Independent prognostic value of ploidy in colorectal cancer. A prospective study using image cytometry.

In a prospective study, the DNA content of Feulgen-stained nuclei obtained from fresh samples of 211 colorectal adenocarcinomas was evaluated by means of image analysis. The DNA histogram classification took into account aneuploidy and S-phase fraction for diploid cases. No significant relationship was found between ploidy and sex, age, preoperative carcinoembryonic antigen (CEA), size of the tumor, histologic differentiation, or Dukes' stage. Aneuploidy was more frequently encountered in distal tumors. Preoperative CEA, histologic differentiation, Dukes' stage, and ploidy were individually associated with overall survival. In Dukes' A, B, and C tumors, patients with normal and elevated CEA had no significant difference in overall survival. A relationship was apparent between disease-free survival and site, histologic differentiation, Dukes' stage, and ploidy. Multivariate overall survival analysis did not reveal independent prognostic significance of ploidy when all Dukes' stages were considered. In contrast, Dukes' stage, differentiation, and ploidy were good indicators of higher risk of colorectal cancer-related death in patients undergoing curative surgery. Dukes' stage and ploidy were also indicators for recurrence. Thus, routine histopathologic characteristics should be used in combination with quantitative cytologic features for the definition of a relevant prognostic index in colorectal cancer.

Characterization of the shed form of the human tumor-associated glycoprotein (TAG-72) from serous effusions of patients with different types of carcinomas.

Monoclonal antibody (MAb) B72.3 binds a high molecular weight tumor-associated glycoprotein designated TAG-72. This study reports the isolation and characterization of secreted TAG-72 directly from effusions of ovarian, colorectal, pancreatic, and endometrial carcinoma patients and compares them to TAG-72 derived from the LS-174T colon carcinoma xenograft. The B72.3-reactive antigen, TAG-72, was used as immunogen to produce second generation anti-TAG-72 MAbs. One of these second generation MAbs, CC49, had a higher affinity than that of B72.3 and was utilized as an affinity reagent in a procedure to purify the TAG-72 present in the serous effusions of carcinoma patients. A three-step purification procedure, utilizing heat extraction, CC49 antibody affinity chromatography, and gel filtration chromatography, resulted in 1000- to 4400-fold purifications of the TAG-72 derived from effusions, as analyzed using a double-determinant radioimmunoassay. Radiolabeled TAG-72 from each of the effusions demonstrated similar high molecular weight bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Similar results from the various effusions were also obtained in Western blotting analyses. Analyses of TAG-72 from the different effusions in radioimmunoassay using five different anti-TAG-72 MAbs revealed similar binding patterns. The results of these studies thus indicate that TAG-72 obtained directly from patients with ovarian, colorectal, endometrial, and pancreatic carcinomas and the LS-174T xenograft are highly similar in terms of immunochemical properties and antigenic profile.

[Immunocytochemical study of enterochromaffin cells in carcinoma of the large intestine].

In a series of 130 cases of adenocarcinoma of the large intestine, enterochromaffin (EC) cells were detected in 54 cases (41.5%) by immunocytochemistry with anti-chromogranin monoclonal antibody. Among the 54 cases, 30 were found positive for serotonin, 14 for somatostatin, 11 for glucagon, 5 for pancreatic polypeptide, and only one for gastrin. The cases with EC cells (++) or polypeptide positive cells exhibited higher grade of differentiation, earlier stage of tumour extension and higher survival rate than those without EC cells. A significant difference of the EC cell population pattern among different histological grades of the tumours and nonneoplastic mucosa was found. The proportion of hormone, especially polypeptide positive cells was the highest in the mucosa and lowest in the moderately poorly differentiated carcinomas. The incidence, methodology and clinicopathological significance of EC cells found in the tumours are discussed.

Serotonin and 5-hydroxyindoleacetic acid are increased in the sigmoid colon in severe idiopathic constipation.

The distribution of serotonin and dopamine beta-hydroxylase was examined in sigmoid colon specimens from patients with severe idiopathic constipation and control patients with carcinoma of the rectum or colon. Specimens were divided into three regions: (a) the mucosa; (b) the myenteric and submucosal plexuses with the longitudinal and circular smooth muscles; and (c) the circular smooth muscle, for biochemical analysis of serotonin and 5-hydroxyindoleacetic acid (total indoles) and noradrenaline. In both groups of patients, serotonin- and dopamine beta-hydroxylase-like immunoreactivity was localized in nerves in the myenteric and submucosal plexuses, and a sparse innervation was observed in the circular muscle. In addition, intense serotonin-like fluorescence was present in a large number of enterochromaffin cells in the mucosa. Total indole levels were significantly increased in the mucosa (p less than 0.02) and circular muscle (p less than 0.05) of the constipated patients. In contrast, no changes in noradrenaline levels were observed in any of the regions studied. Altered levels of total indoles may thus contribute to severe idiopathic constipation. Analysis of biopsy specimens could be a useful tool in clinical diagnosis and future investigations of diseases of the gut.

Rectal carcinoid tumor metastasizing to the thyroid and pancreas. An autopsy case exploiting immunohistochemistry for differentiation from tumors involving multiple endocrine organs.

A 58-year-old male patient with rectal carcinoid tumor is presented. The tumor extensively involved the lymph nodes and liver, and multiple tumors were also recognized in the pancreas and thyroid. Grossly, it was uncertain whether the latter were metastases from the rectal carcinoid or all were coincident primary tumors involving multiple endocrine organs, so-called multiple endocrine neoplasia (MEN) syndrome. Histologic, histochemical and electron microscopic examinations of the tumors in both the pancreas and thyroid showed similar features to those of the rectal carcinoid. The neoplastic cells in all involved organs commonly expressed positive immunoreactivity for somatostatin, but negativity for carcinoembryonic antigen, calcitonin, calcitonin gene-related peptide, thyroglobulin, insulin, glucagon and pancreatic polypeptide. These immunohistochemical results confirmed that the tumors observed in multiple endocrine organs were indeed metastatic from the rectal carcinoid, rather than being a new combination of MEN syndrome. Some neuroendocrine tumors may develop widespread metastasis, sometimes creating problems with differentiation from multiple primary endocrine tumors. Immunohistochemistry may be of great help in setting this issue.

Ultra-sonografia transretal no estadiamento do câncer de reto: estudo inicial / Intrarectal ultrasound in the staging of rectal cancer: a preliminary study

O prognóstico do carcinoma do reto está ligado à extensäo da lesäo na época do diagnóstico. Um adequado estadiamento pré-operatório do câncer do reto permitirá que o tratamento correto seja planejado. A ultra-sonografia transretal tem se mostrado eficaz em determinar o grau de infiltraçäo do tumor na parede do reto, a invasäo da gordura e dos linfonodos perirretais e a invasäo de órgäos vizinhos. Para comparar a capacidade da ultra-sonografia transretal em demonstrar a extensäo da neoplasia, 14 pacientes com carcinoma de reto provado foram examinados antes da cirurgia, e, os resultados, comparados com o exame das peças cirurgicas. O exame endossonográfico estadou corretamente 12 dos 14 tumores, ocorrendo umfalso negativo. No outro paciente, o exame näo pode ser concluido porque o transdutor näo conseguiu transpor a area do tumor. Seis outros pacientes em seguimento pós-cirurgico por cancer de reto foram submetidos a endossonografia, demonstrando-se recidiva local em três. Uma revisäo do padräo sonográfico da paredo do reto normal é feita, com atençäo especial para a correlaçäo entre as camadas histológicas e as sonográficas. Baseado nobaixo custo, na acuracidade e simplicidade, a ultra-sonografia transretal é proposta como um método que permite adequado estadiamento do cancer de reto.

Oestrogen receptors in colorectal carcinoma.

The oestrogen receptor content of colorectal adenocarcinoma was investigated using an established ligand binding biochemical assay and two more recently introduced techniques using specific monoclonal antibodies (Abbott ER-EIA and ER-ICA assay kits). Twenty nine tumours were investigated by the ligand binding assay. Only one (3.4%) tumour gave a weakly positive result (11 fmol/mg cytosol protein); the rest were all negative. Where sufficient tissue was available, the receptors were also determined by a quantitative immunoassay in 18 patients and an immunohistochemical method in 13 patients. The results were similarly all negative. It is concluded that most colorectal carcinomas, irrespective of sex, are oestrogen receptor negative, and it is thus unlikely that hormonal manipulation would have an influence on the course of the disease.

Significance of DNA content abnormalities in small rectal cancers.

Several studies have shown that the presence of DNA content abnormalities, measured by flow cytometry, may correlate with a poor prognosis in a variety of cancers. The predictive value of DNA content in patients with small rectal cancers has not been well determined. Thirty-nine patients with primary rectal adenocarcinoma smaller than 3 cm were studied in a comparison of DNA content with established prognostic variables. The following covariates were evaluated for their prognostic value: sex, age, tumor size, location, distal margin, Dukes' classification, tumor differentiation, and DNA content. DNA content was assessed by flow cytometric analysis, and each tumor was categorized as diploid or nondiploid. Of the parameters studied, Dukes' classification and tumor DNA content were found to be independent prognostic indicators. Determination of DNA content seems to provide additional useful prognostic information in patients with small rectal tumors.

Polyamines in colorectal cancer. Evaluation of polyamine concentrations in the colon tissue, serum, and urine of 50 patients with colorectal cancer.

Total, free, and acetylated polyamine concentrations were measured simultaneously in colon tissue, serum, and urine of 50 patients with histologically proven colorectal cancer, 40 patients with nonmalignant gastrointestinal diseases, and 30 healthy volunteers. Compared with histologically unaffected colon tissue, concentrations were significantly (P less than 0.001) higher for putrescine, elevated for cadaverine, and nearly identical for spermidine and spermine in colon carcinoma, whereas N1-acetylated and N8-acetylated spermidine were detectable in cancer tissue only. Serum and urine concentrations of all polyamines except total cadaverine and spermine in serum and free spermine in urine were significantly elevated compared with healthy controls and highest sensitivity for colon cancer was found for total spermidine (89.15%) in serum and acetylputrescine (84.5%), total putrescine (84.0%), N1-acetylspermidine (79.3%), and total spermidine (92.1%) in urine. However, nonmalignant gastrointestinal diseases partly showed similar elevations which resulted in a low specificity for polyamines in colorectal cancer. Therefore, polyamines are of little value only as diagnostic markers in colorectal carcinoma. Since polyamine concentrations in serum and urine normalized in patients after curative operation while they were further elevated in patients with proven tumor relapse or metastases, these substances might play a clinical role in predicting therapeutic success or indicating relapse of the tumor. Although a significant dependency of polyamine concentrations in serum or urine to Dukes' classification, tumor localization, CEA, CA 19-9, or CA 125 did not exist, a significant linear correlation was found for tumor size.

[The characteristics of bile acid excretion in patients with cancer of the large intestine]. / Osobennosti ékskretsii zhelchnykh kislot u bol'nykh rakom tolstoi kishki.

Marked difference in bile acids excretion (on the same diet) was observed between patients with large bowel cancer and healthy controls. The peak level of the bile acids in feces was registered in female patients 46-59 years of age suffering right-sided cancer. It is suggested that bile acid excretion in man is determined both by nutritional factors and hormonal-metabolic peculiarities of the body. High excretion of the bile acids is associated with risk for colon cancer.

Rectal and colonic carcinoids. A clinicopathologic study of 84 cases.

Eighty-four carcinoids of the colon and rectum were studied with emphasis on prognostic features, immunohistochemical characteristics, and pitfalls in diagnosis. Follow-up data were available on 35 patients. Tumors with adenocarcinomatous components, or those resembling small cell carcinomas of the lung, were excluded. Eighty-one tumors were in the rectum and three tumors were in the distal sigmoid colon. Neuron-specific enolase, chromogranin, and Leu-7 were positive in 87%, 58%, and 53% of the tumors, respectively. Hormones were positive in the following percentages: serotonin, 45%; pancreatic polypeptide, 46%; glucagon, 10%; gastrin, 3%; somatostatin, 3%; adrenocorticotrophic hormone, 1%; cholecystokinin, 0%; calcitonin, 0%; and insulin, 0%. Many tumors elaborated more than one hormone. Fifty-five percent of the tumors were argyrophil and 28% were argentaffin. Carcinoembryonic antigen was present in 24% of the tumors; 82% of the tumors contained prostatic acid phosphatase. Three patients had liver metastases; their tumors ulcerated, invaded muscularis propria, and had more than 2 mitoses per 10 high-power fields (HPF). One patient with a 2.5-cm tumor without mitoses had regional lymph node metastases. All non-metastasizing tumors had less than one mitosis in 10 HPF. We conclude that large bowel carcinoid tumors are essentially limited to the rectum and sigmoid, that they are indolent if mitotically inactive and smaller than 2 cm, and that most show production of a selected group of endocrine markers.

[Flow cytometric DNA analysis in rectal carcinomas].

The cellular DNA content was measured with flow cytometry from paraffin-embedded material in 329 patients and metastatic tumors of the liver from the rectum in 11 patients. The classification of the DNA ploidy pattern is as follows: A stem cell peak with a DNA index of 0.9-1.1 is defined as DNA diploid tumor and DNA aneuploid tumor is that with a DNA index greater than or equal to 1.1. There was a good correlation of DNA indices (r = 0.997) obtained from flesh and corresponding paraffin-embedded specimens. It is concluded that accurate determination of DNA index from paraffin-embedded materials is possible in the majority of cases. DNA ploidy of primary tumor cells correlated with clinicopathological findings such as lymphatic invasion, vascular invasion, lymph node metastasis and hepatic metastasis (p less than 0.01), but did not correlate with extramural carcinoma invasion. The cumulative survival rate (Kaplan-Meier) of curatively resected rectal carcinomas was worse in DNA aneuploid than in DNA diploid tumors (p less than 0.01). These observation showed that the determination of DNA ploidy in rectal carcinomas may prove to be of prognostic value.

Methane production in patients with colorectal carcinoma.

In searching for a screening test to identify a population at high risk for large bowel cancer, methane production was measured in 45 patients with colorectal carcinoma compared with 67 individuals who served as a control group. There was no significant difference in methane production between the colorectal cancer patients and the control group (37.8 and 25.4% respectively). Within the colorectal cancer group 54% of the males were methane producers compared with 19% of the females (P = 0.03). There were no differences according to disease stage. In view of these results, we see little value in using expired air methane concentration as a screening test for large bowel cancer.

Enzyme patterns and flow cytometric DNA measurements in colorectal hyperplastic polyps and tubular adenomas less than five millimeters.

Glucose-6-phosphate dehydrogenase (G6PD) activity, lactate dehydrogenase (LD) activity, and cytometric flow measurement of nuclear size and cell cycle distributions were registered in biopsy specimens from adenomas and hyperplastic polyps less than 5 mm. The G6PD and LD activities in adenomas, 36.8 +/- 4 U/g protein and 1580 +/- 163 U/g, respectively, were significantly higher than in hyperplastic polyps, 17.9 +/- 2.6 U/g and 1080 +/- 107 U/g, or in normal mucosa, 14.7 +/- 0.8 U/g and 1174 +/- 56 U/g, respectively. G6PD activity was significantly higher in adenomas with moderate (44 +/- 5.8 U/g) than in those with mild (32 +/- 5.9 U/g) dysplasia, in contrast to LD. No difference in enzyme activity was observed between hyperplastic polyps and normal mucosa. The growth of adenomas and hyperplastic polyps was not significantly different during the 2 years of follow-up study before biopsy specimens were taken. Increased nuclear size, unrelated to grade of dysplasia, was registered in 12 of 20 adenomas, in 1 of 10 hyperplastic polyps, and in 2 of 42 specimens from normal mucosa. Adenomas and normal mucosa from individuals with adenomas showed a significantly increased number of cells in S-phase. The values were 10.2 +/- 3% and 8.8 +/- 4%, respectively. Normal control value was 6.8 +/- 2.2%. An increased number of cells in G2-phase was observed only in adenomas (6.9 +/- 2.7% compared with 4.9 +/- 1% in controls). None of the adenomas showed aneuploidy.(ABSTRACT TRUNCATED AT 250 WORDS)

Specific receptors for 1,25-dihydroxyvitamin D3 (1,25-DR) and human colorectal carcinogenesis.

Because it is a common prerequisite for steroid responsiveness in target tissue, we investigated the presence of specific 1,25-DR in spontaneous human colorectal adenocarcinomas (ADC) and adjacent normal-appearing mucosa (NAM) from 23 operative specimens (12 male and 11 female patients). 1,25-DR was determined in cytosol by a DCC assay technique. 1,25-DR was present in 21 of 23 NAM and in only 4 of 23 HCRA. All positive ADC were well differentiated. Receptor content expressed in femtomoles/mg of protein (mean +/- SEM) was respectively 63.9 +/- 7.6 for right colon NAM and 51.3 +/- 12.9 for left colon or rectum NAM. When we compared all NAM specimens, receptor content was 56.7 +/- 8.0 femtomoles/mg of protein. No difference in 1,25-DR NAM level was observed between right colon and left colon or rectum. In adenocarcinoma the mean content was 66.5 +/- 14 fmoles/mg of protein. Scatchard analysis showed a single class of specific high-affinity saturable 1,25-DR with a dissociation constant (Kd) of 0.97 +/- 0.57 and 1.03 +/- 0.39 chi 10(-10) M in NAM and ADC respectively. These preliminary data represent the first demonstration of 1,25-DR throughout the entire human colon and indicate that the receptivity for this hormone is often lost during malignant transformation of the human colorectal mucosa. In addition, 1,25-DR could be a marker of differentiation in ADC. These preliminary results provide evidence supporting the addition of Vitamin D to the roster of developmental cancer chemopreventative agents.

Human chorionic gonadotropin in colorectal cancer and its relationship to prognosis.

The presence of human chorionic gonadotropin in large bowel cancers was studied immunohistochemically using an immunoperoxidase technique. HCG-positive tumour cells were present in 42 of 194 adenocarcinomas examined (22.0% of colon cancer and 21.2% of rectal cancers). On histological grading, the hCG-positive rate tended to rise as the degree of differentiation decreased. HCG was detected more frequently in cancers invading the total bowel wall (27%) than in those invading the partial wall (17.1%). Lymph node, liver or peritoneal metastases were present more frequently in hCG-positive tumours than in hCG-negative tumours. Furthermore, there was an intimate correlation between the presence of hCG-positive tumour cells and CEA doubling times in nine cases with untreated liver metastasis. The survival rate for patients with tissue hCG-positive cells was lower than for those with hCG-negative tumours. Thus, the presence of tissue hCG in colorectal cancers may be a biological marker of prognostic significance.

[Two cases of anorectal malignant melanoma].

Although on anorectal malignant melanoma is relatively rare, we report two cases, that of a 88-year-old woman, and a 78-year-old man. Both patients were admitted to the hospital due to rectal bleeding and were diagnosed as having rectal carcinoma. Following an abdominoperineal rectal amputation, a pathological examination of the resected specimens revealed a malignant melanoma. Thus, it is necessary to carry out a careful examination of patients with tumors of the anorectal region when there is the least suspicion of a malignancy.

Detection of ras gene alterations and ras proteins in colorectal cancer.

DNA extracted from 31 primary colorectal carcinomas was analyzed for the presence of ras gene amplification and mutations. Nine carcinomas had Ha-ras amplification and seven Ki-ras amplification. Nine carcinomas had codon 12 Ki-ras mutations. Immunohistochemical staining for ras proteins revealed a normal membrane association in normal mucosa and benign polyps but an abnormal cytoplasmic distribution in carcinomas. Amplification, mutations, and immunohistochemical staining were independent of histologic differentiation, Dukes' stage, or DNA ploidy status. This study demonstrates that abnormalities of ras genes are a common finding in colorectal carcinomas. They are potentially important biologic changes associated with malignancy, although they do not appear to be related to clinical behavior.

Ras oncogene and the acquisition of metastasizing properties by rectal adenocarcinoma.

To gain a better understanding of the biologic development of rectal adenocarcinomas, the authors evaluated the level of ras gene protein product (p21) in the available material of 74 Dukes' B adenocarcinomas, 64 Dukes' C adenocarcinomas, and 60 lymph-node metastases resected at the University of Chicago Medical Center between 1965 and 1981. Pathologic slides and archival paraffin blocks were retrieved for confirmation of the original diagnosis and measurement of p21 content. P21 titers were obtained using the RAP-5 monoclonal antibody in a semiquantitative immunohistochemical assay. Titer was expressed as the highest dilution giving definitive staining using the avidin-biotin peroxidase method. The analysis indicated that a higher percentage of Dukes' stage C rectal adenocarcinomas had high (greater than or equal to 1:40,000) p21 titers than Dukes' B adenocarcinomas (68.8 vs. 51.4 percent, respectively, P less than 0.05). In view of recent data suggesting that ras oncogene expression confers invasive and metastatic capabilities to NIH 3T3 cells, the authors believe this study offers evidence that overexpression of ras oncogene with overproduction of p21 protein product may be an important prerequisite for the acquisition of metastatic capabilities in the early stages of colon cancer.

[Mucin histochemical changes in various colo-rectal polyps in relation to carcinoma of the large bowel].

210 various colo-rectal polyps including 46 inflammatory polyps, 21 juvenile polyps, 9 hyperplastic polyps, 65 tubular adenomas, 51 familial polyps, 11 villous adenomas, 7 adenomatous polyps with focal cancer, and 14 carcinoma of the large bowel were investigated by HE,HID-AB,PAT-KOH-PAS staining in order to study the mucin changes of these lesions. N-acetylated and C7,C9 O-acetylated sialomucin were mainly obtained in those adenomas with moderate and severe dysplasia (55-64.3%) and the proportion was even higher in cases of villous adenomas, familial polyps, adenomas with focal cancer and advanced carcinoma. These mucins might be assumed as a criteria in representing malignant transformation.