Significance of alkaline phosphatase isozymes in the monitoring of patients with colorectal carcinoma

The clinical course of colorectal carcinoma may be monitored by tumor markers such as carcinoembryonic antigen (CEA), carcinoma antigen (CA) 19-9 and CA-50. Alkaline phosphatase isozymes were previously used to study the clinical course of testicular and gynecologic tumors. In this study we investigated 8 patients with advanced colorectal carcinoma. Their sera were analyzed for the tumor markers CEA, CA 19-9, CA-50 and three alkaline phosphatase isozymes: the nonspecific liver isozyme LAP, the intestinal isozyme IAP and the placental isozyme PLAP. Rising levels of CEA, CA 19-9 and CA-50 were seen as expected, and PLAP also showed rising levels during tumor progression. LAP remained elevated. This indicates an association between progression of colorectal carcinoma and a raised serum content of alkaline phosphatase isozymes.     

Levels of alkaline phosphatase isozymes in human seminoma tissue

The three human isozymes of alkaline phosphatases were quantitatively determined in normal testis and seminoma tissues. The highly selective assays were based on isozyme specific monoclonal antibodies. In the normal testis approximately 90% of the catalytic activity originates from the tissue unspecific alkaline phosphatase, and the remaining activity was due to trace expression of both intestinal (approximately 5%) and placental alkaline phosphatase (PLAP) or PLAP-like isozyme (approximately 5%). In homogenates of seminoma tissues, highly increased levels of all three isozymes were identified. Both the tissue unspecific alkaline phosphatase and PLAP-like enzymes displayed relative increases of 10- to 100-fold and intestinal alkaline phosphatase 2- to 10-fold compared with normal testis. This finding indicates that the entire genome coding for alkaline phosphatases may be activated in seminomas. The PLAP-like enzyme from seminoma cells comprises a heterogenous population of molecules demonstrating partial heat sensitivity and microheterogeneity upon starch gel electrophoresis in contrast to the pregnancy related PLAP. These findings have implications for the different PLAP assays used in the clinical monitoring of seminoma patients.     

Tumor Markers in Colorectal Carcinoma An Evaluation of Carcinoembryonic Antigen, Tissue Polypeptide Antigen, Placental Alkaline Phosphatase and Pseudouridine

The biologic markers carcinoembryonic antigen (CEA), tissue polypeptide antigen (TPA), placental alkaline phosphatase (PLAP) and pseudouridine were analysed in 37 patients with colorectal carcinoma. CEA, TPA and PLAP were derived from the serum and pseudouridine from the urine. The incidence of all four markers increased with advancing stages of the disease. Patients with distant metastases had elevated levels of CEA, TPA, PLAP and pseudouridine in 85, 27, 18 and 33 per cent of the total cases, respectively. When survival was compared, patients with 2 to 4 elevated markers had shorter survival than those with none or only one elevated marker.     

Histologic demonstration of antigens reactive with anti-p21 ras monoclonal antibody (RAP-5) in human stomach cancers

The immunohistochemical reactivity of RAP-5, a monoclonal antibody (MoAb) raised against a synthetic peptide corresponding to positions 10-17 of the ras gene product from T24 bladder carcinoma, was studied in 96 surgically resected stomach cancers of humans. The cytoplasm of cancer cells in 65 cases (68%) was positively stained with MoAb RAP-5, although the staining was heterogeneous among cancer cells. There was no definite correlation between depth of tumor invasion and reactivity to MoAb RAP-5. Cancer cells of poorly differentiated tumors showed a tendency to react less frequently and less intensely to MoAb RAP-5. In nontumorous gastric mucosa, parietal cells and some portions of intestinal metaplasia were stained with MoAb RAP-5. These findings suggest an increased expression of the ras gene product (p21) in about two-thirds of gastric adenocarcinomas and in some nonneoplastic gastric epithelial cells.     

Evidence for homology of normal and neoplastic human placental alkaline phosphatases as determined by monoclonal antibodies to the cancer-associated enzyme

The HeLa TCRC-1 human adenocarcinoma cell line expresses a form of alkaline phosphatase that is similar to the common S-variant of placental alkaline phosphatase (PLAP) on the basis of electrophoretic mobility, catalytic properties, and reactivity with polyclonal antibodies. More sensitive probes of changes in protein structure than polyclonal antibodies are monoclonal antibodies (MAbs) which recognize individual antigenic sites on molecules. Therefore, we produced MAbs to HeLa TCRC-1 cells and selected those which bound to the alkaline phosphatase expressed by the cancer cells. Seven MAbs were obtained and characterized by (a) fine specificity analysis using allelic variants of PLAP and other human alkaline phosphatase isozymes, (b) immunoglobulin isotype, and (c) relative binding affinities to PLAP from two sources, placental tissue and HeLa TCRC-1 cells. The seven MAbs bind the enzymes from both sources with equal affinity indicating a high degree of structural homology if not identity between the normal S-variant of PLAP and its cancer-associated counterpart. We note that most of the MAbs to cancer cell surface-bound PLAP express either Ig (immunoglobulin) G2a or IgG2b heavy-chain isotypes, a higher incidence of these classes of IgG than has been observed with the purified and soluble PLAP immunogen which yields MAbs predominantly of the IgG1 isotype. Finally, some of these antibodies, like the ones prepared from purified PLAP, recognize differences between allelic variants.     

Gonadotropin and alkaline phosphatase-producing gastric carcinoma with widespread metastasis to the bones

A case of advanced gastric carcinoma in a 47-year-old man with widespread metastasis of generalized bone and associated with high serum levels of human chorionic gonadotropin (hCG) and alkaline phosphatase (ALP) is reported. Macroscopically, the primary tumor was Borrmann type IV. Microscopically, it showed poorly differentiated adenocarcinoma. Bone metastasis, however, showed well-differentiated adenocarcinoma. These lesions did not manifest conspicuous trophoblastic differentiation. Immunohistochemical studies revealed that hCG-positive cells were observed sporadically in the primary tumor and lymph node metastasis, but remarkably in the bone metastasis. Cancer cells in the lymph nodes were positively stained by modified Burstone's ALP staining. The serum levels of hCG and ALP fluctuated with the clinical course. Therefore, the authors concluded that this gastric carcinoma produced hCG and ALP.     

Tumour and cellular localization by use of monoclonal and polyclonal antibodies to placental alkaline phosphatase

Monoclonal and polyclonal antibodies against placental alkaline phosphatase (PLAP) were evaluated for tumour immunolocalization of human PLAP-producing Hep 2 tumours in nude mice. The antibodies were labelled with 125I and injected i.p. in mice with developing Hep 2 tumours. The distribution of 125I-anti PLAP in various tissues showed that the labelled antibody was enriched in the tumour, the mean concentration ratio being 7.1 and 6.8 for polyclonal and monoclonal antibodies, respectively. A PLAP negative tumour (RD) showed a mean ratio of 1.2. There was a positive correlation between PLAP content and uptake of labelled antibody in the tumours. Hep 2 tumour cells in tissue culture showed 100% positivity for PLAP, while imprints of the tumour after passage in nude mice showed 40-50% positivity. PLAP offers potential as a useful marker for localizing tumours in humans.     

Patterns of seminoma tissue markers and deletions

Seminomas and control tissues were analyzed for several tumor markers. Very high levels of placental alkaline phosphatase (PLAP)-like enzyme levels were found in all 18 seminomas studied. The majority of the seminomas were of phenotype I, thus differing from palcental PLAP. The mean amount of enzyme protein as measured by monoclonal antibodies, was 100 times higher than in non-malignant tissues and 10 times lower than in placental tissue. The specific enzymatic activity in seminomas was about half of that observed in placenta. Similarly, the specific activity of PLAP-like enzymes in sera of patients with seminoma was only about half of that found in pregnancy sera. HCG was strongly elevated in 3 seminomas, but not obviously related to PLAP. Thirteen of the 17 pure seminomas had HCG over 100 IU/g, which was not seen in normal testes. Liver alkaline phosphatase (LAP) and intestinal alkaline phosphatase (IAP) were high in seminomatous tissues, the mean increases being 60-fold and 20-fold, respectively. The highest IAP levels were found in 2 yolk-sac tumors. Ferritin was moderately elevated in seminomas, but high in several control tissues. Carcinoembryonic antigen (CEA) was not elevated and alpha-fetoprotein (AFP) was not detected at all in pure seminomas. A decrease in carbohydrate antigen 50 (CA-50) content was noted in seminomas as compared to normal testes, yolk-sac tumors and choriocarcinomas. Defects in tumor-related enzymes may account for increase of PLAP and decrease of CA-50.     

人胎盘GST—π抗体的免疫组织化学检测对胃癌和胃癌前期病变的早期诊断意义

Human placental form of glutathione S-transferase (GST-pi) was detected in 38 human gastric carcinoma and 28 precancerous lesions by Avidin biotin peroxidase complex (ABC) method using anti-GST-pi antibody. Of 22 normal gastric mucosa, 91% were negative for GST-pi stain, 95% of 21 well and moderately differentiated gastric adenocarcinomas, all of 5 poorly differentiated gastric adenocarcinomas, 75% of undifferentiated gastric carcinomas, 85% of intestinal metaplasia and 100% of atypical hyperplasia were positive for GST-pi stain. These results indicate that GST-pi is a new sensitive marker for immunohistochemical detection of gastric carcinoma and precancerous lesion.     

Expression of Sulfated Carbohydrate Chains Detected by Monoclonal Antibody 91.9H in Human Gastric Cancer Tissues

Expression of sulfated carbohydrate chains in intestinal metaplasia (IM) and gastric cancer tissues was immunohistochemically evaluated by using a monoclonal antibody (MAb) 91.9H. While normal gastric tissues were negative for MAb 91.9H, IM and cancer tissues were positively stained with high frequency. The incidence was 67% for IM with chronic gastritis (n = 12), 95% for IM with gastric cancer (n=21), 77% for well and moderately differentiated adenocarcinomas (n = 13), 48% for poorly differentiated adenocarcinoma (n = 23), 89% for signet-ring cell carcinoma (n = 9) and 100% for mucinous adenocarcinoma (n = 7). When poorly differentiated adenocarcinoma cases were divided into two groups, solid (n = 13) and non-solid types (n = 10), the incidences were 8% and 100%, respectively. These data suggest that MAb 91.9H could be of practical use as a new marker for IM and gastric cancer, and may be valuable for subgrouping poorly differentiated adenocarcinomas. Analyses of the core proteins for 91.9H epitope were then carried out. Comparison of immunostaining of ten poorly differentiated adenocarcinoma cases by MAb MUSE11 against MUC1 gene product with that by MAb 91.9H suggested that 91.9H epitope is not expressed on MUC1. Northern blot analysis of 10 pairs of gastric cancer and adjacent normal tissues with a MUC2 cDNA probe showed that the expression level of MUC2 mRNA was below the limit of detection. Thus, 91.9H epitope may be expressed on other proteins than MUC1 or MUC2 core proteins in gastric cancer.     

Detection of bone-type alkaline phosphatase by monoclonal antibodies reacting with human osteosarcoma-associated antigen.

The antigen detected by monoclonal antibodies reacting with human osteosarcoma-associated antigen was shown to be a phosphatidyl-inositol (PI)-glycan-anchored protein, which can be released from the cell surface by PI-specific phospholipase C-treatment. The antigen detected by 2D3 and 2H10 antibodies exhibited alkaline phosphatase activity. Both antibodies strongly reacted with bone-type alkaline phosphatase. However, importantly, immunohistochemical analysis demonstrated that 2D3 and 2H10 did not react with alkaline phosphatase present in kidney or liver. In addition, neither placental nor intestinal alkaline phosphatase was recognized by 2D3 and 2H10 antibodies. These results indicated that two monoclonal antibodies, 2D3 and 2H10, are highly specific for bone-type alkaline phosphatase and can distinguish bone alkaline phosphatase from liver alkaline phosphatase in spite of the fact that liver and bone alkaline phosphatase are encoded by the same gene.     

Detection of Bone-type Alkaline Phosphatase by Monoclonal Antibodies Reacting with Human Osteosarcoma-associated Antigen

The antigen detected by monoclonal antibodies reacting with human osteosarcoma-associated antigen was shown to he a phosphatidyl-inositol (Pl)-glycan-anchored protein, which can be released from the cell surface by PI-specific phospholipase C-treatment. The antigen detected by 2D3 and 2H10 antibodies exhibited alkaline phosphatase activity. Both antibodies strongly reacted with bone-type alkaline phosphatase. However, importantly, immunohistochemical analysis demonstrated that 2D3 and 2H10 did not react with alkaline phosphatase present in kidney or liver. In addition, neither placental nor intestinal alkaline phosphatase was recognized by 2D3 and 2H10 antibodies. These results indicated that two monoclonal antibodies, 2D3 and 2H10, are highly specific for bone-type alkaline phosphatase and can distinguish bone alkaline phosphatase from liver alkaline phosphatase in spite of the fact that liver and bone alkaline phosphatase are encoded by the same gene.     

Special features of intestinal metaplasia and its relation to early gastric carcinoma in man: observation by a method in which leucine aminopeptidase activity is used.

The degree of intestinal metaplasia and its macroscopic distribution in gastric mucosa were examined by a new method in which leucine aminopeptidase (LAP) activity is used to investigate the relationship between intestinal metaplasia and gastric carcinoma. Since LAP is a specific enzyme of intestinal metaplasia, the area showing positive reaction for this enzyme corresponds strictly with the zone of intestinal metaplasia examined microscopically. Almost all metaplasia was demonstrated by this method. This method was used to examine 40 human gastric carcinomas confined to the mucosa. Gastric mucosa containing differentiated tubular-type carcinoma (60%) was associated with a high degree of metaplasia in comparison with the low degree of metaplasia in poorly differentiated carcinoma (40%). Differentiated tubular-type carcinoma was closely related to intestinal metaplasia. However, all carcinomas arising from the mucosa without intestinal metaplasia (18%) were poorly differentiated. Therefore, gastric carcinomas occurring in Japanese patients are frequently and closely related to intestinal metaplasias.     

Production and characterization of a monoclonal antibody (MSN-3) for uterine endometrial and endocervical adenocarcinoma

A monoclonal antibody (MSN-3) was raised using HEC-108 cells derived from poorly differentiated endometrial carcinoma as the immunogen. The immunoglobulin subclass of MSN-3 was IgGr1. The target antigen of MSN-3 was a protein with a molecular weight of 77 kDa, and it was shown to be localized in the cytoplasm. MSN-3 only reacted with 14% of normal proliferative endometrium cells, but it showed a high positivity rate of 66% for endometrial carcinoma. The target antigen of MSN-3 increased as endometrial cells became more malignant, and the possibility of changes in localization was also suggested. Moderately and poorly differentiated endometrial carcinoma showed a high positivity rate for MSN-3. MSN-3 reacted rarely or not at all with normal cervical glandular tissue, but the positivity rate for cervical adenocarcinoma (especially endocervical adenocarcinoma) was a high rate of 59%. The patterns of staining of endocervical adenocarcinoma by MSN-3 included diffuse staining of the whole cytoplasm and not only that near the glandular lumen, as well as staining of the basal cytoplasm. Changes in the localization of the target antigen were clearly associated with carcinogenesis of the cervical glandular cells. The MSN-3-positive rate was high in patients with lymph node metastasis and vascular invasion. Among the staining patterns, the basal and diffuse patterns tended to increase with malignacy. The basal pattern of staining was characteristic of MSN-3, suggesting that it might assist in the diagnosis of cervical adenocarcinoma.     

Electrophoretic heterogeneity of alkaline phosphatase isozymes in seminoma and normal testis

Electrophoretic patterns of seminoma- and normal-testis-derived alkaline phosphatase isozymes, the placental alkaline phosphatase (PLAP)-like enzyme and the tissue-nonspecific (liver) alkaline phosphatase (LAP), were studied on starch gel and isoelectric focusing (IEF). Different migration patterns of the PLAP-like enzyme were observed with respect to both seminomas and normal testes on starch gel electrophoresis. On IEF, seminomas showed different staining patterns among different tumors; however, a common main activity was focused at pIs of 4.3-4.6, corresponding to pIs of PLAP. Normal testes showed two enzyme-staining regions, at pIs of 4.1 and 5.0-5.2, which were discriminated from pIs of PLAP and the PLAP-like enzyme in seminoma. The PLAP-like enzyme in seminoma was differentiated from PLAP by digestion with neuraminidase. Neuraminidase treatment simplified the distribution patterns of the PLAP-like enzyme in normal testis, but did not alter the pattern of microheterogeneity in seminoma. Two factors other than sialylation, namely structural modification of the carbohydrate moiety and variation of hydrophobicity, were shown to contribute to the microheterogeneity of the PLAP-like enzyme in seminoma. LAP in seminoma and in normal testis also showed marked electrophoretic heterogeneity and differences in pI distributions from LAP of liver. However, the migration patterns after desialylation were very similar to each other. The findings imply that electrophoretic heterogeneity demonstrated in LAP in seminoma and in normal testis is caused by a difference in sialic acid content in the molecule, and the heterogeneity of the PLAP-like enzyme in seminoma is considerable.     

Expression of p-glycoprotein in the normal colorectal epithelium and in colorectal-carcinoma

Expression of P-glycoprotein (pgp) was analyzed by immunohistochemical staining with monoclonal antibody JSB-1 in 145 frozen specimens (67 were samples of normal colorectal mucosa from sites adjacent to the tumor, 66 were colorectal carcinomas, 5 colorectal polyps, 5 metastatic lymph nodes, and 2 samples of metastatic liver tumors) of 67 patients with colorectal carcinoma and polyps. All 72 specimens of normal colorectal mucosa and adenomatous polyps expressed pgp to various degrees. By contrast, 18 of 39 (46.2%) samples from cases of well differentiated adenocarcinoma were positive for pgp but only 3 of 21 (14.3%) samples from cases of moderately differentiated adenocarcinoma and none of the 4 samples from cases of poorly differentiated adenocarcinoma were positive for pgp. There was no correlation between the clinicopathological stage of colorectal carcinoma and the expression of pgp. These findings indicate that the expression of P-glycoprotein is closely related to the differentiation of cells. In normal colorectal epithelium, pgp was expressed normally and in well differentiated adenocarcinomas, pgp was still expressed. However, expression of pgp was no longer detectable in carcinomas with moderate or poor differentiation.     

Differential expression of peroxisome proliferator-activated receptor in histologically different human gastric cancer tissues

Gastric cancer cell lines express peroxisome proliferator-activated receptor gamma (PPARgamma), and treatment with PPARgamma ligands suppresses growth of subgroup of these cell lines. However, expression and subcellular distribution of PPARgamma in human gastric cancer tissues is still unknown. Therefore, expression and subcellular localization of PPARgamma were examined among different histological types of gastric cancer tissues. Immunohistochemical staining for PPARgamma was performed using biopsy specimens of human gastric cancer of various histological types, gastric adenomas, and intestinal metaplasia. All samples of intestinal metaplasia and most samples of gastric tumors, except for signet ring cell carcinoma, expressed PPARgamma in the epithelial cells. Most samples of signet ring cell cancer lacked PPARgamma expression. All samples of intestinal metaplasia expressed PPARgamma only in the cytosol. For adenoma, 90% was positive for PPARgamma in cytosol, and 40% was positive in nuclei, for well-differentiated adenocarcinoma, 80% was positive in cytosol, and 20% was positive in nuclei. For moderately differentiated adenocarcinomas, 70% was positive for cytosol, and 80% was positive for nuclei; for poorly differentiated adenocarcinoma, 30% was positive in cytosol, and 70% was positive in nuclei. The frequency of samples with positive cytosolic staining decreased as the differentiation stage turned from intestinal metaplasia to adenoma, well-, moderately-, and poorly-differentiated cancers. Simultaneously, there was a tendency toward an increased frequency of samples with positive nuclear PPARgamma staining as the differentiation stage transformed from intestinal metaplasia to poorly-differentiated cancer. There was a striking difference in subcellular localization according to the differentiation levels of gastric dysplastic cells. The findings also supported an intestinal metaplasia-adenoma-well-differentiated gastric cancer sequence, and signet ring cell cancer was suggested to be of a different lineage from other types of gastric cancers.     

不同分化程度胃癌的多排螺旋CT表现特点

目的 探讨不同分化程度胃癌的多排螺旋CT表现特点.方法 收集经手术病理证实的230例胃癌患者,分析不同分化程度胃癌的病灶大体形态、病灶强化程度及淋巴结CT影像学特点,并与病理结果 对照.结果蕈样型胃癌181例(78.70%),溃疡型胃癌24例(10.42%),息肉型胃癌18例(7.83%),弥漫型胃癌7例(3.05%).胃癌诊断准确率为98.26%.弥漫型胃癌中没有高分化腺癌,低分化腺癌和中分化腺癌共占82.35%.CT大体形态不同分化程度胃癌差异无统计学意义(P>0.05).胃癌病灶平扫及动脉期CT值各分化组间差异无统计学意义(P>0.05),静脉期及延迟期各分化组间差异具有统计学差异(P<0.05).高分化胃癌病灶的CT值在静脉期和延迟期明显小于低分化和中低分化腺癌(P<0.05).通过测量淋巴结短径CT共检出≥6 mm淋巴结556枚,≥8 mm淋巴结290枚,≥10 mm淋巴结229枚.不同分化程度胃癌组织淋巴结平扫及三期增强CT值比较,差异无统计学意义(P>0.05).结论 CT中低分化及低分化胃腺癌病灶的强化程度,高于高分化及中分化胃腺癌,强化峰值在静脉期和延迟期.     

ING2和mP53蛋白在胃癌组织中的表达及临床病理意义

目的 检测生长抑制因子2(ING2)蛋白在正常胃组织、胃癌及其癌前病变组织中的表达,分析ING2与突变型P53(mP53)在胃癌中表达的关系,并探讨ING2与胃癌发生发展的关系及临床病理学意义.方法 免疫组织化学PV9000二步法检测188例胃癌及128例配对正常胃黏膜、35例慢性萎缩性胃炎、87例肠上皮化生及36例异型增生组织中ING2的表达.取其中的40例胃癌组织同时检测mP53蛋白的表达.结果 ING2蛋白在慢性萎缩性胃炎(74.29%)、肠上皮化生(91.95%)、异型增生(75.00%)和胃癌组织(70.21%)中阳性率均显著高于正常胃黏膜(36.72%),P＜0.001.ING2蛋白在胃癌中的表达与Lauren分型有关,在肠型胃癌中的表达率(80.56%)显著高于弥漫型(64.49%)和混合型胃癌(55.56%),P＜0.05,而且ING2蛋白在高-中分化管状腺癌的表达率(80.60%)显著高于低分化管状腺癌的阳性表达率(62.62%),P＜0.05.胃癌组织中ING2与mP53蛋白表达无相关性,P＞0.05.结论 ING2蛋白的过表达及细胞定位的改变可能参与胃癌的发生和分化,尤其是肠型胃癌的发生.     

Prognostic significance of p53 overexpression in gastric and colorectal carcinoma.

p53 expression was examined in 55 gastric and 107 colorectal carcinomas with an immunoperoxidase technique, using the polyclonal antibody CM1 on routinely fixed, paraffin embedded tissue. p53 protein was detected in 47% gastric and in 46% colorectal carcinomas and found to correlate with stage of disease and unfavourable clinical outcome (P less than 0.001). Thus, the proportion of positively reacting neoplasms increased as the stage progressed, tumours which had invaded regional lymph-nodes overexpressed p53 more frequently than localised carcinomas and an elevated level of p53 was associated with early relapse and death. In colorectal carcinoma p53 positivity was also linked with site and macroscopic configuration of the primary tumour and was most frequently expressed in carcinomas from the rectum and in ulcerative tumours. p53 overexpression was irrespective of tumour grade. Uniform negative reactivity with anti-p53 antibody was seen in normal epithelium adjacent to carcinoma, intestinal metaplasia, atrophic gastritis and in colonic adenomas. There was a good correlation between immunohistochemical staining on paraffin and frozen sections. These studies suggest that in gastric and colorectal carcinoma, immunohistochemical detection of p53 protein in routinely fixed tissue can be used along with other established parameters to assess prognostic outcome, especially to identify patients with poor short-term prognosis.