Refinement of an Established Procedure and Its Application for Identification of Hypoxia in Prostate Cancer Xenografts

Cancers

Morten Busk¹  Lars Dyrskjøt²  Trine Strandgaard²  MathildeBorg Houlberg Thomsen²  Steffen Nielsen³  PernilleB. Elming³  BritaS. Sørensen³  MichaelR. Horsman³  ThomasR. Wittenborn³ 

[1] Danish Center for Particle Therapy, Aarhus University Hospital, 8200 Aarhus, Denmark;Department of Molecular Medicine, Aarhus University Hospital, 8200 Aarhus, Denmark;Experimental Clinical Oncology-Department of Oncology, Aarhus University Hospital, 8200 Aarhus, Denmark;

关键词: hypoxia;    pre-clinical models;    prostate cancer;    PC3;    DU-145;    hypoxia gene signature;   

DOI  :  10.3390/cancers13112602

来源: DOAJ

【 摘 要 】

Background: This pre-clinical study was designed to refine a dissection method for validating the use of a 15-gene hypoxia classifier, which was previously established for head and neck squamous cell carcinoma (HNSCC) patients, to identify hypoxia in prostate cancer. Methods: PC3 and DU-145 adenocarcinoma cells, in vitro, were gassed with various oxygen concentrations (0–21%) for 24 h, followed by real-time PCR. Xenografts were established in vivo, and the mice were injected with the hypoxic markers [18F]-FAZA and pimonidazole. Subsequently, tumors were excised, frozen, cryo-sectioned, and analyzed using autoradiography ([18F]-FAZA) and immunohistochemistry (pimonidazole); the autoradiograms used as templates for laser capture microdissection of hypoxic and non-hypoxic areas, which were lysed, and real-time PCR was performed. Results: In vitro, all 15 genes were increasingly up-regulated as oxygen concentrations decreased. With the xenografts, all 15 genes were up-regulated in the hypoxic compared to non-hypoxic areas for both cell lines, although this effect was greater in the DU-145. Conclusions: We have developed a combined autoradiographic/laser-guided microdissection method with broad applicability. Using this approach on fresh frozen tumor material, thereby minimizing the degree of RNA degradation, we showed that the 15-gene hypoxia gene classifier developed in HNSCC may be applicable for adenocarcinomas such as prostate cancer.

【 授权许可】

Unknown