Expression of blood group-related antigens in neoplastic uterine cervix

Expression of blood group antigens in normal, displastic and tumoral uterine cervix from 35 hysterectomised women with carcinoma of the cervix was investigated; the results were correlated with patients' ABH phenotype and secretor status. We used an indirect immunoperoxidase technique and a panel of monoclonal antibodies and lectins directed against different antigenic specificities. Anomalous expression of blood group antigens in premalignant lesions from cervix was found. Partial loos of expression of blood group antigens and some lectins in different grades of cervical intraepithelial neoplasia, and a total loss of expression in CIN III and in infiltrating carcinoma of the cervix from secretor patients was revealed. The findings herein described confirm the importance of these antigens as tumour markers and they might be useful for the study of cervical carcinogenesis.     

Blood group-related antigens in human germ cell tumors

With the use of immunohistochemical techniques, seven mouse monoclonal antibodies and the lectin from Ulex europaeus, detecting blood group antigens of the ABH and Lewis systems, have been used to define the distribution of these antigenic structures in germ cell tumors. The reagents used recognize the following blood group antigens: A, B, H, Lewisa, Lewisb, X (Lewisx), Y (Lewisy), and type I precursor antigen. Tumors from 29 patients were studied. Tumors studied consisted of pure embryonal carcinoma for eight patients, pure yolk sac tumor for two patients, embryonal carcinoma plus yolk sac tumor in one patient, and yolk sac tumor plus seminoma in one patient. Also studied were nine classic seminomas and a group of six patients with tumors classified as seminomas that exhibited atypical histological features. One patient had an anaplastic carcinoma arising from the mediastinum which could not be conclusively identified as a germ cell tumor morphologically and was analyzed separately. All embryonal carcinomas and yolk sac tumors exhibited strong positivity for type I precursor structure as detected by the K-21 monoclonal antibody. In marked contrast, there was non staining in classic seminomas but heterogeneous staining in five of six atypical seminomas. The majority of embryonal carcinomas and all yolk sac tumors studied demonstrated strong positivity for blood group antigen H. For seminoma, however, only one of the atypical cases and two of the classic cases (occasional cells) stained for H. Focal expression of the Y antigen was identified in 5 of 17 seminomas and in the majority of embryonal carcinomas and yolk sac tumors. Two yolk sac tumors and two classic seminomas expressed blood group X. The remaining blood group antigens were not expressed by seminomas while they were variably expressed by embryonal carcinoma and yolk sac tumors. These data suggest that K-21 and blood group antigen H may be distinguishing markers of nonseminomatous germ cell tumor versus seminoma. If so, it is possible that the heterogeneous expression of blood group substances in seminomas with atypical histologies is an indication of differentiation towards nonseminomatous germ cell tumor.     

Abnormal expression of blood group-related antigens in uterine endometrial cancers

The expression of A, B, and H group antigens, Lewis group antigens (Lewis(a), Lewis(b), Lewis(x), and Lewis(y)), and Lc4 and nLc4 antigens, the precursor antigens of both groups, was examined immunohistochemically with monoclonal antibodies in 9 normal endometria, 6 endometrial hyperplasias, and 31 endometrial cancers. 1) A, B and/or H antigens were detected in endometrial cancers at an incidence of 51.6%, while no distinct localization of these antigens was observed in normal endometria. H antigen, the precursor of A and B antigens, was particularly frequently detected in endometrial cancers. 2) An increased rate of expression of Lewis group antigens, particularly Lewis(b) antigen, was observed in endometrial cancers compared with its expression in normal endometria. 3) Lc4 and nLc4 antigens were detected in endometrial cancers at rates of 41.9% and 38.7%, respectively, these expressions being increased compared with those in normal endometria. 4) These results suggest that a highly abnormal expression of blood group-related antigens in endometrial cancers occurs not only at the level of A, B, and H antigens and Lewis group antigens, but also at the level of their precursor Lc4 and nLc4 antigens. 5) Lewis(a), Lewis(b), and Lc4 antigens, built on the type-1 chain, are more specific to endometrial cancers than their respective positional isomers, Lewis(x), Lewis(y), and nLc4 antigens, built on the type-2 chain.     

Immunohistological observation of blood group-related antigens in lung adenocarcinomas using monoclonal antibodies

The immunohistological distribution of blood group (BG)-related antigens including A, B, H type 2, and sialylated Lex in lung adenocarcinomas was examined using monoclonal antibodies. BG-A, B, and H type 2 compatible with the ABO status in tumor cells were expressed in 60% of the cases. Accumulation of H type 2, associated with loss of BG-A and B, was observed in tumor cells of patients with BG status other than 0. Tumor-associated antigens, Lex and sialylated Lex were detected in 36.0% and 72.0%, respectively. Modification of carbohydrate antigens in cancer may be associated with incomplete synthesis; accumulation of precursor antigen; and activated sialylation.     

Expression of LeY and extended LeY blood group-related antigens in human malignant, premalignant, and nonmalignant colonic tissues

The LeY determinant, a difucosylated type 2 blood group-related antigen, is a positional isomer of the Leb blood group antigen and a fucosylated derivative of the LeX antigen. The LeX antigen behaves like an oncodevelopmental tumor-associated antigen in human colon cancer, and extended polyfucosyl LeX antigens are more specific for colon cancer tissues than are simple, monofucosyl LeX antigens. The present investigation compared the expression of simple and extended LeY antigens in a variety of malignant and nonmalignant human colonic tissues to gain insight into the normal distribution and cancer-associated expression of these antigens. Monoclonal antibody AH-6, which recognizes the LeY epitope irrespective of its carrier carbohydrate chain, stained the majority of specimens regardless of malignant potential or location within the colon. In contrast, CC-1 and CC-2 monoclonal antibodies, which recognize extended LeY structures, and KH-1, which is specific to trifucosyl LeY, preferentially stained malignant colonic tissues and rarely stained normal colonic mucosae. Mucosa immediately adjacent to cancer usually stained with AH-6 but not with KH-1, CC-1, or CC-2. Extended or trifucosyl LeY antigen expression was limited exclusively to premalignant (adenomatous) polyps and was invariably absent from nonpremalignant (hyperplastic) polyps. Moreover, among adenomatous polyps, extended LeY antigen expression tended to correlate with three parameters of malignant potential: larger polyp size; villous histology, and severe dysplasia. AH-6 failed to distinguish between hyperplastic and adenomatous polyps. In second-trimester fetal colonic mucosa, AH-6 bound to both proximal and distal segments whereas KH-1, CC-1, and CC-2 bound only to proximal segments. We conclude that in human colon, the LeY hapten is an oncodevelopmental cancer-associated antigen and extended LeY antigens are highly specific markers for malignancy and premalignancy.     

Structural variations of blood group A antigens in human normal colon and carcinomas

The blood group A determinant is carried by four basic carrier carbohydrate chains. This paper reports the expression of blood group A variants, as determined by immunohistology with highly specific monoclonal anti-A antibodies, in 18 adenocarcinomas of the distal colon, 4 specimens of normal proximal mucosa from group A persons, and 5 specimens of fetal colonic mucosa. Monoclonal antibodies directed to type 1 chain A, type 1 chain ALeb, type 2 chain A, type 2 chain ALey, and type 3 chain A (repetitive A) were used. In normal mucosa, type 1 chain A and ALeb were expressed in proximal regions. Type 1 chain A was expressed in columnar cells, whereas type 1 chain ALeb was found in goblet cells. Type 2 and type 3 chain A structures were not found in normal adult mucosa. All types of A antigens were detected in adenocarcinomas from the distal colon as well as in normal fetal mucosa. In fetal mucosa, type 1 chain A and ALeb antigens and type 3 chain A antigens were expressed in columnar cells, whereas type 2 chain A and Ley and type 1 chain ALeb antigens were found in goblet cells. The results indicate that blood group A antigens with type 1, 2, and 3 carriers are present in fetal mucosa and adenocarcinomas of distal colon, while epithelial mucosa of normal adult colon is characterized by the exclusive expression of type 1 chain A antigens.     

Immunohistochemical detection of MAGE tumor-associated antigens in esophageal squamous cell carcinoma

Genes of the MAGE family encode tumor-specific antigens recognized by cytotoxic T-lymphocytes in a variety of neoplasms. We investigated the protein expression of these antigens as related to the gene expression, in esophageal squamous cell carcinoma by using monoclonal antibodies recognizing MAGE gene products. Esophageal squamous cell carcinomas were found to express both MAGE-1 (4 out of 15 samples) and MAGE-3 (7 out of 15 samples) genes, by RT-PCR. Immunoblotting revealed MAGE-1 and MAGE-3 gene products in 2 and 6 out of 15 samples, respectively. Immunohistochemistry performed on 12 samples showed MAGE-1 protein expression, limited to single tumor cells, in 2 cases. MAGE-3 gene product was detectable in 7 cases: in 5 of them over 50% of neoplastic cells were positive. Considering the high percentages of tumor cells expressing MAGE-3 antigen, the use of epitope-based vaccines could be envisaged in patients displaying appropriate HLA-class I phenotype.     

Blood group-related antigens in human kidney: modulation of Lewis determinants in renal cell carcinoma

Seven mouse monoclonal antibodies and the lectin Ulex europaeus, detecting blood group related antigens of the ABH and Lewis systems, have been used to determine the immunophenotype of human renal cell carcinomas. Immunohistochemical analyses have demonstrated that these antigenic systems are differentially expressed by distinct normal cell types and domains of the human nephron. In the present study we analyzed the immunophenotype of 29 primary and 15 metastatic renal carcinomas by the immunoperoxidase method. Blood type was known in all of the cases and secretor status in nine cases. ABH specificities were not detected in tumor cells of the primary tumors studied, although two of the metastases showed heterogeneous expression of H and A antigens, respectively. Lewisx (Lex) determinant was detected in 76% of primary renal cell carcinomas; however, Lex was only expressed by occasional cells in 20% of the metastatic tumors analyzed. Lewisa (Lea) was detected with a heterogeneous pattern of expression in 31% of the primary and 26% of the metastatic renal tumors studied. Lewisy (Ley) antigen expression was found in 17% of the primary and 20% of the metastatic tumors analyzed. Detection of precursor type 1 structure was observed in 28% of primary and 20% of metastatic renal cell carcinomas. The present study suggests the histogenesis of renal cell carcinoma in the proximal nephron, based on the expression of Lex and Lea antigens. It also shows: (a) an apparent deletion, downregulation or structural modification of Lex determinant in most of the metastatic tumors; (b) undetectable levels of ABH specificities in tumor cells of primary renal cell carcinoma; and (c) enhanced expression and/or neosynthesis of precursor type 1 structure and Ley determinant in some renal cell carcinomas.     

Full expression of blood group-related, transplantation-related, and carcinoembryonic antigens in human colorectal cancer cells with different degrees of phenotypic differentiation

Seven established human colon carcinoma cell lines with distinct degrees of phenotypic differentiation were evaluated for the presence of blood group-related and transplantation-related antigens in relation to their production of carcinoembryonic antigen (CEA). All lines presented A and B antigens regardless of the patients' original red blood cell type. However, tumor cells from patients originally classified as O-type had lower expression of both A and B antigens and high production of CEA. Cells from patients with an original A type had low to undetectable CEA production and high expression of both A and B antigens. There was no particular segregation of transplantation-related antigens with respect to phenotypic expression. All lines presented HLA-A, -B, and -C, as well as -DR antigens. These results demonstrate that colon carcinoma cells have the ability to fully express both blood group-related and transplantation-related antigens, even if discordant with the donor's red blood cell phenotype. Furthermore, it appears that expression of A antigen is intimately related to synthesis of CEA.     

Immunohistochemical demonstration of tumor-associated antigens in urinary bladder carcinomas using mono- and polyclonal antisera

Keratin was found in more than 90% of transitional cell carcinomas of the bladder in the cytoplasma with polyclonal antibodies. Intensity increased with dedifferentiation. Cytokeratin was detected with monoclonal antibodies in more than 80%. Squamous cell carcinoma of the urinary bladder was always strongly positive for keratin and cytokeratin. CEA was found in 20% of G1 and 40% of G2 and G3 carcinomas of the urinary bladder. The prostatic epithelium markers PSA and PAP were always negative also Ca1.     

Establishment of hamster pancreatic ductal carcinoma cell line (PC-1) producing blood group-related antigens

A pancreatic cancer cell line, PC-1, was established from a pancreatic ductal carcinoma induced in a hamster by N-nitrosobis(2-oxopropyl)amine (BOP). The cells grew in a monolayer with a doubling time of 38 h, and floated or piled up to form a duct-like structure. Chromosome counts ranged from 42 to 89. Light and electron microscopic studies of PC-1 cells revealed production of conspicuous amounts of amorphous substance. Injection of PC-1 cells into the homologous hamster pancreas resulted in tumor formation, histopathologically indistinguishable from the original primary pancreatic ductal carcinoma. Immunohistochemical expression of blood group-related antigens (BGRAs), A, B, H, Leb, Lex and Ley was observed both in the cells in the culture, and in tumor transplanted into the pancreas. In the culture supernatant, a high titer of blood group A antigen was detected. This cell line may provide a unique tool for studying the mechanism of BGRA synthesis and release in malignant cells.     

Immunohistochemical detection of carcinoembryonic antigen in esophageal carcinomas: a comparison with other gastrointestinal neoplasms

BACKGROUND: Although the prognostic value of Carcinoembryonic antigen (CEA) in colorectal cancer follow-up is well known, CEA expression in esophageal cancer is not widely recognized and studies correlating tissue CEA expression in stomach cancers with tumor differentiation have yielded contradictory results. We compared the CEA expression in esophageal, gastric and colorectal carcinomas in order to elucidate its diagnostic and prognostic significance and to evaluate the potential role of tissue CEA localization for the clinical evaluation of esophageal carcinomas. MATERIALS AND METHODS: CEA expression in 84 biopsies of carcinomas of the gastrointestinal tract (38 colorectal, 22 gastric, 24 esophageal) was evaluated by immunohistochemistry. RESULTS: Sixty-two percent of the squamous carcinomas of esophagus, all five esophageal adenocarcinomas arising in Barrett esophagus, 86 percent of gastric adenocarcinomas and 89 percent of colorectal adenocarcinomas were CEA-positive. In colorectal and gastric adenocarcinomas, CEA staining was present, usually at the luminal membrane of neoplastic glands and in intraluminal material. Signet ring cells were strongly positive for CEA. In esophageal carcinomas staining was mostly cytoplasmic and in the better differentiated tumors it was particularly prominent at the center of epithelial pearls. Intensity of staining was highest in well-differentiated carcinomas and lowest in poorly-differentiated carcinomas. CONCLUSION: A clear correlation was seen between the degree of tumor differentiation and CEA expression for carcinomas of the esophagus, stomach and colon. Our results support the potential usefulness of CEA for monitoring the recurrence of gastric or esophageal tumors. Immunohistochemical determination of tumor CEA content could be a useful adjunct for the management of gastrointestinal carcinomas, by improving interpretation of serum CEA levels.     

Amplification of the hst-1 gene in human esophageal carcinomas

The hst-1 gene, previously designated as the hst gene, and seven other oncogenes were examined for possible structural changes in esophageal, gastric and colorectal carcinomas by Southern blot hybridization. The hst-1 gene was amplified in eight (42.1%) of the nineteen esophageal squamous cell carcinomas and in all four metastatic tumors of lymph nodes. The degree of amplification ranged from two to eight times. Coamplification of the hst-1 and c-erbB-1 gene was found in one case of esophageal carcinoma. However, no amplification of the hst-1 gene was detected in gastric and colorectal carcinomas.     

Immunohistochemical studies on Waf1p21, p16, pRb and p53 in human esophageal carcinomas and neighboring epithelia from a high-risk area in northern China

To better understand the roles of p53 and cell cycle–regulating protein alterations in human esophageal carcinogenesis, we investigated immunohistochemically the distribution patterns of Waf1p21, pRb, p16 and p53 in 22 cases of surgically resected esophageal cancer as well as in the neighboring non-cancerous squamous epithelia. Waf1p21 protein was detected in 13 of the 20 cases of well-differentiated squamous-cell carcinoma (SCC), where the Waf1p21-positive cells were located mainly in the interior layers of the cancer nests. Conversely, p53-positive cells were found mostly in the peripheral layers. Cells containing both Waf1p21- and p53-positive immunostaining were not observed in a double-immunostaining experiment. p16 was detected in both the nucleus and cytoplasm in 3 of the 22 cases of SCC. All of these p16-positive cancers showed an absence of pRb immunostaining; this result is consistent with the idea that expression of p16 is regulated negatively by pRb. Eleven of the 22 esophageal SCCs (50%) showed extensive pRb immunostaining cells, and the remaining 11 cases displayed a few pRb-positive cells or an absence of pRb immunostaining. In a majority of the morphologically normal squamous-cell epithelia samples, immunostaining of Waf1p21 and pRb was found in most of the cells in the parabasal layers (proliferation compartment), where PCNA-positive cells also resided. In the pre-cancerous lesions, Waf1p21 and pRb were detected in cells surrounding the top of the lesioned region, p16-positive cells were scattered in the basal cell hyperplastic and dysplastic lesions and p53-positive cells existed in 2 distinct patterns: “scattered” and “focal”. Int. J. Cancer 72:746–751, 1997. © 1997 Wiley-Liss, Inc.     

Cytogenetic studies on human breast carcinomas

Cytogenetic studies were performed on cell material obtained from surgical specimens of 50 human breast carcinomas and from 61 cancerous effusions of 46 patients. Classical cytogenetic analyses of numerical chromosome changes and marker chromosomes revealed the non-random involvement of chromosomes #X and #22 as monosomics, of chromosomes #3, #7, and #19 as trisomics, and chromosome #1 (particularly p 13 to q 12) in marker formation. Karyotypic evolution was followed in vitro and in vivo and showed a highly individualistic pattern of stability and variability.In addition, a systematic screening for the presence of cytogenetic equivalents of gene amplification (double minutes DM, homogeneously staining regions HSR) was carried out. A high incidence of DM-positive cases was detected in primary tumors (48%) as well as in metastatic cells from effusions (40%), with the frequency of DM-containing metaphases ranging from 1 to 100% in the positive cases. This finding supports the assumption of the fundamental biological importance of gene amplification in human solid tumors.Furthermore, chromosome breakage and micronuclei were observed in breast carcinoma cells as an apparent consequence of therapy-independent mutability.     

Chromogranin A-positive tumor cells in human esophageal squamous cell carcinomas

Gastrointestinal cancers have frequently shown neuroendocrine (NE) differentiation, but whether NE differentiation occurs in esophageal squamous cell carcinoma (ESCC) remains unclear. In this study, tissue sections obtained from 43 patients with ESCC from a high-incidence area of Northern China were used for the assessing of NE differentiation by immunohistochemistry using antibody against chromogranin A (CGA). In addition, the malignant characteristics and proliferation capacity of CGA-positive cells were also examined by immunohistochemistry. The clinicopathological significance of these CGA-positive tumor cells in ESCC was assessed. Of 43 ESCC samples, CGAimmunoreactive tumor cells were detected in 10 cases (23.26%). However, the CGA-positive tumor cells were scattered at a very low number among non-immunoreactive tumor cells and were rarely constituted a major part of cancer cell nests. Only 4.65% (2/43) cases showed a high density (>10 cells but <1% of total tumor cell mass) of CGA-positive tumor cells. P53 immunoreactivity was frequently shown, while Ki67 was hard to detect in these CGApositive cells. In addition, no relationship between CGA positivity rate and clinicopathological parameters was found. Thus, we concluded that lowdensity CGA-positive tumor cells can be detected in ESCC, supporting the notion that heterogeneous NE differentiation also exists in tumors that lack neuroendocrine cells in their normal epithelial counterparts.