Hypoxia biomarkers in squamous cell carcinoma of the uterine cervix

Christine Ellingsen, Lise Mari K Andersen, Kanthi Galappathi, Einar K Rofstad, Christine Ellingsen, Lise Mari K Andersen, Kanthi Galappathi, Einar K Rofstad

Abstract

Background: There is significant evidence that severe tumor hypoxia may cause resistance to chemoradiotherapy and promote metastatic spread in locally advanced carcinoma of the uterine cervix. Some clinical investigations have suggested that high expression of hypoxia-inducible factor-1α (HIF-1α) and/or its target gene carbonic anhydrase IX (CAIX) may be useful biomarkers of tumor hypoxia and poor outcome in cervical cancer. Here, we challenged this view by investigating possible associations between HIF-1α expression, CAIX expression, fraction of hypoxic tissue, and lymph node metastasis in experimental human tumors.

Methods: Tumors of two cervical carcinoma xenograft lines (CK-160 and TS-415) were included in the study. Pimonidazole was used as a hypoxia marker, and tumor hypoxia, HIF-1α expression, and CAIX expression were detected by immunohistochemistry. Metastatic status was assessed by examining external lymph nodes in the inguinal, axillary, interscapular, and submandibular regions and internal lymph nodes in the abdomen and mediastinum.

Results: Tissue regions staining positive for pimonidazole, HIF-1α, or CAIX were poorly colocalized, both in CK-160 and TS-415 tumors. The expression of HIF-1α or CAIX did not correlate with the fraction of hypoxic tissue in any of the two tumor lines. Furthermore, clinically relevant associations between HIF-1α or CAIX expression and lymph node metastasis were not found.

Conclusion: Because significant associations between HIF-1α expression, CAIX expression, fraction of hypoxic tissue, and incidence of lymph node metastases could not be detected in any of two preclinical models of human cervical cancer, it is not realistic to believe that high expression of HIF-1α or CAIX can be useful biomarkers of tumor hypoxia and poor outcome in a highly heterogeneous disease like cervical carcinoma.

Figures

Fig. 1
Fig. 1
Histological appearance of CK-160 cervical carcinoma xenografts. Adjacent tumor sections stained with HE or immunostained for pimonidazole, HIF-1α, or CAIX shown at low (a) and high (b) magnification. The high magnification images refer to the squared box in (a)
Fig. 2
Fig. 2
Histological appearance of TS-415 cervical carcinoma xenografts. Adjacent tumor sections stained with HE or immunostained for pimonidazole, HIF-1α, or CAIX shown at low (a) and high (b) magnification. The high magnification images refer to the squared box in (a)
Fig. 3
Fig. 3
Pimonidazole and endogenous hypoxia markers. Tissue area fraction staining positive for pimonidazole, HIF-1α, or CAIX in CK-160 (a) and TS-415 (b) cervical carcinoma xenografts. Points, individual tumors; horizontal lines, mean values
Fig. 4
Fig. 4
Endogenous hypoxia markers and tumor hypoxia. PFHIF-1αversus HFPim, PFCAIXversus HFPim, and PFHIF-1αversus PFCAIX for CK-160 (a) and TS-415 (b) cervical carcinoma xenografts. Points, individual tumors
Fig. 5
Fig. 5
Endogenous hypoxia markers and tumor metastasis. HFPim, PFHIF-1α, or PFCAIX in metastatic and nonmetastatic CK-160 (a) and TS-415 (b) cervical carcinoma xenografts. Points, individual tumors; horizontal lines, mean values

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