Nihon Sanka Fujinka Gakkai zasshi最新文献

Introduction: Studies of tumor invasion and metastases have focused on the degradation of the basement membrane, which is predominantly made up of type IV collagen, laminin, and heparan sulfate proteoglycans. Matrix metalloproteinase-2 (MMP-2) and MMP-9, which can degrade type IV collagen, are implicated in cancer invasion and metastasis. Released and activated MMPs are controlled by specific tissue inhibitors of metalloproteinase (TIMP). In the present study, we have examined gelatinolytic and TIMP activity in the conditioned medium of human normal and cancer tissues by zymography and reverse zymography.

Materials and methods: 1) Tissues. Tissues were obtained at operation after informed consent was got from each patient. Sliced tissues were incubated in serum-free medium for 4 or 24 h at 37 degrees C. Human ovarian cancer cells (SAOV) were cultured for 24 h in serum-free medium containing conditioned medium of stromal tissues. After washing by PBS 3 times, SAOV cells were cultured for a further 24 h. 2) Zymography. Conditioned medium was subjected to SDS polyacrylamide gel containing 0.3 mg/ml of gelatin in zymography, and purified MMPs were added further in reverse zymography. After electrophoresis the gel was washed with Triton X-100, and incubated for 20 h at 37 degrees C in the reaction buffer. The gel was then stained with Coomassie brilliant blue. The gelatinase and TIMP activities were detected as unstained and stained bands, respectively. The photographs of the gels were scanned with a densitometer. 3) Other method. TIMP-1 levels of conditioned medium were assayed by ELISA kit. 4) Statistics. Statistical comparisons were made by Mann-Whiteny U test.

Results and discussion: We have examined the gelatinolytic activity in gynecologic normal and cancer tissues by zymography and reverse zymography. Ovarian, cervical, and endometrial cancer tissues demonstrated higher gelatinolytic activity than normal tissues. The major gelatinases were those with molecular weight of 92 and 72kD, which corresponded to MMP-9 and MMP-2, respectively. The ratio of MMP 9 to MMP-2 was significantly higher in 3 types of cancer tissues than in normal tissues. Reverse zymography demonstrated that TIMP-1 and TIMP-2 were present in all tissues, and the ratio of TIMP-1 to TIMP-2 was significantly higher in 3 types of cancer tissues than in normal tissues. These findings suggested that MMP-9 and TIMP-1 were more associated with cancer phenotype than other types of MMP and TIMP. The influence of human stromal tissues (peritoneum, myometrium, ovary) on the secretion of MMPs and TIMPs was examined by addition of these stromal tissues culture medium to human ovarian cancer cells (SAOV). All conditioned medium of stromal tissues could increase in both MMP-2, MMP-9, TIMP-1, and TIMP-2 activity in SAOV cells. Fraction (> 100kD) of conditioned medium of peritoneum could increase remarkably in MMP-9, and this incr

We evaluated the anaerobic threshold (AT) in 104 normal pregnant women mainly in their second trimester. Each exercise testing was performed by using incremental bicycle ergometry at a pedal frequency of 60rpm while monitoring maternal heart rate, maternal blood pressure, minute ventilation (VE),oxygen uptake (VO2) and carbon dioxide output (VCO2) every 30 seconds. The AT was determined by estimating the point of departure from linearity of the plot of VE as a function of VO2. We could determine the AT in 100 cases (96.2%), and their AT values and the heart rates at AT point were 14.7 +/- 1.7ml/min/kg (mean +/- S.D.) and 128 +/- 12bpm, respectively. AT decreased significantly (p < 0.01) with the length of gestation, because of increased maternal body weight. The heart rate at the AT also declined significantly (p < 0.05) with gestational age, but it had no relation with maternal body weight. These results show that the maternal heart rate becomes hard to increase with gestational age, so that the maximal heart rate, as an index of exercise intensity during exercise should decrease with advancing gestational age.

We performed a systematic retroperitoneal lymph node dissection (RPLND) on 137 patients with primary ovarian carcinoma, of whom 97 had undergone RPLND during the primary surgery before chemotherapy and 40 had undergone RPLND during the secondary cytoreductive surgery after preoperative chemotherapy. The tentative staging of the ovarian carcinoma used in this study was determined according to the FIGO criteria without considering the pathologic findings of retroperitoneal lymph nodes. Nodal metastasis was seen in 21.9% (30/137) of them. Thirteen had positive pelvic lymph nodes (PLN) but no positive para-aortic nodes (PAN). Eleven had both positive PLN and positive PAN. Six had positive PAN but no positive PLN. The PAN was the most frequent site of metastasis (17/137). Next were the common iliac, obturator, and lateral group of deep inguinal nodes. Solitary metastasis in the patients who had undergone RPLND during the primary surgery was seen in a PAN and a common iliac node. Among 24 patients with PLN metastasis, there was a significant (p < 0.05) difference in the number of positive PLN between the patients with PAN metastasis (5.27 +/- 3.00) and the patients without PAN metastasis (2.62 +/- 1.66). These results indicate that the PAN and common iliac nodes are the most important site of nodal metastasis in ovarian carcinoma. The metastasis to PLN such as obturator node and internal iliac node seems to occur independently of the PAN metastasis, and the PAN metastasis occurs not only through the direct route but also as a consequence of extension of PLN metastases. Systematic retroperitoneal lymph node exploration therefore seems to be necessary to clarify the lymph node status.

Forty-eight cases of human uterine cervical cancer were examined for the expression of c-myc protein by immunohistochemical staining. The overexpression of c-myc was detected in 17 of 48 cases (35%), which is consistent with previous reports. The frequency of c-myc overexpression was not associated with the clinical stage. Relapse was observed in 7 of 15 cases (47%) which had overexpression of c-myc (mean follow-up period: 35 months), whereas relapse was observed in only 3 of 30 cases (10%) which did not overexpress c-myc (mean follow-up period: 33 months). The five-year survival rate was significantly lower in the cases overexpressing c-myc than in those not overexpressing it. This indicates that the overexpression of c-myc may be associated with a high risk of relapse and poor prognosis. We also analysed the correlation between lymph node metastasis, cervical stromal invasion and c-myc overexpression, and did not find any correlation between them. These results suggest that the overexpression of c-myc in cervical cancer may be a prognostic indicator for predictive testing.