Hidenobu Ogata

Comparative genomic hybridization detects genetic alterations during early stages of cervical cancer progression.

Invasive cervical carcinoma is thought to arise from cervical intraepithelial neoplasm (CIN). Genetic changes that occur during progression of CIN to cervical carcinoma are poorly understood, although they appear to be directly involved in this process. We used comparative genomic hybridization (CGH) with precise microdissection and degenerate oligonucleotide primed‐polymerase chain reaction (DOP‐PCR) to detect genetic alterations in normal epithelial, CIN, and invasive carcinoma tissues colocalized in tumors from 18 patients with squamous cell carcinoma of the uterine cervix. Gains on chromosome 1 and on 3q and losses on 2q, 3p, 4, 6p, 11q, and 17p were frequent alterations found in CIN and invasive carcinoma lesions. Interestingly, several of these genetic changes were observed in preinvasive carcinoma lesions. The frequency and average number of genetic alterations corresponded directly to the extent to which the cervical carcinoma had progressed. Frequent alterations were found in more than 90% of CIN III lesions. Gains on 3q and losses on 11q were the most prevalent genetic alterations found in association with uterine cervix carcinogenesis. The common regions of alteration were 3q26.1‐q28 and 11q23‐qter. The majority of tumor samples showed variability in genetic alterations across lesion types within a single specimen.

Genetic aberrations detected by comparative genomic hybridization predict outcome in patients with endometrioid carcinoma.

Endometrial cancer progression is determined by a complex pattern of multiple genetic aberrations, but how these aberrations affect prognosis is unknown. In this study, we undertook a genome‐wide screening to detect genetic changes by comparative genomic hybridization (CGH) in 51 tumors from patients with primary endometrioid carcinoma of the uterine corpus. The observed genetic changes were subsequently correlated with the progression of the disease and the clinical outcome in each case. The average number of genetic aberrations (copy number gains and losses) was significantly greater in non‐surviving patients than in disease‐free patients (12.6 vs. 2.7, P < 0.0001). According to multivariate analysis, lymph node metastasis (P = 0.015), cervical involvement (P = 0.007) and one or more copy number losses at 9q32–q34, 11q23, or Xq12–q24 (P = 0.023) were significantly predictive of death from the disease. Interestingly, lymph node metastasis was significantly associated with copy number gains at 8q22–q23 and 8q24–qter (P = 0.003 and P = 0.025, respectively). Moreover, cervical involvement was also correlated significantly not only with gains of 8q22–q23 and 8q24–qter but also with loss of 11q23 (P = 0.04, 0.0003, and P = 0.009, respectively). These results suggest that analysis of genetic changes may help predict clinical outcome and the presence of metastatic disease as well as assist in therapeutic decision making for patients with endometrioid carcinoma. Genes Chromosomes Cancer 29:75–82, 2000.

Pelvic exenteration for the treatment of gynecological malignancies

Twenty-three patients undergoing pelvic exenteration for primary and recurrent gynecological malignancies from 1976 to 1994 are reported. Fifteen patients underwent total pelvic exenteration, 3 underwent anterior exenteration, and 5 underwent a posterior procedure. Eight patients had exenteration as their primary treatment (primary group), and 15 underwent exenteration as secondary treatment (recurrent group). In the primary group, two patients developed recurrence and died of it at 6 and 20 months after operation. Five patients are still being followed up and are alive without disease. Four of these 5 patients have survived more than 5 years. In the recurrent group, 12 patients were followed up and three died of complications during the early years. Seven patients died of cancer with the mean survival time of 16.6 months. The mean age, average operating time, and mean blood loss in the primary and recurrent groups were 57 vs. 53 years, 8 hours and 20 min vs. 8 hours and 10 min, and 4,120 vs. 4,190 ml, respectively. The overall cumulative 5-year survival rate was 34.7%, being 68.6% in the primary group and 16.7% in the recurrent group. It is noteworthy that the 5-year survival rate was 51.3% in the patients who had surgical margins free of disease. In conclusion, pelvic exenteration should be considered an acceptable therapeutic option when appropriately selected.

Identification of squamous cell carcinoma antigen-2 in tumor tissue by two-dimensional electrophoresis.

The aim of this study was to identify two homologous serine proteinase inhibitor (serpin) molecules, squamous cell carcinoma (SCC) antigen‐1 and ‐2, by two‐dimensional electrophoresis (2‐DE), combined with immunoblotting, and examine their expression in tumor tissue. The recombinant SCC (rSCC) antigen‐1 showed four spots with pI 6.5, 6.4, 6.3 and 6.0, whereas rSCC antigen‐2 showed a more acidic spot with pI 5.95. SCC antigen in tumor tissue appeared in three new acidic spots (pI 5.7—5.5, Mr 44 500), numbered 5, 6 and 7, besides the previously reported four spots numbered 1 to 4. These new acidic spots of SCC antigen apparently increased in SCC tissue. Treatment of tissue extract by carboxymethyl (CM)‐papain agarose matrix extinguished spots 1 to 4 encoded on the SCCA1 gene, but not 5 to 7 on the SCCA2 gene. Overexpression of the SCCA2 gene may play an important role in the malignant behavior of tumor cells.

Nondenaturing two‐dimensional electrophoretic analysis of loop‐sheet polymerization of serpin, squamous cell carcinoma antigen‐2

Two homologous serine proteinase inhibitors (serpins), squamous cell carcinoma (SCC) antigen‐1 and ‐2 were separated by nondenaturing two‐dimensional electrophoresis combined with immunostaining to acquire further information on these proteins under physiological conditions. Polymers of SCC antigen‐2 were detected in cytosolic extracts prepared from tumor tissues. The polymer formation of SCC antigen‐2 was apparently decreased and the SCC antigen‐2‐synthetic peptide binary complexes were newly formed by the addition of synthetic peptide with sequences corresponding to residues from P14 to P2 in the reactive center loop of SCC antigen‐2. On the other hand, the incubation with synthetic peptides having the sequence of the reactive center loop of SCC antigen‐1 or antithrombin had no effect on polymerization of SCC antigen‐2. These data suggest that the polymerization of SCC antigen‐2 may occur spontaneously in vivo by the loop‐sheet mechanism of serpin.