Breast cancer after curative chemotherapy in non-Hodgkin's lymphoma: examination of the role of drug resistance and retrospective comparison to the outcome of de novo breast cancer.

We examined the outcome of patients who developed breast cancer after curative chemotherapy (CHOP) for aggressive non-Hodgkin's lymphoma (NHL) in comparison to the outcome of a retrospectively selected matched-pair group of patients with de novo breast cancer, and evaluated the role of drug resistance-related protein (MDR, MRP, LRP) expression in breast cancer tissue. Twenty-two patients presented with breast cancer (BC) in complete remission after CHOP for NHL. The median age was 62 (49-70) years, each had high/intermediate grade B-cell NHL treated with 6 courses of CHOP, and were in complete remission. These patients were compared to a matched-pair group of de novo BC patients selected from our database over the same time period. Breast cancer tissue was stained by immunohistochemistry for drug resistance proteins LRP, MRP, and MDR. Breast cancer developed after a median of 26 (9-49) months of NHL diagnosis; breast tumor grades 1-2 were seen in 12, and grade 3 in 10 patients; 15 were negative and 7 weakly positive for estrogen and progesterone receptors. Twelve patients were stage IIIA/B, and 10 stage IV and were treated with conventional chemotherapy regimens. All progressed early in liver (n=13), brain (n=9), lung (n=6), bone (n=8), lymph nodes (n=7) and soft tissue (n=5), and received second-line chemotherapy with mitomycin-C + vinblastine or taxanes. The overall survival was 11.8 (6-26) months (p<0.01). Time from NHL to breast cancer development was 19 (14-27) months in patients with positive drug resistance proteins (group A), and 37 (26-56) months in patients with 1 or 2 positive resistance proteins (group B) (p<0.001). In patients with stage IIIA/B disease, there was no difference between the examined and control matched-pair group in median TTP, but there was in overall survival (OS) (23 vs 36 months, p=0.029). In advanced disease, there were more responders in the control vs the examined group (p=0.07). Patients in the control matched-pair group had more prolonged OS when compared to group A patients who developed BC in <24 months from NHL to BC (p=0.017). We conclude that breast cancer developing shortly after a complete response in NHL, is an aggressive disease variant with minimal potential for response to conventional chemotherapy. Analysis of drug resistance mechanisms concerning MDR, MRP and LRP indicates that most of these patients have BC that overexpress these proteins leading to the suggestion that these mechanisms might be a part of the aggressive disease phenotype and partially explain the poor outcome.

Markers of drug resistance in relapsing colon cancer.

PURPOSE: 5-Fluorouracil failure and drug resistance, which often occurs during chemotherapy, is still a great obstacle to the success of human colon cancer treatment. Thus, the comparative study of markers of drug resistance in cancer cells before and after chemotherapy may be extremely helpful in the selection of the appropriate chemotherapeutic drug in colon cancer patients who fail adjuvant treatment with 5-fluorouracil. In the present study we examined the differential expression of three multidrug resistance-related proteins (i.e., P-glycoprotein, MRP, and LRP) and of topoisomerase IIalpha in a series of 20 primary colon carcinomas and their recurrences. METHODS: All markers were determined at tissue level by three-step immunohistochemistry using appropriate monoclonal antibodies, and the markers' immunopositivity was quantified by image analysis. In addition, Feulgen stain was used for the assessment of nuclear DNA content of malignant cells at their primary location. RESULTS: Some degree of aneuploidy was detected in all primary carcinomas. The immunoexpression of the three multidrug resistance-related proteins did not change significantly, either qualitatively (positivity vs negativity) or quantitatively, after chemotherapy. On the contrary, the percentages of topoisomerase IIalpha-positive malignant cells were significantly increased in the tumour recurrences by comparison to their primary locations ( P=0.011). CONCLUSIONS: According to our results, increased topoisomerase IIalpha immunohistochemical expression appears to be part of the malignant cells' phenotype in recurrent colon cancers. Therapeutic options after failure of 5-fluorouracil-based treatment could therefore include appropriate topoisomerase IIalpha-targeted drugs.

Immunodetection and clinico-pathological correlates of two tumour growth regulators in laryngeal carcinoma.

Activation of telomerase, present in the vast majority of all human cancers, is associated with elongation of chromosomal telomeres and consequent cell immortalization. Telomere length homeostasis is a dynamic process governed by the negative feedback mechanism of the telomeric repeat binding factor 1 (TRF1) which inhibits the action of telomerase in telomerase-positive cells. In an attempt to investigate markers of tumour growth as possible prognostic indicators in laryngeal cancer, we studied the expression of TRF1 and of the proliferation marker Ki67 on 96 invasive squamous carcinomas of the larynx. A standard three step immunoperoxidase staining method was applied on paraffin sections incubated with appropriate polyclonal antibodies. The percentages of Ki67- and TRF1-immunopositive cancerous cells were calculated by image analysis. Univariate and multivariate statistical analysis of the staining results were performed in order to detect any association of the examined immunomarkers with the tumours' classical clinicopathological variables including nuclear morphometric features as well as with patients' disease-free survival. Ki67 immunostaining was positively linked with advanced patients' age, nodal involvement as well as presence of early recurrence. No relation was found between proliferative fraction and TRF1 immunoexpression. TRF1 was expressed in 55.2% of all cases and was positively linked only to tumour size. Multivariate statistical analysis revealed the presence of lymph nodal metastasis and Ki67 immunopositivity index > or = 20% as significant predictors of relapse. Increased Ki67 immunostaining appears to be a promising marker of tumour aggressiveness in laryngeal cancer. After one point at the tumour's natural history, the maintenance of tumour growth does not seem to depend on cell proliferation but on TRF1 immunoexpression. Whether the latter can be used for the identification of immortalized cells in every-day practice is worth investigating.

Evaluation of DNA topoisomerase IIalpha expression provides independent prognostic information in non-Hodgkin's lymphomas.

AIMS: In view of the dual role that DNA topoisomerase IIa (TopoIIa) plays as a cell proliferation marker and as a possible indicator of chemosensitivity, we investigated its expression in non-Hodgkin's lymphomas (NHL) in relation to conventional clinicopathological parameters, cell proliferation (as defined by Ki67 immunoreactivity), response to therapy and patient outcome. METHODS AND RESULTS: Formalin-fixed paraffin-embedded tissues from 153 patients with NHL were immunohistochemically stained for TopoIIalpha. Patients were followed up until death (n = 63) or for an average of 68 months (median 64 months, n = 90). The percentage of TopoIIalpha positive cells (TopoIIalpha LI) increased with grade (P < 0.001), extranodal location (P = 0.05) and Ki67 LI (P = 0.01, r = 0.673). In most cases (58%), Ki67 LI exceeded TopoIIalpha LI (TopoIIalpha/Ki67 < 1), especially within the indolent group (P < 0.001). TopoIIalphaLI, Ki67LI and TopoIIalpha/Ki67 ratio were all adversely related to overall survival in univariate analysis, though their significance was not maintained after adjustment for grade. In multivariate analysis high TopoIIalpha/Ki67 ratio and high TopoIIalpha LI independently predicted shortened overall and post-relapse survival, respectively. Most importantly, low TopoIIalpha/Ki67 ratio was the only independent predictor of diminished disease-free survival. However, there was no relationship between TopoIIalpha expression and response. CONCLUSIONS: Our results suggest that evaluation of TopoIIalpha expression and TopoIIalpha/Ki67 ratio as cell proliferation markers provides independent prognostic information in relation to post-relapse and overall survival. Furthermore, TopoIIalpha/Ki67 ratio appears to play a key role in the identification of patients prone to early relapse.

Expression of a cell surface antigen recognized by the monoclonal antibody AUA1 in bladder carcinoma: an immunohistochemical study.

The expression of a cell surface antigen recognized by the epithelium-specific, tumor-associated monoclonal antibody (Moab) AUA1 has been studied in 22 cases of normal urothelium, 7 cases of dysplastic urothelium and 86 cases of transitional cell carcinoma. Tissue specimens were stained with the conventional hematoxylin-eosin technique as well as with the indirect immunoperoxidase technique using Moab AUA1. Results showed that: (a) Moab AUA1 reacts mainly with cells of high or intermediate grade transitional cell carcinoma and with a restricted number of normal urothelial cells, and (b) antigenic heterogeneity exists between tumors of the same grade and within the same tumor.