Daisuke Doi

Novel gene fusion of COX6C at 8q22–23 to HMGIC at 12q15 in a uterine leiomyoma

Cytogenetic analyses have shown that aberrations involving 12q13–15 are frequent chromosomal changes in a variety of human benign mesenchymal tumors, e.g., pleomorphic adenomas of the parotid gland, pulmonary chondroid hamartomas, lipomas, and uterine leiomyomas. Recently, the high‐mobility group protein gene HMGIC was identified as the target gene affected by the 12q13–15 aberrations. Using 3′ rapid amplification of cDNA ends experiments, we isolated novel ectopic sequences fused to HMGIC in a uterine leiomyoma. Cloning of the fusion cDNA identified the human cytochrome c oxidase subunit VIc (COX6C) gene on 8q22–23 as the fusion partner of HMGIC. Nucleotide sequences of the fusion transcript revealed that the first 3 exons of the HMGIC gene, encoding the 3 DNA binding domains, was fused to the exon 2 of the COX6C gene. The identification of a gene rearrangement suggests a role for HMGIC in tumorigenesis of uterine leiomyoma and suggests a possible involvement of HMGIC in mesenchymal differentiation. Genes Chromosomes Cancer 27:303–307, 2000.

Gene fusion involving HMGIC is a frequent aberration in uterine leiomyomas

AbstractHMGIC, a high-mobility-group protein gene encoding an architectural transcription factor, was recently identified as the target of gene fusion in a variety of human benign mesenchymal tumors; some of these events were chromosomal translocations involving 12q13–15. HMGIC consists of three DNA-binding domains (encoded by exons 1–3), a spacer, and an acidic carboxyl-terminal regulatory domain (exons 4–5). To determine the spectrum and nature of the aberrations in uterine myomas in Japanese patients, we systematically examined the tumors of 45 patients for all possible types of gene fusions involving HMGIC, by means of 3′-rapid amplification of cDNA ends (RACE) and reverse transcriptase-polymerase chain reaction (RT-PCR) experiments. HMGIC gene fusions were found in 16 (36%) of the tumors; aberrant splicings to five cryptic sequences located in introns of the HMGIC gene were found in 11 of these cases, and translocations causing juxtaposition to other genes, such as COX6C and RAD51B, were found in 5. In all fusion transcripts, the first two or three exons of HMGIC were fused to ectopic sequences. Our results suggest that a fusion event, resulting in the separation of the DNA-binding domains of HMGIC from the spacer and the acidic carboxyl-terminal regulatory domain, is a common tumorigenic mechanism in the development of uterine myomas.

Serum adenosine deaminase activity and its isoenzyme pattern in women with normal pregnancies

Abstract. Adenosine deaminase (ADA) is a purine enzyme which is essential for the proliferation, maturation and function of lymphoid cells, and congenital deficiency of this enzyme is associated with severe combined immunodeficiency disease. The activity of ADA has changed in diseases characterized by the alteration of cell-mediated immunity such as rheumatoid arthritis, systemic lupus erythematosus and tuberculosis, so ADA has been considered as a nonspecific marker of cell-mediated immunity. In this study we examined changes in serum total ADA activity and the patterns of two ADA isoenzymes, ADA1 and ADA2 in normal pregnant women, and evaluated the possible role of the alteration of cell-mediated immunity during normal pregnancy as causes of changes in ADA activity. We measured serum activities of total ADA, ADA1 and ADA2 in normal pregnant women in the third trimester (n=24) and age-matched healthy nonpregnant women (n=24). Peripheral blood lymphocytes and monocytes were also measured. In normal pregnant women, serum total ADA activity averaged 10.5 ± 0.5 U/L, which was significantly lower than in nonpregnant women (14.0 ± 0.5 U/L ) (p<0.05), and mean serum ADA2 activity also significantly reduced that of nonpregnant women (p<0.05). There was no significant difference in ADA1 activity in normal pregnant and nonpregnant women. The decrease in total ADA activity was accompanied by the decrease in lymphocyte count. These results suggest that reduced serum total ADA activity reflects decrease in ADA2 activity, and which may be in part associated with depressed cell-mediated immunity during normal pregnancy.

Serum Adenosine Deaminase Activity in Women with Pre-Eclampsia

The present study investigated serum adenosine deaminase (ADA) activity and the patterns of two ADA isoenzymes, ADA1 and ADA2, and to evaluate the possible role of cell-mediated immunity as causes of the changes in ADA activity in pre-eclampsia. We measured serum activities of total ADA, ADA1 and ADA2 in pre-eclampsia (n = 22) and normal pregnancy (n = 22). Peripheral blood monocyte counts and neopterin levels, reflecting the activation of the monocyte-macrophage cell system, were also measured. In pre-eclampsia, serum total ADA and ADA2 activities were significantly increased compared with normal pregnancy (p < 0.05), which were accompanied by increases in serum neopterin levels. These results suggest that increased total ADA activity reflects increases in ADA2 activity, which may be in part related to enhanced cell-mediated immunity during pre-eclampsia.

Significance of cervical gland area in predicting preterm birth for patients with threatened preterm delivery: comparison with cervical length and fetal fibronectin.

Background/Aims: Absent cervical gland area (CGA) has been considered a predictor of preterm delivery (PTD) for women at low risk. Predictive efficacy was analyzed in women at high risk for PTD and compared with cervical length (CL) <20 mm and fetal fibronectin (fFN) in cervicovaginal secretions. Methods: Case notes were reviewed for 108 subjects with gestation of 22–33 weeks who had been admitted to hospital with threatened PTD. The uterine cervix was observed by vaginal sonography and fFN was sampled on admission. Relationships between findings and outcome of PTD at <34 weeks’ gestation were analyzed. Results: Delivery at <34 weeks’ gestation occurred in 14.8% of patients. Absent CGA (68.8%), short CL (75.0%), short CL without CGA (62.5%) and positive fFN (62.5%) were more frequent in these patients than in patients undelivered at <34 weeks’ gestation (p < 0.05). Logistic regression analysis identified positive fFN and short CL with absent CGA as independent predictors for PTD (p < 0.0001). The mean interval from admission to delivery was 2.9 weeks in cases with fFN and both sonographic findings, compared to 9.3 weeks in cases with fFN but both sonographic finding (p = 0.0005). Conclusion: Short CL with absent CGA represents an independent predictor for PTD, as does fFN.

Relationship between adenosine deaminase activity and cytokine-secreting T cells in normal pregnancy.

OBJECTIVE To evaluate the relationship between plasma adenosine deaminase activity and the proportion of cytokine‐secreting T cells as causes of changes in adenosine deaminase activity in normal pregnancy. METHODS Plasma adenosine deaminase activity and the proportions of cytokine‐secreting T cells were measured in the peripheral blood of 26 nonpregnant and normal pregnant women in the third trimester. The proportion of CD4‐positive T cells secreting interferon‐γ derived from T helper 1 cells, and interleukin‐4 derived from T helper 2 cells, were analyzed by flow cytometry. The ratio of interferon‐γ–secreting cells to interleukin‐4–secreting cells was taken as the T helper 1/T helper 2 ratio. RESULTS Mean plasma adenosine deaminase activity in normal pregnant women, averaged, was significantly lower than that in nonpregnant women (10.3 ± 0.6 U/L versus 13.8 ± 0.5 U/L, P < .05). In normal pregnant women, the proportion of interferon‐γ–secreting cells was significantly lower than that in nonpregnant women (20.5% ± 1.0% versus 24.7% ± 1.2%, P < .05), but the proportion of interleukin‐4–secreting cells did not differ from that of nonpregnant women. Consequently, the T helper 1/T helper 2 ratios were significantly decreased during normal pregnancy. A significant correlation was found between adenosine deaminase activity and the proportion of interferon‐γ–secreting cells (r = .54, P < .05). CONCLUSION Decreased plasma adenosine deaminase activity in normal pregnant women may be in part associated with changes in the immunological status, especially the decrease of the proportion of interferon‐γ–secreting cells.

Changes in the proportion of T helper 1 and T helper 2 cells in cord blood after premature rupture of membranes

Abstract. This study investigated changes in the proportion of T helper (Th)1 and Th2 cells in cord blood after premature rupture of membranes (PROM), and evaluate the effects of PROM on the intrauterine fetal immune status. The proportion of CD3-positive T cells secreting interferon (IFN)-γ as an index of Th1 cells, and interleukin (IL)-4 as an index of Th2 cells in cord blood of 12 newborns with and without PROM, were analyzed by flow cytometry. In cord blood of newborns with PROM, the proportion of IFN-γ secreting cells significantly increased, and the proportion of IL-4 secreting cells was rather high but not significantly higher than that of newborns without PROM. These changes eventually caused a shift in the Th1/Th2 ratio to Th1 dominance in PROM. There was no significant correlation between the proportion of IFN-γ secreting cells and the duration of PROM before the onset of labor. These results suggest that the increase in the proportion of IFN-γ secreting cells after PROM, which eventually cause the Th1/Th2 ratios to show the Th1 predominance, may reflect in part intrauterine fetal immune responses to PROM.

Plasma 5′-Nucleotidase Activities and Uric Acid Levels in Women with Pre-Eclampsia

The present study investigated plasma activity of 5′-nucleotidase, a key enzyme in the production of adenosine, in pre-eclampsia, and evaluated the relationship between changes in 5′-nucleotidase activity, and levels of uric acid, endproduct of the purine metabolism, and the severity of pre-eclampsia. We measured plasma 5′-nucleotidase activities and uric acid levels in women with 18 normal pregnancies, mild and severe pre-eclampsia. In mild and severe pre-eclampsia, plasma 5′-nucleotidase activities and uric acid levels were significantly increased compared with those in normal pregnancy (p < 0.05). Plasma 5′-nucleotidase activity increased according to increases in uric acid levels and the severity of pre-eclampsia. These results suggest that increased plasma 5′-nucleotidase activity may, at least in part, be related to changes in purine metabolism in pre-eclampsia.

Immunohistochemical Localization of Tenascin, Estrogen Receptor and Transforming Growth Factory-β1 in Human Endometrial Carcinoma

Tenascin is an extracellular matrix glycoprotein which plays a role in cell attachment, proliferation and migration. To elucidate the function of tenascin in the proliferation of endometrial carcinoma, we studied tenascin expression in the endometrial carcinoma of 36 cases. In 22 of the carcinomas, tenascin expression was intense in the entire extracellular space, especially at the front of muscle invasion. Furthermore, in cases with metastases, deep invasion into muscles and vascular invasion, the rate of tenascin expression was significantly high. Immunoelectron microscopy revealed the tenascin reaction product in the stroma around fibroblasts located some distance from the basal lamina of cancer cells. On the other hand, tenascin expression was found in a high proportion of cases showing weak or no expression of estrogen receptor, and intense expression of transforming growth factor-beta 1. These results suggest that tenascin not only promotes cell proliferation and invasion but also inhibits further proliferation of carcinoma.

Serum CA 125 Level after Neoadjuvant Chemotherapy is Predictive of Prognosis and Debulking Surgery Outcomes in Advanced Epithelial Ovarian Cancer

Recently, neoadjuvant chemotherapy followed by interval debulking surgery (NACT-IDS) has been recommended for selected patients with International Federation of Gynecology and Obstetrics (FIGO) stage III or IV disease and bulky tumors. The aim of this study was to evaluate associations between post-NACT serum CA 125 levels, surgical outcomes, and clinical outcomes in patients with advanced epithelial ovarian cancer. We retrospectively analyzed 107 patients with FIGO stage III or IV ovarian cancer who were treated with NACT-IDS at the Gynecology Department of Kanagawa Cancer Center between January 2001 and December 2012. Serum CA 125 levels after NACT were significantly lower in the complete/optimal IDS group compared to the suboptimal IDS group (mean±standard deviation: 48.1±27.6 vs. 346.5±295.2 U/mL, p<0.01). Patients with low preoperative CA 125 levels (<35 U/mL) had a higher probability of optimal IDS (78.1±41.9% vs. 33.3±19.2%, p<0.01) and longer progression-free survival (mean±standard deviation: 30.4±14.3 months vs. 21.3±7.3 months, p<0.05) than patients with high CA 125 levels (>100 U/mL). Patients with low CA 125 levels (<35 U/mL) had a higher probability of complete/optimal IDS and longer progression-free survival compared to patients with high CA 125 levels (>100 U/mL).