Adjuvant chemotherapy in stage I uterine endometrial carcinoma.

OBJECTIVE: We have assessed prognostic factors and the efficacy of adjuvant chemotherapy in stage I uterine endometrial carcinoma. METHODS: 251 primary surgically treated stage I patients were studied. Prognostic factors were evaluated and 5-year and 10-year survival rates were compared in patients with lymph-vascular space invasion to investigate whether adjuvant chemotherapy improves survival. RESULTS: The overall 5-year and 10-year survival rates were 94% and 93%. Multivariate analysis indicates that lymph-vascular space invasion is the most significant prognostic factor in both 5- and 10-year survival rates (P<0.001 at both times) and stage/depth of invasion is significant for the 10-year survival rate (P=0.04). Of 54 patients with lymph-vascular space invasion, statistically significant differences were observed in 10-year survival rate (P=0.02) between patients who had surgery followed by adjuvant chemotherapy (n=23) and patients who had surgery alone (n=31). Toxicities were mild to moderate (30%). CONCLUSIONS: The clinical importance of lymph-vascular space invasion and the efficacy of adjuvant chemotherapy were confirmed. This observation warrants a larger comparative study with adjuvant chemotherapy.

Expression of vascular endothelial growth factors (VEGF-A/VEGF-1 and VEGF-C/VEGF-2) in postmenopausal uterine endometrial carcinoma.

OBJECTIVE: We investigated the expression of two angiogenic vascular endothelial growth factors, VEGF-A/VEGF-1 and VEGF-C/VEGF-2, in 228 cases of uterine endometrial carcinomas from postmenopausal patients to evaluate the correlation with histopathologic features and clinical outcome. METHODS: Immunohistochemistry was used to assess VEGF-A/VEGF-1 and VEGF-C/VEGF-2 expression in 228 primary surgically treated cases of postmenopausal endometrial carcinomas and the results were statistically analyzed in relation to vascular invasion, depth of invasion (myometrial vs serosal-parametrial invasion), lymphatic vessel invasion, lymph node metastasis, disease-free 5-year survival rate (DF5YR), and disease-free 10 year-survival rate (DF10YR). RESULTS: The results of univariate analysis showed that VEGF-A/VEGF-1 and VEGF-C/VEGF-2 expression correlated with vascular invasion (P < 0.0001, P = 0.0006), depth of invasion (P = 0.0004, P = 0.043), lymphatic vessel invasion (P = 0.021, P < 0.0001), lymph node metastasis (P = 0.0017, P = 0.0008), DF5YR (P = 0.0081, P = 0.0002), and DF10YR (P = 0.0077, P = 0.0001). Multivariate analysis showed that lymph node metastasis (P = 0.0017, P = 0.0008), parametrial-serosal invasion (P < 0.0001, P < 0.0001), and VEGF-C/VEGF-2-positive status (P = 0.03, P = 0.01) were significant factors in DF5YR and DF10YR. CONCLUSIONS: We conclude that VEGF-A/VEGF-1 and VEGF-C/VEGF-2 expression was predictive of these histopathologic features of endometrial carcinoma and clinical outcome.

Prognostic factors relating to survival in uterine endometrioid carcinoma.

OBJECTIVE: Epidemiologic and clinicohistopathologic prognostic factors of uterine endometrioid carcinomas were analyzed. The association of estrogen related factors, focused on adenomyosis in the prognosis of endometrioid carcinomas was also examined. METHODS: Risk factors of surgically treated 286 patients with endometrioid carcinoma (Stage I-III) were statistically analyzed. RESULTS: Overall a recurrence-free 5-year survival rate was 81% (Stage I, 94%, Stage II, 71% and Stage III, 40%). Significant prognostic factors were lymph node metastases (P = 0.0035) and serosal/parametrial invasion (P = 0.014) by multivariate analysis. Endometrioid carcinomas with co-existing adenomyosis tend to be associated with endometrial hyperplasia (P = 0.04, Fisher's exact test), diagnosed in less invasive status (myometrial invasion, P = 0.004 and serosal/parametrial invasion, P = 0.006) and therefore have a favorable prognosis (P = 0.01, log rank test). CONCLUSIONS: A favorable prognosis of endometrioid carcinomas with co-existing estrogen related factors (adenomyosis and endometrial hyperplasia) was suggested.

[Clinical outcome of an anticancer agent screening test with primary culture cells].

Cancer cells are tested for sensitivity to anticancer agents before the drugs are used for treatment in clinical settings, but the results of these tests have not always been reported fully. Since 1973, one of the authors (Hirono) has been performing sensitivity tests of anticancer drugs by using primary culture cells derived from human cancer tissue, taking into account in vivo status and taking great care to reduce the number of procedural errors. From 1991, flow cytometry has also been adopted for use in anticancer drug sensitivity tests. In the present study, the outcomes of cancer patients were compared by dividing them into a group treated only with anticancer agents to which cancer cells had responded in sensitivity tests, and a group treated with other drugs. The subjects consisted of 132 patients with endometrial cancer stage III (n = 26) and IV (n = 10), ovarian cancer stage III (n = 26) and IV (n = 7), peritonitis carcinomatosa (n = 18), and suspected advanced ovarian cancer (n = 45). There was a significant prolongation in median survival time and survival time among the non-survivors according to the results of a Kaplan-Meier analysis. These findings suggest that sensitivity testing of cancer cells to anticancer drugs should be performed before the start of cancer chemotherapy in the clinical setting.

Papillary carcinoma in a huge intrathoracic goiter with tracheal stenosis and invasion. Case report.

A rare case in tracheal invasion by a papillary carcinoma in an intrathoracic goiter is reported. Subtotal thyroidectomy and tracheoplasty were performed. Surgery should be considered as first-line treatment of intrathoracic goiter in order to avoid future compression of the respiratory tract and to remove possible concomitant malignant lesion.

[Result of individualized chemotherapy with a newly developed in vitro chemosensitivity testing in ovarian cancer].

By means of a newly developed in vitro chemosensitivity test based on the morphological changes in the nucleus (nuclear damage assay) as previously described, we were able to screen currently available anticancer drugs within 24 hr with a 100% success rate. The nuclear damage assay was used to determine the chemosensitivity in 50 patients (66 assays) with ovarian cancer. The response rate for the 13 patients with measurable tumors, 8 of whom showed resistance to CAP (cyclophosphamide, adriamycin, and cisplatin) therapy, was 46 percent when they were given various combination chemotherapy protocols consisting of more than one active agent selected from group A and B agents by the nuclear damage assay. The newly developed in vitro chemosensitivity test proved to be useful when selecting a second line combination for patients with CAP-resistant ovarian cancer.

A newly developed in vitro chemosensitivity test (nuclear damage assay): application to ovarian cancer.

With a newly developed in vitro chemosensitivity test based on the morphological changes of nuclear damage (nuclear damage assay) described here, we were able to screen currently available anticancer drugs within 24 hr with a 100% success rate. In preclinical chemotherapy using four human ovarian epithelial tumor cell lines and their xenografts in nude mice, the in vitro/in vivo response (sensitive/sensitive and resistant/resistant) rate was 94%. The nuclear damage assay was used to determine the chemosensitivity in 49 patients (60 assays) with ovarian cancer. The response rate of the 13 patients with measurable tumors, 9 of whom showed resistance to CAP (cyclophosphamide, adriamycin, and cisplatin) therapy, was 46% when the patients were given various combination chemotherapy protocols consisting of more than one active agent selected from group A and B agents by the nuclear damage assay. The newly developed in vitro chemosensitivity test proved to be useful when selecting a second-line combination chemotherapy for patients with CAP-resistant ovarian cancer.

Mechanisms of resistance to cis-diamminedichloroplatinum (II) in a rat ovarian carcinoma cell line.

A cis-diamminedichloroplatinum (II) (cisplatin)-resistant subline (Cis-Ptr) demonstrated 20-fold greater resistance to the cytotoxic effects of cisplatin, compared with the parental cloned rat ovarian carcinoma cell line (ROT 68/C1). The uptake of cisplatin into the Cis-Ptr cells was identical to that into the ROT68/C1 cells in vitro and in vivo. Glutathione activity in a cytoplasmic extract was 1.4-fold and 1.8-fold greater in the Cis-Ptr cells than in the ROT68/C1 cells in vitro and in vivo, respectively. There was no difference between the ROT68/C1 and Cis-PTr cells in 195m cisplatin binding per micrograms DNA. DNA repair of cisplatin DNA damage was increased in the Cis-PTr cells but not in the ROT68/C1 cells. These results suggest that the mechanisms of resistance to cisplatin in rat ovarian carcinoma cells involve increased activity of the DNA repair system and increased cytosolic binding to thiols may also be involved.

The isolation and characterization of cisplatin-resistant rat ovarian cancer cells.

A rat ovarian cancer cell subline (Cis-Ptr), which became approximately 20-fold resistant to cisplatin, was developed after continuous exposure of the cisplatin-sensitive parent cell line (ROT68/C1) to increasing doses of the drug in vitro. Both the ROT68/C1 and Cis-Ptr cells were tumorigenic in isologous (Sprague-Dawley strain) newborn rats and only the tumors developed by inoculation of Cis-Ptr cells showed resistance to cisplatin in vivo. Resistance towards cisplatin was accompanied by phenotypic changes of the undifferentiated adenocarcinoma cells including enlargement of the cell and nucleus and a slower growth rate both in vitro and in vivo. Compared to the ROT68/C1 cells, the Cis-Ptr showed an early recovery of DNA synthetic activity after exposure to cisplatin. Cross-resistance of the Cis-Ptr cells was found only to a cisplatin analog, carboplatin. These results suggest that our cisplatin resistant rat ovarian cancer cells are useful in the investigation of the characteristics and mechanisms of cisplatin resistance.

[Fundamental studies on tumor angiogenesis factors with a choriocarcinoma model].

We have studied biologically active substances, tumor angiogenesis factors (TAF), which are supposed to be associated with the proliferation and metastases of choriocarcinoma cells. Eight human choriocarcinoma cell lines were used in the present study. TAF activity was assayed by bioassays using BALB/c mice subcutaneous tissue, chicken chorioallantoic membrane (CAM) and rabbit cornea in vivo and by proliferation of endothelial cells in vitro. 1. We found a positive correlation between the size of tumors developed in xenotransplantation and the number of blood vessels in the tumor tissues. A correlation between the number of blood vessels and TAF activity was also found. 2. With a gel-filtration method, TAF activity was observed in the common fractions in every cell line. The molecular weight of TAF was more than 10,000 daltons. 3. There was a heterogeneity of TAF activity among the cell lines. From these results, it appears that TAF initiates the choriocarcinoma cell-proliferation.

Sensitivity of human germ-cell-tumor cell lines to human interferons.

We examined the sensitivity of four human germ-cell-tumor cell lines exhibiting different stages of differentiation to human interferons (IFNs) in vitro. The cell lines were derived from two embryonal carcinomas (NEC 8 and NEC 14), a choriocarcinoma (IMa), and a yolk-sac tumor (HUOT). Treatment with poly I:C induced IFN production in IMa and HUOT cells, but not in NEC-8 and NEC-14 cells. In the two embryonal-carcinoma cell lines, the addition of IFN-alpha, IFN-beta, and IFN-gamma did not prevent infection by vesicular stomatitis virus and encephalomyocarditis virus. Also, in these two lines, 2-5A synthetase was not induced by the addition of IFN-alpha. In contrast, both IMa and HUOT showed sensitivity to the antiviral action of IFN-alpha and IFN-beta against the two viruses, and 2-5A synthetase was induced by IFN-alpha. IFNs added at doses of up to 1000 IU/ml had no antiproliferative effect on NEC 8, NEC 14, and HUOT, whereas colony formation by IMa cells was greatly suppressed by all three forms of IFN. These results indicate that the production of and sensitivity to IFN are developmentally regulated and are related to the level of differentiation of human germ-cell stem cells.

Tumor angiogenesis activity of human choriocarcinoma cells grown in vitro.

The growth of five different kinds of human choriocarcinoma cell lines in vitro was quite alike. However, two distinct types of rapid and slow growth were observed in tumors grown in the hamster cheek pouch. The rapidly grown tumor tissues (BeWo, JEG-3, and NUC-1) involved significantly large numbers of blood vessels per microscopic field, as compared to slowly grown ones (SCH and HM). The vascular response was assayed using cell-free crude tumor angiogenesis factors (TAF) on chorioallantoic membrane (CAM) of the chick embryo. In all cell lines TAF activity correlated well with the extent of tumor growth in vivo. Gel filtration analysis showed that relatively high-molecular-weight (more than 10,000) factors could induce neovascularization in the both assays using CAM and rabbit cornea. These results suggest that a heterogeneity of TAF activity is present among human choriocarcinoma cell lines and tumor growth in xenograft depends on the secretion of TAF by the tumor cells themselves.

The presence of extraganglionic fluorescent neurons in the myenteric plexus of the guinea pig small intestine.

Fluorescent histochemical observations of the small intestine of the guinea pig demonstrated that single fluorescent cell bodies, separate from the ganglia, were present in the myenteric plexus. These cell bodies gave rise to single processes which entered the ganglia or the interganglionic strands. They were of a very small size, and the intensity of their fluorescence increased by pretreatment with L-DOPA and nialamide. Interruption of extrinsic nerve pathways to the small intestine caused a disappearance of the meshwork of fluorescent fibers in the myenteric plexus; but in some areas a fluorescent fiber which supplied its terminal to the ganglion was seen to remain. A photograph taken from the denervated myenteric plexus revealed that a long process arising from an extraganglionic cell entered the ganglion and ramified into terminal branches.

Invasion potential of human choriocarcinoma cell lines and the role of lytic enzymes.

The chick chorioallantoic membrane (CAM) was used as an assay system to examine the invasive potential of human choriocarcinoma cell lines. When 5 X 10(6) cells were inoculated into the CAM at the 10th day of postfertilization, three of eight cell lines formed extensively invasive tumors within the CAM. A tendency to correlation between the tumorigenic potential of cell lines in hamster cheek pouches and their invasive potential in the CAM was noted. In addition, the invasive capacity of cell lines correlated well with the amount of collagenase but did not correlate with the amount of plasminogen activator or cathepsin B secreted by them. It is concluded that a heterogeneity of invasive potential in the CAM exists among human choriocarcinoma cell lines and the role of the collagenase secreted by them is suggested.

Localization of carbonic anhydrase in the rat lung.

The localization of carbonic anhydrase in the rat lung has been demonstrated, at light and electron microscopic levels, by the cobalt bicarbonate histochemical method of Hansson. Focal deposits of the cobalt sulfide reaction product were found not only in the capillary endothelium of the alveolar walls, but also in the small and large alveolar cells. The histochemical reaction was abolished by two potent inhibitors, acetazolamide (10(-5) to 10(-6) M) and KCNO (5 x 10(-3) to 10 x 10(-3) M). Physiological assay with Maren's method indicated that values for carbonic anhydrase activity in rat lung are 4.4 +/- 0.8 UA/mg of protein, 25.0 +/- 5.5 UA/mg of nitrogen, and 369 +/- 86 UA/g of wet weight. In addition, it was calculated that after fixation in glutaraldehyde-formaldehyde-picric acid about 9% activity is retained.