Interpreting microarray data: towards the complete bioinformatics toolkit for cancer.

Functional genomics has been applied in the study of human malignancies since the inception of this field nearly a decade ago. Microarray analysis has been specifically used in an attempt to re-classify carcinomas at the molecular level, to aid in diagnosis/prognosis and to predict how various types of tumour respond to different therapeutic agents. Bioinformatics is now at the forefront of the post-genomics era and is providing a number of tools with which to mine the large datasets produced by genome-wide analysis.

Human papillomaviruses in cervical cancer: II. Application of PCR and in situ hybridization and HPV significance.

Human papillomaviruses especially type 16 and 18 are associated with cervical cancer. These viruses encode transforming proteins which are capable of binding and form complexes with p53 protein. Tumour-tissue samples from women with cervical cancer as well as normal controls were analysed for the presence of HPV and the expression of p53 by in situ hybridization and PCR analysis.

Human papillomaviruses in cervical cancer I. HPV-16 and 18 predominate in the Greek population.

Human papillomaviruses (HPV) and their role in carcinogenesis have been the subject of extensive investigation Specific types of HPV have been associated with cervical carcinoma HPV 16 and 18 are mainly associated with malignant progression and considered "high risk" viruses Using Southern blot analysis and in situ hybridization we investigated the presence of papilloma viruses in cervical carcinoma patients as well as appropriate controls. The results presented here support the aetiological role of HPV 16 and 18 in cervical carcinoma and demonstrate the prevalence of these viruses in Greek women. The role of viruses in carcinogenesis in well established in almost all species from fishes, to birds, to mammals.

Expression of ets family of genes in systemic lupus erythematosus and Sjogren's syndrome.

We have examined the expression of three members of the ets family of genes, ETS1, ETS2 and ERGB/Fli-1, in lymphocytes from patients with systemic lupus erythematosus and in two murine autoimmune model systems. The ERGB/Fli-1 gene is expressed at a higher level in lymphocytes from autoimmune disease patients than healthy individuals. In addition, we found that the ERGB/Fli-1 gene expression is higher in splenic T-cells from lupus prone mice and in infiltrating lymphocytes in the salivary glands of HTLV-I tax transgenic mice.

Heat turn-over during hyperthermic peritoneal perfusion. An experimental study.

In order to study heat turnover, 3 experimental models of chemotherapeutic hyperthermic intraperitoneal perfusion (CHIP) were tested, using 15 rabbits divided into 3 separate groups of 5 animals each. A normal saline perfusate, containing a standard concentration of 10 mg of methotrexate per liter, was recycled through the peritoneal cavity, transmitting, in every group, 6,000 calories of heat. In model I, heat transmission was achieved by a high temperature gradient, in model II by the increased thermocapacity of a great priming volume of perfusate, using a peritoneal expander, and in model III by a high flow rate.

Nasopharyngeal carcinoma: Epstein-Barr Virus significance.

Thirty-one cases of NPC were investigated by in situ hybridization, PCR analysis and serology to assess the presence of EBV and to interrelate these findings. Statistical analysis showed a significant correlation between the probability of detecting EBV and NPC. There was no significant association between in situ hybridization, PCR and serology.

Establishment and characterization of a hepatoblastoma cell-line from a patient with secondary acute myeloid-leukemia.

A girl who had been treated, apparently successfully, with surgery and chemotherapy for a hepatoblastoma, fell ill two years later with what was diagnosed as an AMF M(4). A cell line was established from her peripheral blood. This cell line had epithelial morphology and grew both in suspension culture and as a monolayer.

P53 gene mutations in women with breast cancer and a previous history of benign breast disease.

Mutations of the p53 tumour suppressor gene are the most common genetic lesions in human cancers and have been reported in breast cancer as part of the Li-Fraumeni syndrome. In the present study, we determined frequencies and types of the p53 mutations in breast cancer tissues in women with a history of benign breast disease (BBD) identified in Florence, Italy, with (n = 6) or without (n = 10) a family history of breast cancer. Among the cases with a family history of breast cancer and BBD, 2 out of 6 had p53 gene mutations in cancer samples.

Regional intraarterial infusion of methotrexate with leucovorin rescue in a rat tumor model.

Walker 2S6 carcinosarcoma was transplanted in the paw of 52 male Wistar rats. A week later, the animals were divided into 6 separate groups and treated in different ways, as follows. Group I (control): no treatment.

Relative paucity of p53 gene-mutations in male breast carcinomas.

Mutations of the p53 suppressor -ene are the most common genetic lesion noted in human cancers and appear to be relatively common (30%) as somatic cell mutations in female breast cancer. p53 mutations have also been frequently reported in familial breast cancers as in Li-Fraumeni syndrome (LFS). Males with breast cancer are far rarer than females.

Subtraction hybridization cDNA libraries from colon carcinoma and hepatic cancer.

cDNA clones of differentially expressed mRNAs in a colon carcinoma and a hepatocellular carcinoma have been isolated by subtractive cDNA cloning. The subtracted material is at least 90 X enriched for differentially expressed sequences and can be used for construction of subtractive cDNA libraries and polymerase chain reaction (PCR) amplification to generate differential probes. Commercially available lambda ZAP II is used for construction of primary libraries since single-stranded phage bearing the cloned cDNA can be excised in vivo and because lambda libraries are convenient for subsequent screening and manipulations.

Herpes viruses and duodenal ulcer disease.

Antibodies to herpes simplex virus and Epstein-Barr virus were sought in the sera of 33 patients with endoscopically proved duodenal ulcer and 33 healthy controls. Neither the prevalence nor the antibody levels to both viruses were significantly different between the two groups. These results suggest that herpes viruses may not play a role in the pathogenesis of duodenal ulcer disease.

Combination of hyperthermia and methotrexate in the treatment of transplanted Walker sarcoma.

The purpose of this experimental study was to evaluate the possible synergistic effect of hyperthermia (HPT) and methotrexate (MTX) in Wistar rats transplanted with Walker sarcoma. HPT was induced by microwaves of 432 MHZ frequency transmitted through a source of 2 x 5.5 cm2 surface.

Establishment and characterization of a B-cell line from a patient with acute lymphoblastic leukemia.

A permanent lymphoblastoid cell line was established from the peripheral blood of a child with acute lymphoblastic leukemia. The cell line, designated SDK, grows in a stationary suspension culture, forming aggregates, in RPMI medium supplemented with 10% FCS, with a doubling time of 50-60 h. Immunologic markers and cytological features suggested that the SDK cells should be identified as being of B-cell origin.

Interactions between human fibroblasts and HeLa cells in vitro.

Affinity toward each other was demonstrated in co-cultures between HeLa cells and fibroblasts originating from human tumor stromal or normal tissues. Both cell types in the mixed cultures (ratio 1:1, 1:2, 2:1) proliferated normally as shown by 3H-thymidine labeling index estimation for up to 48 hr of co-culture. At ratios of fibroblasts: HeLa lower than 1:10, fibroblasts were eventually eliminated after serial passaging.

Uncontrolled growth of tumour stromal fibroblasts in vitro.

The disturbance of the growth control mechanisms in tumour cells in vivo may be manifested as uncontrolled growth of the tumour stroma in vitro. Stromal fibroblast-like finite cell lines produced from benign or malignant human breast tissue specimens exhibited cell overlapping which ranged from multilayers to dense piling up colonies, while cells derived from normal tissues exhibited intense contact inhibition of growth and locomotion, under the same culture conditions. 6 out of 13 lines derived from malignant tissues and 2 out of 5 lines derived from benign lesions exhibited the phenomenon of 'periodic appearance of piling up colonies'.

Interactions between human lymphoblastoid cells and human fibroblast feeder layers in vitro.

Lymphoblastoid (LB) cells interact in vitro with human fibroblasts. Cell complexes can be dissociated by trypsin treatment. Binding on fibroblasts is higher for Epstein-Barr virus-producer LB cells than for nonproducers, and depends on the origin of the fibroblastic cells.