Serum soluble ICAM-1 levels in the patients with cervical cancer.

BACKGROUND: We hypothesize that serum levels of the soluble intercellular adhesion molecule-1 (s-ICAM-1) in patients with cervical cancer are significantly higher than that in healthy donors and this is examined in vivo, and in vitro. METHODS: We measured the serum levels of the s-ICAM-1 in patients with cervical cancer. Furthermore, in vitro, we examined the relationship between the expression of ICAM-1 on the cell surface of cultured cervical cancer cells and the s-ICAM-1 levels in the spent media. RESULTS: The mean+/-s.d. in the patients with cervical cancer 884.4+/-332.4 ng/ml were significantly (all, p<0.001) higher than those of normal controls (mean 364.6+/-134.8 ng/ml) and in patients with benign disease (536.3+/-204.8 ng/ml). Interferon-gamma (IFN-gamma) treated cells expressed more ICAM-1 on cell surfaces than did the non-treated cells, and s-ICAM-1 shed in the spent media from IFN-gamma treated cells were much higher than those of non-treated cells. CONCLUSIONS: These results indicate that the reason for increase of s-ICAM-1 in the patients with cervical cancer were both expression and shedding of ICAM-1 from cervical cancer cells especially influenced by cytokines, for example, IFN-gamma from surrounding lymphocytes.

Visual findings and histologic diagnosis of pelvic endometriosis under laparoscopy and laparotomy.

OBJECTIVE: To assess the diagnostic value of visual findings in the diagnosis of endometriosis from the histological point of view. STUDY DESIGN: 212 specimens from 107 patients with benign (74.8%) or malignant (25.2%) disease were obtained by biopsy or resection under laparoscopy (65 patients) or laparotomy (42 patients). Ages ranged from 19 to 62 (mean age 36.4). Visual findings were classified according to the criteria established by the Endometriosis Committee of the Japan Society of Obstetrics and Gynecology in 1993. Specimens were stained with hematoxylin-eosin, and 15 cases with periodic acid-Schiff stain or silver impregnation stain. RESULTS: Among pigmented lesions, endometriosis was found in 73.0% of specimens from the pelvic peritoneum and in 56.4% of those from the ovaries. Blueberry spots in the pelvic peritoneum as well as ovarian chocolate cysts showed the highest positive rate. In the presence of multiple or complex pigmented lesions of the pelvic area, the rate was still higher (88.6%). Those rates were due to our inclusion of inaccurate and incomplete biopsy specimens. Endometriosis of nonpigmented lesions was found in only 11 patients (12.0%) who also had pigmented lesions and/or adenomyosis. CONCLUSION: The laparoscopic diagnosis of endometriosis can be made only when multiple complex pigmented lesions are observed, but, otherwise, histopathological confirmation is necessary.

Endocrinological and histological changes after treatment of uterine leiomyomas with danazol or buserelin.

The following results were obtained from our investigations performed to compare the effects of danazol (400 mg/day orally for 4 to 6 months, n = 164) with those of buserelin (900 micrograms/day intranasally for 4 to 6 months, n = 83) on patients with uterine myomas. The effectiveness of buserelin (76.1%) in shrinking uterine myomas was better than that of danazol (56.7%). The serum-estradiol and progesterone levels were lower in the buserelin group than in the danazol group. The estrogen and progesterone receptors in a hysterectomied uterus were positively stained even at the end of those therapies. The degree and frequency of hyaline degeneration were higher in the buserelin group than in the danazol group. Thus, the shrinking of uterine myomas and the degree of hyaline degeneration might be caused by the lowering of uterine-tissue hormones, particularly of estradiol levels.

Macrophage activation factors and cytokines in peritoneal fluid from patients with endometriosis.

Endometriosis is often associated with infertility, which might be a consequence of intraperitoneal macrophage activation and various immunological events related to it. To determine what triggers macrophage activation and what immunologic conditions occur in the peritoneal cavity of a patient with endometriosis, we measured the amounts of phospholipase A2, cholesterol fractions (HDL, LDL, and VLDL), interleukins (IL)-1 and 6, granulocyte-macrophage-colony stimulating factor (GM-CSF), and phosphatidylserine in peritoneal fluid. Peritoneal-fluid samples were collected during laparoscopy or laparotomy from 14 patients with endometriosis and from 11 patients with other diseases (5 with myoma uteri, 5 with benign ovarian cysts and 1 with a double uterus) as controls. With regard to phospholipase A2, there were no significant differences between the endometriosis group and the control group. LDL was significantly lower in the endometriosis group than in the control (p < 0.05). The levels of IL-1 and GM-CSF were similar in both groups; IL-6, however, was higher in 7 endometriosis patients with associated infertility than in the 9 non-endometriosis fertile patients. These findings suggest that inflammatory tissues are not related to the activation of intraperitoneal macrophages in endometriosis patients. Cholesterol fractions (HDL or LDL) might participate in macrophage activation in these patients, but this remains to be clarified. The high IL-6 levels observed in infertile endometriosis suggest that some immunological events that prevent patients from becoming pregnant take place in women with endometriosis-associated infertility.

[A study of intercellular adhesion molecule-1 (ICAM-1) in cervical cancer].

Intercellular adhesion molecule-1(ICAM-1), one of the adhesion molecules, plays an important role in target-cell lysis by immune cells. In this study we examined the expression of ICAM-1 in 15 cases of cervical cancer. We also examined ICAM-1 expression on cervical cancer cell lines in response to cytokines. Immunohistochemistry revealed expression (4 positive among 15 cases) of ICAM-1 in cervical cancer. Furthermore, the expression of ICAM-1 on cervical cancer was correlated with the degree of mononuclear cell infiltration, bearing CD3. On the other hand, the expression of ICAM-1 was not correlated with Class I or Class II antigens in cervical cancer. The expression of ICAM-1 on cervical cancer cell lines was augmented by in vitro treatment with interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha. These results suggest that the expression of ICAM-1 on cervical cancer might be modified by cytokines, and ICAM-1 expression on cervical cancer might augment the host immune reaction. So it will be possible to use cytokines in immunotherapy.

[In vitro study on the effect of sex steroid and growth factor on growth and laminin, collagen IV, and tissue plasminogen activator production of normal endometrial cells and endometrial cancer cells in culture].

The mechanism of the invasion and proliferation of endometrial cancer is closely related to interactions between the endometrial glands and stroma. In this study, we examined the biological role of sex steroids (estradiol; E2, progesterone; P) and growth factors (epidermal growth factor; EGF, transforming growth factor-beta; TGF-beta) on cell growth and laminin, collagen IV and tissue plasminogen activator (t-PA) production of normal endometrial cells and endometrial cancer cells in culture. Normal endometrial gland cells and stromal cells, and endometrial cancer cell lines (Ishikawa, OMC-2) were used. E2, P, EGF and TGF-beta were added to the culture in physiological concentrations. The growth of normal endometrial gland cells was promoted by E2 and EGF, whereas that of Ishikawa cells and OMC-2 cells was promoted by EGF. E2 enhanced the effects of EGF in normal endometrial gland cells. The growth of normal endometrial stromal cells was not affected by them. OMC-2 was inhibited by anti-EGF receptor antibody. On the other hand, the production of laminin and collagen IV of these cultured cells was inhibited by EGF and promoted by TGF-beta, whereas that of t-PA was promoted by EGF and inhibited by TGF-beta. These results suggest that the growth of normal endometrial gland cells with estrogen receptor (ER) is controlled by both E2 and EGF, whereas that of endometrial cancer cells is affected only by EGF, and those cells without ER depend particularly on the autocrine growth mechanism of EGF.(ABSTRACT TRUNCATED AT 250 WORDS)

[Study on the management of dysplasia of the uterine cervix].

In this study, we investigated problems on the management of dysplasia on the basis of our clinical data obtained in the past ten years. In addition, we also examined the biological significance of protooncogene expression and human papilloma virus (HPV) infection in the initiation and promotion of dysplasia. Among 540 cases of dysplasia we have managed, 48.8% of the cases of mild, moderate and severe dysplasia which we followed up for more than 6 months regressed, and 24.1% of the cases progressed. The cure rate for laser therapy based on the follow up for 6 to 96 months was significantly high (97.9%) in 342 cases treated by the cone method, and low (89.5%) in 38 cases treated by the vaporization method. Preoperative histological findings confirmed 60.5% of postoperative findings, and several early cervical carcinomas were found in preoperative cases of dysplasia by laser conization. Among 28 cases (8.2%) of incomplete excision, 24 cases (85.7%) regressed spontaneously. On the other hand, the positive rate of immunostaining of epidermal growth factor receptor (EGF-R) and c-myc oncogene product (c-myc protein) increased from mild or moderate to severe dysplasia, and the positive rate for EGF-R was significantly high (80.0%) in the progressive group. The positive rate for HPV genome was quite low (16.0%) in dysplasia. Among them, EGF-R was most associated with the prognosis of dysplasia. These results suggest that laser conization should be performed for many cases of dysplasia because of the preoperative possibility of the existence of early cancer, and EGF-R is also useful in determining the necessity for therapy for dysplasia.

[In vitro studies on the expression of c-myc oncogene product in gynecological cultured cancer cells].

In this study, immunocytochemical and biochemical detection of c-myc protein in gynecological cultured cancer cells were performed together with gene expression of the cells. OMC-1 (cervical squamous carcinoma cell line), OMC-2 (endometrial adenocarcinoma cell line), OMC-3 (ovarian mucinous adenocarcinoma cell line) and OMC-4 (cervical adenocarcinoma cell line) were used. Immunocytochemically, c-myc protein was detected in both nuclei and cytoplasms of cultured cells when they were fixed in 95% ethanol, 10% formalin and 4% paraformaldehyde (PFA) including 10mM NaCl. However, it was detected in the nuclei of almost all of the cells in nuclei of the cells when they were fixed in 4% PFA including 1,000mM NaCl. Western blotting against a nuclear fraction of the cells demonstrated 66Kd protein in OMC-1 and 62Kd protein in OMC-2,3,4, respectively. They were completely absorbed by c-myc synthetic peptide. However, there was no reaction against the cytoplasmic fraction of the cells. Slot blot hybridization against the DNA of the cells demonstrated 15 times and 5 times c-myc gene amplification in OMC-2 and OMC-4, respectively. These results suggest that OMC-1,2,3,4 can be used as positive controls for the immunocyto- and histochemical detection of c-myc protein in clinical materials. However, it must be noted that the redistribution of c-myc nuclear protein into the cytoplasm may occur in the fixation process.

[A study of the expression of class II antigen on uterine cervical cancer cells and cell lines].

An immunohistochemical method with monoclonal antibody to DR antigen was used to study class II antigens and HLA-DR antigen in 20 patients with uterine cervical cancer. Eight patients had various amounts of DR antigen. Regional infiltrating lymphocytes were also examined with anti-Leu 1, Leu 2a, Leu 3a, and Leu 10. Among the many infiltrating T cells, Leu 3a-positive cells were relatively predominant surrounding the cancer nests which contained DR antigen. Cervical cancer cell lines OMC-1 and OMC-4 both demonstrated DR antigen. Interferon (IFN)-gamma treatment enhanced the expression of DR antigen in OMC-1 and OMC-4. The DR antigen in OMC-1 and OMC-4 were capable of stimulating allogenic lymphocytes in mixed lymphocyte reaction (MLR), and their stimulatory activity was significantly enhanced by IFN-gamma treatment. These results demonstrated the expression of class II antigen, especially DR, on some cervical cancer cells and cell lines and showed that the DR antigen in uterine cancer cell lines can stimulate MLR.

[EGF receptor and effects of EGF on growth and tumor marker secretion in uterine cervical cancer cells].

Effects of EGF on proliferation and tumor marker secretion of cervical cancer cells are reported together with the characteristics of EGF receptors on the cells. TA-4 producing cell line (OMC-1) originating from cervical squamous cell carcinoma and CA-125 producing cell line (OMC-4) originating from cervical adenocarcinoma, were used. Scatchard plot of EGF binding to OMC-1 indicated a single class of binding sites with a dissociation constant (Kd) of 360pM, whereas that of OMC-4 was curvilinear suggesting two classes of binding sites with a Kd of 170pM and 510pM. The theoretical maximum number of binding sites of OMC-1 and OMC-4 was 2.4 X 10(4) and 1.6 X 10(5), respectively. Effects of EGF on growth were studied by monitoring cell number and the incorporation of 3H-thymidine into the DNA of the cells. OMC-1 was stimulated by EGF at low concentrations (0.01-0.1nM) and inhibited at higher concentrations. OMC-4 was not stimulated by EGF. The TA-4 secretion of OMC-1 was slightly stimulated by EGF at low concentrations (0.01-1nM) and significantly stimulated at high concentration (10nM). The CA-125 secretion of OMC-4 was not stimulated by EGF. These results suggest that there are some differences between cervical squamous cell carcinoma and adenocarcinoma in the mechanisms of regulation of proliferation and tumor marker secretion by EGF.

[Immunohistochemical studies on epidermal growth factor receptor in experimental squamous cell carcinoma of the uterine cervix of mice].

The tissue localization of epidermal growth factor receptor (EGF-R) in experimental squamous cell carcinoma of the mouse uterine cervix was examined immunohistochemically. Carcinoma was induced by the insertion of a 20-methylcholanthrene (MC)-impregnated thread into the cervical canal of the mouse. Tissue sections (of normal columnar epithelium, proliferation, atypical proliferation, early invasive carcinoma, invasive carcinoma and metastatic carcinoma) were stained by the avidin/biotin immunoperoxidase technique using anti-EGF-R monoclonal antibody. Normal columnar epithelium was negative for EGF-R, whereas proliferation was partly positive. The lesions of atypical proliferation and early invasive carcinoma had a positive staining for EGF-R. The staining for EGF-R declined in the lesion of invasive carcinoma. Metastatic carcinoma was not stained for EGF-R. These results suggest that EGF-R may play an important role in the early stage of carcinogenesis of the mouse uterine cervix induced by 20-MC.