Yasuhiro Udagawa

Differences in lectin binding patterns of normal human endometrium between proliferative and secretory phases

SummaryLectin binding patterns in normal human endometrium were examined by light and electron microscopy using seven different lectins (ConA, WGA, RCA, PNA, UEA-1, DBA, and SBA). For light microscopic observations, criteria based on the incidence and intensity of cells positive for the lectin staining were adopted to evaluate the different staining patterns of the proliferative and secretory endometria obtained by the avidin-biotin-peroxidase complex (ABC) technique. At the light microscopic level, ConA, WGA, and RCA stained endometrial glandular cells in both phases. The number of PNA-positive cells with the binding sites entirely limited to the apical surface tended to be reduced slightly in the secretory phase. UEA-1 weakly stained the apical surface of glandular cells in the proliferative phase but not in the secretory phase. Among the lectins used in this study, DBA and SBA displayed remarkable changes between the phases. That is, in the proliferative phase they produced only a faint or slight positive stain at the apical surface, but the incidence and intensity of DBA- and SBA-positive glandular cells increased in the secretory phase. By electron microscopy, the reaction product of ConA was observed in the plasma membrane, endplasmic reticulum, nuclear envelope, and the Golgi apparatus, and the binding sites of RCA and DBA were observed in the plasma and, Golgi membranes. Between both phases, the reactivity of ConA and RCA showed almost no change. However, the secretory endometrial cells containing the DBA-positive Golgi apparatus were markedly increased in number compared with the proliferative ones bearing the lectin-positive organelles. These results indicate that DBA and SBA are useful for detection of hormone-mediated changes of normal human endometrium, and suggest that such changes are closely related to the Golgi apparatus.

Serum placental alkaline phosphatase (PLAP) in gynecologic malignancies — with special reference to the combination of PLAP and CA54/61 assay

Serum placental alkaline phosphatase (PLAP) levels in patients with gynecological tumors were measured by two kinds of enzyme-antigen immunoassay kits provided by Innogenetics (PLAP-I) and Sangtec Medical (PLAP-S), and the combination assays for PLAP with other tumor markers were studied. None of the healthy women studied were PLAP positive, and the positive rate in patients with benign ovarian tumors was less than 6%. The positive rate in patients with ovarian cancers was about 35%, which was higher than the rates for other cancers. It was significantly higher in patients with ovarian serous cystadenocarcinoma (60%). Remarkably high PLAP-I values were observed in patients with dysgerminoma. By the combination assay for PLAP with CA54/61 antigen in ovarian cancers, the diagnostic efficiency increased compared with that for PLAP and CA125. We conclude that PLAP is useful in the diagnosis of ovarian serous cystadenocarcinoma and dysgerminoma and that CA54/61 is the better partner for the combination assay.

High Expression of Uridine Diphosphate‐galactose: Lc3Cer βl‐3 Galactosyltransferase in Human Uterine Endometrial Cancer‐derived Cells as Measured by Enzyme‐linked Immunosorbent Assay and Thin‐layer Chromatography‐immunostaining

We have developed a new procedure for the selective determination of β1‐3 and β1‐4 galactosyltrans‐ferases with Lc3Cer as the substrate and the microsomes of fetal and adult porcine livers as the enzyme sources. This method was based on the detection of such products as Lc4Cer for β1‐3 galactosyltransferase (β1‐3GT) and nLc4Cer for β1‐4 galactosyltransferase (β1‐4GT), with monoclonal anti‐Lc4Cer and anti‐nLc4Cer antibodies, respectively. This method thus enabled us to differentiate the activity of β1‐3GT from that of 4bT1‐4GT with a high degree of sensitivity. The method was then used to determine the activities of both enzymes in human gynecological carcinoma‐derived cells. Four of the five cell lines derived from uterine endometrial cancer expressed significantly high levels of specific activity of β1‐3GT among the cell lines examined, while their β1‐4GT activities were less than 20% of that for β1‐3GT in the endometrial carcinoma‐derived cells. On the other hand, a higher specific activity of β1‐4GT than that of β1‐3GT was detected in the cell lines derived from uterine cervical and ovarian cancers. These findings were thus found to correlate closely with the rate of expression of Lc4Cer‐ and nLc4Cer‐based carbohydrate chains in the cell lines based on the results of immunohistochemical staining.

Phenotypic alterations of a carbohydrate antigen, blood group A type 3 chain, in neoplastic transformation of uterine cervix

A monoclonal antibody, MRG-1, was established by use of a human ovarian mucinous cyst-adenocarcinoma-derived cell line, RMUG-L, as immunogen. Following its establishment, biochemical analysis revealed that its epitope was blood group A type 3 chain. Using MRG-1 as an immunohistochemical probe, uterine cervical neoplastic lesions including dysplasia, carcinomain situ, and invasive carcinoma were investigated. Light-microscopically, normal squamous epithelium showed a strong positive reaction along the cell surface region exclusively in the intermediate cell layer. On the other hand, intracellular structures were very often strongly stained in squamous cell carcinoma. Under the electron microscope, MRG-1 binding sites in squamous cell carcinoma cells were found to be in intracytoplasmic vesicular structures as well as in the plasma membrane. This marked difference in the antigen distribution was found to be a phenomenon associated with cervical neoplastic transformation.