The development of a molecular screening method for cancer patients is of great importance, since it would contribute to the selection of the most effective chemotherapy regimen for each patient. In the present study we applied such a method, semi-quantative RT-PCR analysis, and we examined the expression of the multidrug resistance gene MDR-1, the metastasis suppressor gene nm23-H1 and the non-MDR drug resistant gene H Sema E in 53 ovarian and breast cancer specimens. Moreover, we have correlated the expression profile of these genes with the histopathological findings and clinical outcome of the examined patients. The majority of specimens were found to be positive for MDR-1 and H Sema E gene expression, while nm23-H1 was detected in less than 50% of the patients. Correlation and statistical analysis of the molecular data with clinicopathological features showed that nm23-H1 could serve as a good prognostic factor for ovarian cancer patients. In breast cancer patients, nm23-H1 expression was associated with a 6.1 higher death risk. Ovarian cancer patients who express nm23-H1, but not MDR-1 and H Sema E, tend to have longer survival than patients with any other gene combination. Finally, breast cancer patients with advanced disease showed a better response when they were negative for all the three genes studied. In conclusion this work proposes that the combined study of the expression of different genes may be a useful approach for evaluating patients' response to therapy.
Conflicting results exist regarding the significance of the metastasis suppressor gene nm23-H1 in cancer patients. Initial results from a study done by our group were more indicative of its negative prognostic role in breast cancer. Our aim was to examine further its significance in patients with metastatic breast cancer.With the semi-quantitative Reverse Transcripted-Polymerase Chain Reaction (RT-PCR) method, solid tumor specimens or samples from malignant effusions in breast cancer patients were examined for the nm23-H1 gene. Clinical data were collected retrospectively and gene expression was correlated with survival.Fourteen patients were included in the current analysis. The gene was detected in 7 patients. No statistically significant differences were observed in the comparison done for prognostic factors between nm23-H1-positive and nm23-H1-negative patients. Women in whom the gene was not detected had longer median survival (49 vs. 6 months, p=0.09).In advanced breast cancer, nm23-H1, as detected by RT-PCR, seems to be a predictor of bad prognosis.
Six patients with metastatic breast cancer and malignant pleural effusions and 13 patients with known or suspected ovarian cancer, underwent immunoscintigraphy after intracavitary (intrapleural or intraperitoneal) administration of iodine-131-(131I) or indium-111-(111In) labeled tumor associated monoclonal antibodies HMFG2 and H17E2. This method proved to be sensitive and specific with a true-positive result in 13 out of 14 patients with tumor and a true-negative result in five out of five patients without tumor. At any one time, 65%-80% of the whole-body radioactivity was closely associated with the cavity into which the radiolabeled antibody was administered while the radioactivity in the blood was always low, (approximately 4 X 10(-3) of administered dose/ml of blood). Concentrations of radiolabeled antibody (per gram of tumor tissue) ranged from 0.02%-0.1% of the injected dose in intracavitary tumors, but only 0.002% in a retroperitoneal metastasis. The specificity of this approach was documented in four control patients with benign ovarian cysts and in two patients who were imaged using both specific and nonspecific radiolabeled antibody. We conclude that the intracavitary administration of 131I- or 111In-labeled HMFG2 and H17E2 is a favorable route of administration and offers significant advantages over previously reported intravenous administration for the localization of breast or ovarian metastases confined to the pleural or peritoneal cavities.
The antigenic expression of normal bladder epithelium and transitional carcinomas has been studied using the epithelium-specific, tumour-associated monoclonal antibodies HMFG1 and AUA1. Tissues from 79 cases of bladder carcinoma and 11 cases of non-neoplastic bladder tissues were stained with both the haematoxylin-eosin (H/E) and the indirect two-stage immunoperoxidase methods using the monoclonal antibodies (MAbs) HMFG1 and AUA1 at a concentration of 25 micrograms ml-1. Positive and negative controls were also used. Moreover, 46 urine smears prepared after cytocentrifugation were stained with both the Papanicolaou and the indirect two-stage immunoperoxidase methods. The results showed that HMFG1 reacted with the majority of cases of grade III carcinomas and carcinomas in situ and with a subset only of low-grade (I and II) carcinomas. The pattern of staining showed the following characteristics: (1) the epithelial surface membrane stained both in normal bladder and bladder carcinomas (surface of the papillae), (2) a variant number of cancer cells, increasing with the degree of malignancy, showed membrane and/or cytoplasmic staining, (3) tumours of the same histological grade showed antigenic heterogeneity. The MAb AUA1 was not widely expressed. The immunocytochemical study confirmed the reaction of HMFG1 with a variant number of malignant urothelial cells exfoliated in urine. Their reaction with AUA1 was much more limited. The immunocytochemical staining seemed to be more sensitive in the detection of malignant cells in some cases which had been characterized as negative or suspicious for malignancy by the Papanicolaou examination. The intravesical treatment with chemotherapeutic agents did not seem to influence the antigenic expression of malignant urothelial cells.
The serum CA 125 marker is elevated in 80% of patients with ovarian adenocarcinoma. MDR 1 gene expression has been identified in a variety of tumor types and its expression has been correlated with multidrug resistance. Whether there is a correlation between CA 125 levels and MDR 1 expression has not been sufficiently investigated. Therefore, the aim of this study was to examine whether an association between serum CA 125 levels and MDR 1 expression exists.Serum CA 125 levels were measured during the diagnosis of ovarian cancer. Fresh tumor specimens or ascitic fluid samples were studied for MDR 1 expression by the polymerase chain reaction method (PCR).Forty patients with ovarian cancer were studied, 34 (85%) of whom had elevated CA 125. Twenty-eight out of the 40 patients were tested for MDR 1 expression; 20 expressed the gene and 8 did not. The median level of CA 125 in specimens expressing the MDR1 gene was 327, and in specimens that did not it was 376. There was no correlation between the CA 125 levels and MDR 1 expression (p = 0.484).There does not seem to be an association between CA 125 levels and expression of the MDR1 gene in patients with ovarian cancer.
In a 42 year male, with hepatocellural carcinoma, confirmed by biopsy and a positive for somatostatin receptor liver scan, without any symptoms of neuroendocrine activity, it was decided to be treated with high therapeutic doses of [In-111-DTPA-D-Phe1] Octreotide [Mallinckrodt Br, Petten] and to abandon the conventional and, in the majority of the cases, hopeless cytostatic therapy.
Immunoscintigraphy of ovarian tumors by intraperitoneal administration of I131 HMFG2 monoclonal antibodies (mabs) was used in this study. The purpose was to evaluate the diagnostic potential of this non-operative imaging technique in detecting ovarian tumor nature and spread. Sixteen patients that received 500-1000 microCi of I131 labelled HMFG2 mabs were evaluated. The scans obtained were compared mainly with the macro and microscopic operative findings of the subsequent laparotomy. Immunoscintigraphy accurately scanned tumor spread in 7 out of 9 patients with known ovarian cancer. It also successfully revealed the malignant or benign nature of pelvic masses in 6 out of 7 patients examined.
We have previously shown that metastatic carcinomas of unknown primary site overexpress several tumor markers as well as the products of the oncogenes c-myc, ras and c-erbB2. We analyzed the tissue expression of the protein products of the apoptosis modulation genes p53 and bcl-2 in 47 CUP cases. Formalin-fixed, paraffin embedded tumor specimens were stained with commercially available antibodies to p53 (DO7) and bcl-2 after antigen retrieval by the microwave method. Staining was evaluated by intensity (1+ to 3+), percentage of positive cells (1-100%), and the 'intensity times percentage' product defined as the immunoreactivity index with values ranging from 0 to 300. Immunoreactivity index values higher than 150 were considered to characterize protein over-expression. Expression of p53 was identified in 70.2% of tumors while 53% of them showed a high immunoreactivity index. Bcl-2 expression was detected in 65% of tumors and overexpressed in 40%. Overexpression of both proteins was detected in 20% of tumors. The detection of either protein was not associated with any of the major clinicopathological variables studied. Nevertheless, a trend towards a more favourable response to platin based chemotherapy was seen in the cases that showed a strong expression of both proteins, when analysed by immunoreactivity index and percentage of positive cells. We conclude that CUP overexpress at a high percentage the p53 and the bcl2 proteins. The observed weak association of strong expression of these proteins with response to platin-based chemotherapy deserves further evaluation in the CUP setting.
