Hanne Havsteen

Measurements of hypoxia using pimonidazole and polarographic oxygen-sensitive electrodes in human cervix carcinomas.

BACKGROUND AND PURPOSE The measurement of tumour oxygenation using Eppendorf oxygen-sensitive needle electrodes can provide prognostic information but the method is limited to accessible tumours that are suitable for electrode insertion. In this paper the aim was to study the relationship between such physiological measurements of tumour hypoxia and the labelling of tumours with the hypoxia-specific marker pimonidazole. MATERIALS AND METHODS Assessment of tumour oxygen partial pressure (pO(2)) using an Eppendorf pO(2) histograph and immunohistochemical pimonidazole labelling was carried out in 86 patients with primary cervix carcinomas. Pimonidazole was given as a single injection (0.5 g/m(2) i.v.) and 10-24 h later pO(2) measurements were made and biopsies taken. Tumour oxygenation status was evaluated as the median tumour pO(2) and the fraction of pO(2) values </=10 mmHg (HP(10)), </=5 mmHg (HP(5)) and </=2.5 mmHg (HP(2.5)). Hypoxia was detected by immunohistochemistry using monoclonal antibodies directed against reductively activated pimonidazole. Pimonidazole binding was scored using a light microscope. Each tumour was evaluated by the relative area pimonidazole at highest score and the accumulated area of pimonidazole labelling from score 1 to 4. Necrosis was measured in HE stained sections. RESULTS AND CONCLUSIONS The degree of hypoxia assessed by either pimonidazole binding or invasive electrode measurements varied significantly between tumours. There was a trend that the most hypoxic tumours measured by oxygen electrodes had the highest score of necrosis, and no or little pimonidazole binding. However, this observation was not consistent and there was no correlation between pimonidazole staining expressed in this way and oxygen electrode measurements of hypoxia.

Invasive oxygen measurements and pimonidazole labeling in human cervix carcinoma

PURPOSE This study was designed to compare tumor hypoxia assessed by invasive O2 sensitive electrodes and pimonidazole labeling in primary human cervix carcinomas. METHODS AND MATERIALS Twenty-eight patients with primary cervix carcinomas (FIGO Stage Ib-IVa) were investigated. Both invasive pO2 measurements and pimonidazole labeling were obtained in all patients. Before treatment, patients were given pimonidazole as a single injection (0.5 g/m2 i.v.). Ten to 24 h later, oxygenation measurements were done by Eppendorf histography, and after this procedure biopsies were taken for pimonidazole-binding analysis. Tumor oxygen partial pressure (pO2) was evaluated as the median tumor pO2 and the fraction of pO2 values < or = 10 mmHg (HF10). Biopsies were formalin fixed and paraffin embedded, and hypoxia was detected by immunohistochemistry using monoclonal antibodies directed against reductively activated pimonidazole. Pimonidazole binding was evaluated by a semiquantitative scoring system. RESULTS Both Eppendorf measurements and pimonidazole binding showed large intra-and intertumor variability. A comparison between pimonidazole binding expressed as the fraction of fields at the highest score and HF10 showed a trend for the most well-oxygenated tumors having a low fraction of fields; however, the correlation did not reach statistical significance (p = 0.43, r = 0.165; Spearmans rank correlation test). CONCLUSION Hypoxia measured in human uterine cervix carcinomas is heterogeneously expressed both within and between tumors when assessed by either invasive pO2 measurements or pimonidazole binding. Despite a trend that tumors with high pO2 values expressed less pimonidazole binding, no correlation was seen between the two assays in this preliminary report.

Treatment outcome following radiotherapy in elderly patients with bladder cancer

BACKGROUND AND PURPOSE The optimal treatment of elderly patients with bladder cancer is not established. This study aimed to evaluate prognostic variables for survival and morbidity, which may be important for treatment strategy. MATERIAL AND METHODS The medical records of 94 patients aged > or = 75 years receiving curatively intended radiotherapy for bladder cancer were reviewed retrospectively. RESULTS Median age was 78 years (range 75-93 years). Fifty patients had T1-2 tumors, and 42 patients had T3-4 tumors. The total planned dose was 57.6-62.6 Gy in 24-30 fractions in 6 weeks. In 76 patients, a 2 week rest period was planned after 16 fractions (split course). Half of the patients were hospitalized during or after the treatment because of gastrointestinal or urogenital side effects. Median survival was 13.9 months (range 0.6-150.0 + months), 29% survived for 2 years and 7% survived for 5 years. Patients aged > 78 years survived for a shorter period than patients aged 75-78 years (13.4 versus 16.1 months). Univariate survival analysis revealed that low stage (T1-2), good performance status (PS < or = 1), split course treatment, no treatment interruption due to side effects, and no hospitalization during treatment were associated with long survival. In multivariate analyses, T-stage, split course treatment, and performance status were independent prognostic factors. CONCLUSION The results confirm that curative intended radiotherapy is feasible in elderly patients, but patients with stage T3-4 and PS > 1 have a short survival. These patients should be offered palliative treatment.

Altretamine (hexamethylmelamine) in the treatment of platinum-resistant ovarian cancer: a phase II study

OBJECTIVE To evaluate the activity of oral Altretamine in women with epithelial ovarian carcinoma who responded (PR or CR) to first line chemotherapy but relapsed within 6 months. The protocol was later amended to include patients with relapse within 12 months. METHODS A multicentric phase II trial. The patients had to have measurable disease. No more than one prior chemotherapy regiment was allowed. The patients were treated with 260 mg/m(2)/day of Altretamine in four divided doses for 2 weeks, repeated every 4 weeks. The response was evaluated after every two courses. RESULTS Thirty-one eligible patients were treated with a median of 3 courses of Altretamine (range 1-12). Hematological toxicity was minimal. Gastrointestinal toxicity was common. Response evaluation was possible for 26 patients. Three patients (9.7% intent-to-treat) achieved a partial response. Eight patients had stable disease, and 15 patients had progressive disease after two treatment courses. The median time to progression was 10 weeks (range, 5-51 weeks). Medial survival was 34 weeks (range, 7-112+). CONCLUSION Altretamine should not be chosen as standard treatment in patients with platinum-resistant recurrent ovarian cancer. However, Altretamine represents a useful alternative in patients who prefer oral treatment or when socioeconomic considerations are an important issue.

Fibroblast growth in the soft agar clonogenic assay for cervix cancer radiosensitivity.

Sir, Recently, West and colleagues published a paper in this journal summing up their work on radiosensitivity testing in 128 cervix cancer patients, with a follow-up time from 2 to 5 years (West et al, 1997). The purpose of that study was to test the hypothesis that tumour cell in vitro radiosensitivity measured before treatment predicts clinical outcome of the individual patients after curative radiotherapy alone (Davidson et al, 1990). Tumour biopsies were obtained before treatment, and cellular in vitro radiosensitivity was assessed, using the modified Courtenay-Mills soft agar clonogenic assay, by measuring the fraction of cells surviving after a radiation dose of 2 Gy (SF2). The study concluded that SF2 was a significant prognostic parameter for overall survival, local control and metastasis-free survival, and that this was independent of disease stage, tumour grade, patient age, colony-forming efficiency and tumour diameter. The biological explanation for this finding is still not clear-cut, as we now demonstrate, that when culturing biopsies from carcinoma of the uterine cervix, both stromal fibroblasts as well as tumour cells can be grown. Our approach for culturing biopsies from carcinoma of the uterine cervix involves removing the soft agar, and all the colonies are collected on preparation slides for identification of the origin of those colonies. For immunocytochemistry, the monoclonal antibodies anti-cytokeratin (AE1-3, Biogenex), reacting with epithelial cells, and anti-vimentin (3B4, Dako), reacting with fibroblasts, are used (Stausb0l-Gr0n et al, 1995; 1998). Twelve carcinomas of the uterine cervix (nine squamous cell carcinomas, two adenocarcinomas, one adenosquamous carcinoma) met the criteria for successful growth, with more than ten colonies in the unirradiated tubes. Plating efficiency, irrespective of cell type ranged from 0.004% to 0.297% with a median of 0.02 1%, concordant with the results reported by West et al ( 1997). The minority of the colonies in the unirradiated cultures of most cervix carcinoma biopsies was tumour marker positive, ranging from 0% to 93%, with a median of 26% (Figure 1). In parallel, the unirradiated tubes contained 6-100% fibroblast marker-positive colonies, with a median of 80%. The sum ranged from 80% to 125%. Cellular in vitro radiosensitivities of tumour cells (tumour cell SF2), fibroblasts (fibroblast SF2) and an overall estimate (overall SF,) were determined. In ten patients, overall SF2 ranged from 0.31 to 0.81, with a median of 0.57. Tumour cell SF2 and fibroblast SF, had median values of 0.53 (range 0.26-0.67) and 0.55 (range 0.28-1.00) respectively. Previous studies on other tumour types support the finding that primary tumour biopsies are a source of fibroblast colonies when grown in the modified Courtenay-Mills soft agar clonogenic assay (Lawton et al, 1994; Stausb0l-Gr0n et al, 1995). In head and neck carcinomas, the majority of the colonies obtained in the unirradiated tubes originated from fibroblasts, and the overall SF2 was statistically significantly correlated to an independent measure of fibroblast SF, (Stausb0l-Gr0n et al, 1995). Thus, taken together, it may be likely that the radiosensitivity of stromal fibroblasts, dominating the A

Recurrent thromboembolism in ovarian cancer

Thrombosis is a well-known complication in cancer. Recurrent thromboembolism poses a clinical obstacle, as new thromboembolic episodes often occur despite antithrombotic therapy. We present a case of recurrent thrombosis in ovarian cancer. Interestingly, new thromboembolic episodes occurred repeatedly after discontinuing low molecular weight heparin (LMWH) therapy, and conversely, no thrombotic episodes were noted during LMWH therapy.