Abstract P5-03-10: HIF-1alpha knockout radiosensitizes select Inflammatory Breast Cancer cells through reduction of stem-like cancer cells

Abstract

Abstract Independent of the tumor stroma, HIF-1α regulates tumor cell metabolism, cell proliferation and DNA repair pathways, which may produce conflicting responses to radiation, promoting either radioresistance or radiosensitization. Recently, HIF-1α was shown to increase tumor initiating cells (TICs) activity in normoxic tumor cells derived from the MMTV-PyMT trasngenic mouse model of luminal-like breast cancer. While TICs have been reported to be relatively radio-resistant, the role of HIF-1α in mediating the radation response in breast cancer TICs remains unknown. Herein, we examined radiosenstitivity of TICs derived from either PyMT HIF-1α knock out (KO) tumor cells or human breast cancer cell lines transduced with a HIF-1α dominant negative (DN) construct. Consistent prior results that loss of HIF-1 activity reduces TIC frequency, down-regulation of HIF-1 activity through the HIF DN construct reduced mammosphere formation in SUM-159, SUM-149 and MCF-7 cells (by 5.0-, 2.0-, and 2.0-fold respectively). Moreover, β-catenin expression was down-regulated in SUM-149 and SUM-159 cells transfected with the HIF-1α DN construct. Moreover, SUM-149 cells expressing the HIF-1α DN construct exhibited delayed tumor growth in vivo (p = 0.05). Standard clonogenic assays demonstrated that PyMT HIF-1α KO cells and SUM-149 cells expressing the HIF-1α DN construct were more sensitive to radiation therapy (SF6 of PyMT, HIF-1α:Control, 0.016:0.087), but that the PyMT KO and SUM-149 mammospheres that persisted after radiation were completely radioresistant (SF6 of PyMT, HIF-1 α: Control, 0.48:0.44). In contrast, MCF-7 cells were not radiosensitized in either standard or mammosphere assays. Interestingly, in HIF-1α DN MCF-7 cells, molecular features of IBC were observed, such as the increased expression of E-Cadherin and loss of Wisp3. But, Notch1 protein expression was unchanged between HIF-1α DN MCF7 or SUM149 cells. Moreover, concurrent radiation in the presence of a gamma secretase inhibitor or with a p53-MDM2 inhibitor nutlin failed to radiosensitize HIF KO mammosphere clonogens. We conclude that downregulation of HIF-1 activity selectively radiosensitizes IBC clonogenic cells but fails to radiosensitize the residual mammospheres. These data suggest that the known HIF-1α mediated mechanisms that favor radiosensitivity, such as the promotion of glycolysis and proliferation under stress, may predominate in mammospheres, which ultimately leads to radioresistance in residual mammospheres after HIF inhibition. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-03-10.