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Comparison between hypoxic markers pimonidazole and glucose transporter 1 (Glut-1) in murine fibrosarcoma tumours after electrochemotherapy

Comparison between hypoxic markers pimonidazole and glucose transporter 1 (Glut-1) in murine fibrosarcoma tumours after electrochemotherapy Background. Tumour hypoxia occurs as a result of an inadequate supply of blood-borne oxygen due to the disorganized and chaotic vascular network that develops in tumours. Because tumour hypoxia has been associated with a more aggressive phenotype and lower cure rate, there is a recognized need for a method of measuring tumour hypoxia that is suitable for widespread clinical use. The aim of the current study was to compare the expression of Glut-1 with the binding of the bioreductive hypoxia marker pimonidazole and to elucidate the characteristics and pitfalls when they are used as hypoxic markers. Materials and methods. In the study, SA-1 solid subcutaneous tumours in A/J mice were treated by bleomycin given i.v. (1 mg/kg), or the application of electric pulses (8 pulses, 1400 V, 100 μs, 1Hz), or a combination of the two - electrochemotherapy. Pimonidazole was injected 16 hours before tumour excision. Tumours were excised at different time points (0.5, 1, 2, 8, 14 and 24 hours) after therapy. Immunohistochemistry for Glut-1 and pimonidazole adduct was carried out on two consecutive tumour sections and the percentages of positive staining areas were determined. Results. Glut-1 staining was membranous and typically expressed peri-necrotically, whereas pimonidazole staining, although showing substantial co-localisation with Glut-1, was cytoplasmatic. More than 65% of the stained areas showed a high degree of colocalization when the two markers were compared. Our results show that Glut-1 expression significantly correlates with the level of pimonidazole binding (r = 0.41; p = 0.028). Conclusions. Our study confirms that HIF-1 regulated genes, such as Glut-1, have potential for future use as predictors of a decreased sensitivity of tumours to radio- and chemotherapy mediated by hypoxia.