Lectins and immunohistochemistry of colorectal cancer, its recurrences and metastases.

In 31 patients resected specimens from primary colorectal cancers, corresponding liver metastases and local recurrences were investigated for the staining pattern of lectins (PNL, UEA, WGA, HPA, SBA, RCA) and tissue antigens (CEA, SP, ACT) by immunohistochemistry. Comparison of staining patterns showed a loss of marker expression from normal colonic mucosa to colorectal primary carcinomas, and a tendency to marker loss from the primary tumour to liver metastases. However, even a neo-expression of markers not present in the primary tumour could be observed. For clinical use, serum markers observed in patient follow-up may be valuable even where the findings are negative at the time of primary tumour surgery. In contrast to the heterogenous marker map of primary tumours and metastases, comparison of primary and locally recurrent tumour revealed a staining pattern that was almost always identical. This supports the hypothesis that locoregional recurrences develop from remnant cells of the primary tumour left behind at surgery. There is no support for the thesis that locoregional recurrences arise from mucosal changes at the anastomosis or from suture material.

Flow cytometric DNA ploidy analysis of feline mammary tumors.

Flow cytometric DNA analysis was performed on biopsies from 9 nonmalignant and 111 malignant (primary and metastatic) feline mammary lesions. In our series, 46.3% of the primary mammary carcinomas appeared to be aneuploid, whereas all but one benign breast lesion were diploid. The degree of aneuploidy in carcinomas was low, with a relatively high number of primary tumors (12 of 82) displaying hypodiploidy. Aneuploidy was not found to be correlated with any specific histological tumor type, vascular invasion, tumor size, or histological malignancy grade or with the separate components thereof. Comparison of the ploidy in primary and metastatic tumors from the same cases revealed a remarkable stability, both in time and location of appearance of the metastases. It is concluded that with respect to DNA ploidy feline mammary carcinoma has more in common with canine mammary carcinoma than with human mammary carcinoma. Further prospective studies are necessary to clarify the implications of aneuploidy in feline mammary carcinoma for tumor behavior and prognosis.

DNA ploidy in endometrial carcinoma: major objective prognostic factor.

Paraffin-embedded tissue samples from 256 patients who received primary treatment (surgical staging, reduction of tumor size, and adjuvant therapy based on surgical and pathologic risk factors) for endometrial carcinoma at the Mayo Clinic between 1979 and 1983 were analyzed by flow cytometry to determine DNA ploidy characteristics. Diploid patterns constituted 78% of the cases, whereas aneuploid and tetraploid patterns accounted for 17% and 5%, respectively. Only 10% of patients with diploid tumors had a relapse in comparison with 39% of those with nondiploid lesions (34% with aneuploid; 58% with tetraploid). Significant differences (P less than 0.001) were noted in estimated 4-year progression-free survivals--88% for patients with diploid and 57% for those with nondiploid tumors. Stage, grade, depth of myometrial invasion, histologic subtype, peritoneal cytology, and DNA ploidy all demonstrated independent prognostic significance (P less than 0.001) in this study population. When subjected to multivariate analysis, however, grade and depth of myometrial penetration failed to retain prognostic significance (P greater than 0.15) and surgical stage was marginally significant (P = 0.05), whereas histologic subtype and DNA ploidy maintained significant predictive powers (P less than 0.001 and P less than 0.01, respectively). We conclude that DNA ploidy is a major objective prognostic factor and therapeutic determinant for endometrial carcinoma.

Expression of epidermal growth factor receptor in breast carcinoma.

A series of 90 patients with primary operable breast cancer and with up to 36 months of follow up were investigated for expression of epidermal growth factor receptor (EGFR) in their tumours by immunocytochemical staining with the monoclonal antibody EGFR 1. Tumour samples were snap frozen in liquid nitrogen immediately after resection and subsequently stained using a standard indirect immunocytochemical method. Tumour staining was assessed by two observers and scored on a four point scale (0-3). Thirteen (14%) tumours showed positive immunoreactivity. A strong correlation between distinct EGFR expression and short disease free interval was observed. Significant correlations were also shown with oestrogen and progesterone receptor expression and tumour nuclear size. No significant association was found with tumour size, lymph node stage, and histological grade. The association with disease free interval remained significant in multivariate analysis.

Association between poor prognosis in early-stage invasive cervical carcinomas and non-detection of HPV DNA.

Human papilloma virus (HPV) DNA sequences (HPV types 16, 18, 33, 35 or uncharacterized) were detected by Southern blot hybridisation and polymerase chain reaction in 84% of 106 early-stage invasive carcinomas of the uterine cervix. Among HPV-positive patients, the risk of overall relapse did not differ with individual HPV types. Compared with HPV-positive patients, those with no detectable HPV DNA had a 2.6 times higher risk of overall relapse (p less than 0.05) and 4.5 times higher risk of distant metastases (p less than 0.01). The 24-month relapse-free survival rate in HPV-positive patients was significantly higher than that in HPV-negative patients (77% vs 40%), and the difference was similar (91% vs 56%) among those who were node-negative. These data indicate that HPV-negative cervical carcinomas may represent a biologically distinct subset of tumours that carry a poorer prognosis than do HPV-positive cancers.

[Flow cytometric DNA analysis of papillary carcinoma of the thyroid using paraffin-embedded specimens].

A flow cytometric DNA analysis of papillary thyroid carcinomas has been performed on paraffin-embedded specimens, in order to determine the clinical usefulness of this method by defining the degree of the biological malignancy. Fifty-eight patients with papillary carcinomas were followed from 2 to 8 years. They were classified according to their prognosis into three groups: non-recurrent (47 patients), recurrent (5 patients), and a deceased group (6 patients). Aneuploidy was found in 6 (10.3%) of the total 58 patients. It was more frequently detected in the recurrent (40%) and in the deceased (33.3%) groups than in the non-recurrent group (4.3%). Higher proliferating index values were observed in the deceased group than in the recurrent and non-recurrent groups. These results indicate that a DNA analysis, using routine paraffin-embedded materials, is a useful adjunct in the determination of the degree of the biological malignancy of a papillary carcinoma.

[Relationship between the effectiveness of CDDP therapy and the nuclear DNA content in advanced-recurrent gastric cancer cases].

A microspectrophotometric analysis of the DNA content has been performed on 9 advanced recurrent gastric cancer patients with measurable lesions, who has either been treated by CDDP alone or with other chemotherapeutics during a three-year period since, 1984. Histograms of the DNA content were classified into four ploidy patterns. All of the 4 responder cases (CR, PR, MR) showed type, IV, although only one of 5 non-responder cases revealed the same typing. In one of the two CR cases the DNA ploidy pattern, which was examined before and after the therapy, changed from type IV to type II. Thus it appears that an analysis of the DNA content may be useful in evaluating the effectiveness of different chemotherapies.

Primary recurrent leiomyosarcoma of the breast. Case report with ultrastructural and immunohistochemical study and review of the literature.

The authors report a case of recurrent mammary leiomyosarcoma in a 50-year-old woman. The neoplasia, with a recognized clinical evolution of 11 years, was resected on two occasions and had not metastasized. Microscopic examination showed 4 mitoses/10 high-power fields, moderate cytologic atypia, and, ultrastructurally, abundant myofibrils with condensations. Immunoperoxidase stains had positive results for muscle-specific antigen and showed focal reactivity for epithelial membrane antigen and S-100 protein. Analysis of the ten cases (including the present one) reveals that this neoplasm has appeared with greater frequency in women with an average age of 52 years. All neoplasms have been limited to the breast at the time of diagnosis. As a group, they have better prognosis than other sarcomas of the breast, although the possibilities of recurrence or dissemination exist, even many years after the primary extirpation. The size of the tumor and mitotic activity seem to be of little prognostic value. Mammary leiomyosarcoma shares clinical and pathologic similarities with subcutaneous leiomyosarcoma in other anatomic sites.

[Regrowth patterns of glioma--cases of glioma regrew away from the original tumor].

There have been conflicting opinions regarding the correct volume to be used in radiotherapy fields for malignant glioma. Whereas many clinicians recommend limited fields designed to cover the tumor volume with a margin according to the facts about 90% of recurrent malignant glioma regrew within 2 cm of the original tumor and regrowth away from the original tumor was seen in about 5%. Two cases of malignant glioma regrew away for the original tumor and one case of malignant glioma regrew into the resected site were presented. And the biological characteristics of glioma cells concerning with the invasiveness were discussed.

Variations in steroid receptor status with disease stage in breast cancer.

Oestrogen and progesterone receptor (ER and PgR) distribution in three clinical subgroups of 421 breast carcinomas was analysed. The groups comprised (1) early breast cancer (T1-2a, N0M0; n = 64); (2) untreated advanced fungating cancer (n = 27) and (3) advanced cancer relapsing after endocrine therapy (n = 29). Receptor distribution in each of the subgroups was compared to that of the total population. The advanced fungating group contained no ER--ve/PgR--ve tumours and the distribution was also significantly different from the total population (P less than 0.001 by Chi-squared test). The proportion of tumours in the total population that contained greater than 40 fmol/mg ER was 187/421 (44.4%). There was no significant difference between the early breast cancer group and the total population (P greater than 0.9). However, the proportion of tumours containing ER greater than 40 fmol/mg in the advanced fungating cancer group (16/27, 59.3%) was significantly higher than in the total population (P less than 0.01). This difference may be partially explained by the older age at presentation in this group. In the relapsed after endocrine therapy group only four of 29 (13.8%) contained ER greater than 40 fmol/mg which was significantly different from the total (P less than 0.001). There was a higher proportion of early breast cancers containing PgR greater than 40 fmol/mg than in the total population (P less than 0.001). There was no significant difference between PgR distribution in the advanced fungating and relapsed groups compared to the total population. The data suggest that patients presenting with advanced fungating cancer are more likely to respond to endocrine therapy than the population as a whole, and that in breast cancer that has relapsed following endocrine therapy receptor levels decrease with progression of the disease.

Prognostic contributions of Helix pomatia and carcinoembryonic antigen staining using histochemical techniques in breast carcinomas.

Helix Pomatia (HPA) and carcinoembryonic antigen (CEA) staining were studied in 113 primary breast carcinomas, 63 metastatic lymph nodes and 10 resected local recurrences, using the avidin-biotin-peroxidase complex (ABC) method. Positive percentage rates were 41 (46/113) for HPA and 24 (27/113) for CEA in primary tumors, and 80 (8/10) for HPA but only 10 (1/10) for CEA in metastatic lesions. HPA staining showed a statistically significant correlation with negative estrogen receptor (ER) status (r -0.25, P = 0.05) and high nuclear grade (r 0.20, P = 0.04). Cancers with positive HPA staining were associated with a lower survival rate than those without (P = 0.0001), irrespective of menopausal status. CEA staining showed a marginal correlation with survival (P = 0.06: log rank test). The five-year survival rate of 21 cases positive for these two markers was only 5%. Multivariate analysis revealed HPA staining to be the best prognostic factor. The data indicated that HPA staining might be a valuable prognostic factor for breast cancer patients, especially when combined with CEA staining.

Minimal residual disease in acute lymphoblastic leukemia detected by immune selection and gene rearrangement analysis.

We have developed an assay for the detection of malignant residual cells in the bone marrow from patients with B- or T-lineage acute lymphoblastic leukemia (ALL) in clinical remission. This assay involves an immune selection step followed by immunoglobulin or T-cell receptor gene rearrangement analysis and allows the detection of one contaminating tumor cell out of 1,000 normal bone marrow cells. We have examined the bone marrow of 11 patients with adult ALL in remission over a 24-month period. Five patients relapsed in the bone marrow and one in the CNS. The assay allowed the detection of minimal residual disease in four of five patients that subsequently relapsed in the bone marrow, 1.5 to 9 months before the relapse became morphologically and clinically manifest. Residual disease was not found in the bone marrow from patients in continuous remission and from the single patient who relapsed in the CNS. We conclude that the ability of the assay described here to detect minimal residual disease with high specificity can provide information for further understanding of the biology of ALL and hopefully for the clinical management of patients with this disease.

Breast cancer, prostaglandins and patient survival.

Prostaglandins may have both undesirable and desirable effects in malignant disease. Their possible roles in breast cancer were studied by examining the relationships between different variables and the amounts of prostaglandin-like material (PG-LM) extracted from 141 breast carcinomas. Univariate analysis indicates a direct correlation with patient age and menopausal status, with a greater yield from cancers of post- compared with pre-menopausal women. Tumours up to 2 cm diameter yielded more PG-LM than those measuring greater than 2-5 cm. Although there was also a direct correlation with bone metastasis near to the time of surgery, this was because no positive bone scans occurred in patients whose tumours yielded little total PG-LM (less than 16 ng PGE2 equivalents per g tissue). Since tumour PG-LM did not predict later spread to bone, and yields of greater than 16 ng g-1 were similar in the positive and negative bone scan groups, tumour PG-LM appears to be unimportant for skeletal metastasis. There was no obvious relationship of tumour PG-LM to the grade of malignancy, tumour type, amounts of fibrous tissue (and therefore malignant cells), invasion of blood vessels and lymphatics or presence of plasma cells. Multivariate analysis indicates that disease-free survival is longest with an intermediate production of tumour total PG-LM. Of the 82 patients now dead, the cause was attributed to metastatic disease in 69 cases. No relationship of PG-LM to the length of survival was seen with univariate or multivariate analysis. However, when just the post-menopausal patients who died within the first 3 postoperative years were analysed, there was a highly significant inverse correlation between the tumour total PG-LM and the time to death. The reason(s) for these different findings on overall survival compared with just the patients who died are not understood, but the results may indicate that one or more other variables must co-exist with a high tumour PG-LM to hasten death.

Possible role for vitamin D in controlling breast cancer cell proliferation.

By means of an immunocytochemical method the 1.25-dihydroxyvitamin D [1.25(OH)2D] receptor status of tumours from 136 patients with primary carcinoma of the breast was determined. Patients with receptor-positive tumours had significantly longer disease-free survival than those with receptor-negative tumours (Chi2 = 4.01, p less than 0.05). 1.25(OH)2D3 inhibits the proliferation of several established human breast cancer cell lines in vitro. Effects of 1.25(OH)2D3 on breast tumour growth in vitro were assessed by means of the nitrosomethylurea-induced rat mammary tumour model of hormone-responsive breast cancer. Treatment of tumour-bearing animals with 0.1 microgram of the synthetic analogue, 1 alpha-hydroxyvitamin D3, three times weekly produced significant inhibition of tumour progression. Taken together, these studies suggest that the levels of 1.25(OH)2D occurring in vivo may exert an inhibitory effect on receptor-positive tumours. Further studies are required to evaluate the role of vitamin D metabolites in the treatment of human malignant disease.

Expression of epidermal growth factor receptors associated with lack of response to endocrine therapy in recurrent breast cancer.

Epidermal growth factor receptors (EGFR) and oestrogen receptors (ER) were analysed in 221 patients with primary operable breast cancer by means of radioligand assays. After median follow-up of 24 months (range 3-60 months), there had been recurrences in 99 patients, of whom 72 (median age 56 years, range 32-77 years) received tamoxifen alone as first-line treatment for recurrence. 20 patients (28%) showed a response to this therapy and 52 (72%) did not. Of 32 ER-positive tumours, 12 (37.5%) showed an objective response to tamoxifen compared with only 2 of 40 (5%) ER-negative tumours (p less than 0.005). Of 35 EGFR-positive tumours, 3 (8.5%) achieved an objective response compared with 11 of 37 (30%) EGFR-negative tumours (p less than 0.05). Only 1 of 28 EGFR-positive, ER-negative tumours achieved an objective response. Including patients whose disease remained stable for more than 6 months with the responders, however, EGFR status was a better predictor of response to tamoxifen; 15 of 37 EGFR-negative patients and 5 of 35 EGFR-positive patients responded (p less than 0.01), compared with 13 of 32 ER-positive and 7 of 40 ER-negative patients (not significant). EGFR expression is a highly significant marker of poor prognosis in patients with breast cancer; it appears to be as good a predictor as ER for objective response and better for overall response to endocrine therapy on relapse.

A prospective study of the use of 111In-labelled monoclonal antibody against carcino-embryonic antigen in colorectal cancer and of some biological factors affecting its uptake.

This prospective study of radioimmunoscintigraphy using 111In-labelled, C46, a monoclonal antibody against carcino embryonic antigen, CEA, was undertaken in 23 patients with colorectal cancer, 18 with primary carcinomas, 3 with recurrences and 2 with metastases. A sensitivity of 95% and an accuracy of 91% was found. New observations through the use of two administered doses of antibody and the analysis of surgical specimens showed that for the low dose of antibody (average 0.76 mg) tumour uptake was 1.14 x 10(-2)% of the injected dose per gram, whereas on the higher dose (average 4.67 mg) the average tumour uptake was 6.77% (same units) p less than 0.01. Thus a six-fold increase of antibody caused a six-fold increase in tumour uptake, but no change was seen in the tumour to mucosa ratio 4.8 +/- 0.5 (low dose) and 4.0 +/- 1.0 (high dose). A further observation was that well and moderately differentiated tumours took up about four times more than poorly differentiated tumours (p less than 0.05). It was also found that tumour free lymph nodes had a six-fold greater uptake than tumour involved nodes (p less than 0.05) confirming other work with anti-CEA monoclonal antibody. In conclusion, although biological factors influence its efficacy, 111In labelled C46 anti-CEA is a suitable radiopharmaceutical for the radioimmunoscintigraphy of colorectal cancer.

Condylomatous carcinoma of the vulva with special reference to human papillomavirus DNA.

Nine cases of condylomatous carcinoma (squamous cell carcinoma arising in condyloma acuminatum) of the vulva were studied for their clinical history, histopathology, and presence of human papillomavirus (HPV) DNA. Condylomatous carcinoma occurred primarily in an elderly population with a mean age of 70 years. There was an antecedent history of vulvar condyloma in 77%, with a median of nine months before the documentation of an invasive lesion. The disease had a good prognosis, with few recurrences and no metastasis or deaths from the disease. Human papillomavirus DNA was demonstrated to be present in 55% of these tumors by either filter or in situ hybridization techniques. Both HPV 6 and HPV 16 DNA were identified in an equal number of cases.

DNA analysis in the differential diagnosis of osteosarcoma.

The DNA content of osteosarcomas, and of giant cell tumors, osteoblastomas, aneurysmal bone cysts, and fibrous dysplasias was determined by cytophotometry. Of 158 tumors, 141 were histologically noncontroversial, whereas 17 posed diagnostic difficulties. In the noncontroversial group, all 41 benign tumors had a diploid (normal) DNA content. Ninety-two of 96 high-grade osteosarcomas were hyperploid (increased DNA content). The four analyzed low-grade parosteal osteosarcomas were diploid. Among 17 diagnostically controversial cases, nine were primarily diagnosed and treated as benign. Three of these patients, nevertheless, exhibited a malignant clinical course and two had local recurrence. All five proved to have hyperploid tumors. The four nonrecurrent lesions were diploid. Of eight patients primarily evaluated as malignant, one died and two developed local recurrence. These three patients had hyperploid tumors. Among the five nonrecurrent lesions, two were hyperploid and three diploid. In the diagnostically controversial group, recurrence or death was consistently related to hyperploidy. The present study shows that the vast majority of high-grade osteosarcomas are hyperploid. Benign bone tumors, which may be mixed up histologically with osteosarcoma, are diploid. Routine DNA analysis of primary bone tumors, as an adjunct to histopathologic assessment, can be employed to obtain diagnostic confirmation. In cases presenting histopathologic difficulties, ploidy determination may provide decisive diagnostic information.

DNA analysis in the differential diagnosis of osteosarcoma.

The DNA content of osteosarcomas, and of giant cell tumors, osteoblastomas, aneurysmal bone cysts, and fibrous dysplasias was determined by cytophotometry. Out of 158 tumors, 141 were histologically noncontroversial, whereas 17 posed diagnostic difficulties. In the noncontroversial group all 41 benign tumors had a diploid (normal) DNA content. Ninety-two of 96 high-grade osteosarcomas were hyperploid (increased DNA content). The four analyzed low-grade parosteal osteosarcomas were diploid. Among 17 diagnostically controversial cases, nine were primarily diagnosed and treated as benign. Three of these patients, nevertheless, exhibited a malignant clinical course and two had local recurrence. All five proved to have hyperploid tumors. The four nonrecurrent lesions were diploid. Of eight cases primarily evaluated as malignant, one died and two developed local recurrence. These three patients had hyperploid tumors. Among the five nonrecurrent lesions, two were hyperploid and three diploid. Hence, in the diagnostically controversial group, recurrence or death was consistently related to hyperploidy. The current study shows that the vast majority of high-grade osteosarcomas are hyperploid. Benign bone tumors, which may be mixed up histologically with osteosarcoma, are diploid. Routine DNA analysis of primary bone tumors, as an adjunct to histopathologic assessment, can be employed to obtain diagnostic confirmation. In cases presenting histopathologic difficulties, ploidy determination may provide decisive diagnostic information.