bt2cAMP. Furthermore, the gene expression of the retinoblastoma gene is likely to be downregulated by this condition even if the product of mRNA is not functional.

We investigated the significance of HPV 16, 18 infection and DNA ploidy in the progression to malignancy of uterine cervical dysplasia. Formalin-fixed, paraffin-embedded surgical and biopsy specimens obtained from 59 women were examined. The 59 women were classified into two groups: one of progression cases and the other regression cases. Polymerase chain reaction (PCR) and dot blot hybridization (DBH) were used for the detection of HPV 16/18 DNA, while flow cytometry was used for the analysis of the relative cellular DNA content in samples prepared by Hedley's method. Eight out of 29 progression cases (27.6%) and 3 out of 30 regression cases (10.0%) exhibited HPV 16/18 DNA, whereas 14 out of 29 progression cases (48.3%) and 2 out of 30 regression cases (6.7%) involved a DNA aneuploid population, representing a DNA index in the former above 1.5 and in the latter below 1.5. The proportion of cases having a population of DNA aneuploid together with the presence of HPV 16/18 DNA was 20.7% (6/29) in the progression group and 0% (0/30) in the regression group. These results indicate that the DNA aneuploid population coupled with simultaneous HPV 16, 18 infection may be used as a marker for progression in uterine cervical dysplasia.

[Case 1] A 44-year-old female was referred to our hospital because of leukocytosis. The WBC count was 26400/microliters and NAP score 21. As Ph1 chromosome was detected, she was diagnosed as CML and treated with busulfan. Because of the rapid decrease of WBC, we stopped busulfan. Progressive pancytopenia and an increase of myeloblasts and promyeloblasts in the bone marrow was observed. We started vincristine and prednisolone therapy. Ph1 chromosome was not detectable and southern blot analysis did not show rearranged bands of M-bcr three years after the last therapy. [Case 2] A 74-year-old female was referred to our hospital by reason of leukocytosis and thrombocytosis. The WBC count was 22,500/microliters, the platelet 907,000/microliters, NAP score 53, and Ph1 chromosome was found. The diagnosis of CML was made, and she was treated with busulfan. The WBC rapidly fell to 1,900/microliters, when chromosome analysis revealed the presence of Ph1 negative clones (4/20). She was admitted due to thrombocytopenia and leukocytosis with the additional chromosome change of i (17q). Her peripheral blood and bone marrow pictures were consistent with blast crisis, and she died of cardiac tamponade. These two cases show the heterogeneity of CML patients, and also suggest the possibility that keeping the WBC count low may lead to a decrease of Ph1 positive clones.

It has been shown that molecular cytogenetics in interphase nuclei is applicable to paraffin embedded sections from formarin-fixed materials utilizing in situ hybridization (ISH). This allows precise identification of numerical abnormalities of chromosome in tumor cells without disruption of tumor morphology. Thirteen cases of bladder cancer and twenty six cases of prostate adenocarcinoma were evaluated for numerical aberrations of chromosome 1, 7, 10, 11, 17, 18, X and Y utilizing biotinylated probes specific for the alpha satellite region. In two cases from total prostatectomy, one case from total cystectomy and two cases from TUR-P specimen, hybridized chromosomes could not be detected in individual tumor cells. With respect to the appearance of the ISH signal, optical concentration and time for the digestion enzyme has to be established essentially. This technique can now be applied on the detection of biological activity in various types of urological cancer.

Correlation between histological type, depth of invasion, nodal involvement, macroscopic type, survival rate and nuclear DNA content was studied by flow cytometer using paraffin-embedded materials from 52 cases of superficial esophageal carcinoma. There were 23 cases (44.2%) of diploid and 29 cases (55.8%) of aneuploid. There was no correlation between DNA aneuploidy and depth of invasion or size of the carcinoma. Carcinoma with aneuploid had a high frequency of nodal involvement and well-differentiated histological type. Carcinoma with deep of highly elevated macroscopic type has a tendency to have higher DNA content. There was no correlation between survival rate and nuclear DNA ploidy patterns.

A human ovarian cancer cell line designated "MH" was established from ascites of a patient with ovarian serous cystadenocarcinoma treated with cisplatin. This cell line was grown for more than 2 years and over 100 passages in medium RPMI1640 containing 10% FCS. The cell doubling time, saturation density and plating efficiency was approximately 6.3 days, 5.5 x 10(4)/cm2, and 42%, respectively. Chromosomal analysis revealed aneuploidy with a modal number of 72 and 14-19 types of marker chromosomes. CA125 was detected in both the original tumor and the cultured cells. This cell line is cisplatin-resistant (IC50: 3.28 microM) and has high protein kinase C activity.

An 83-year-old male was admitted with a right pleural effusion and generalized lymphadenopathy. Serum LDH level was elevated to 801 IU/L, and the pathological diagnosis from inguinal lymph node needle biopsy was malignant lymphoma (ML) of diffuse, large cell, non-cleaved type, according to the working formulation. The surface phenotypes of the malignant cells from the pleural effusion were analyzed by a fluorescent-activated cell sorter with a panel of monoclonal antibodies (MAbs). The ML cells coexpressed antigens detected by MAbs CD10 (CALLA), CD19, CD20, CD22, CD24, CD38, Ia, c-neu and surface immunoglobulin G kappa. A high expression of NRAS p21 was also detected by cytoplasmic immunofluorescence technique. The patient died 19 days later despite a combination of chemotherapy and intensive supportive therapy. From these findings it seems that c-neu may be a prognostic indicator not only for breast cancers but also for lymphoproliferative disorders. Further accumulation of such cases is needed.