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Hypoxia inducible factor-1α/B-cell lymphoma 2 signaling impacts radiosensitivity of H1299 non-small cell lung cancer cells in a normoxic environment

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Abstract

Hypoxia inducible factor-1α (HIF-1α) is a critical transcriptional factor for the response of cells to hypoxic microenvironment and its expression induces resistance of hypoxic non-small-cell lung cancer (NSCLC) cells to radiotherapy. This study investigated how the activation of HIF-1α/B-cell lymphoma 2 (BCL-2) signaling under normoxic conditions impacted radiosensitivity of NSCLC cells. The recombinant pcDNA3.0-EGFP plasmids with wild-type or mutant HIF-1α complementary DNA (cDNA) were transfected into H1299 cells, an NSCLC cell line, establishing two H1299 sublines with high expression of HIF-1α. Compared with the levels of HIF-1α and BCL-2 proteins in non-transfected cells, increased levels of both proteins were found in transfected cells. Moreover, the expression of HIF-1α in non-transfected cells induced by chloride cobalt (CoCl2), a commonly used mimetic hypoxia reagent, was concomitant with the enhancement of BCL-2 expression. Conversely, reduction of HIF-1α expression by an inhibitor decreased the levels of BCL-2 proteins. The results revealed that the stabilization and expression of HIF-1α promoted the accumulation of BCL-2 proteins in H1299 cells. Subsequent experiments showed that intracellular HIF-1α/BCL-2 signaling was triggered in a normoxic environment after H1299 cells were exposed to irradiation, causing an elevated radioresistance. In contrast, blockage of HIF-1α/BCL-2 signaling leads to an elevated radiosensitivity. Proliferation of cells assay showed that, under normoxic conditions, population doubling times (PDTs) of irradiated cells were prolonged by suppression of HIF-1α/BCL-2 signaling. It is, therefore, indicated that HIF-1α/BCL-2 signaling activated by ionizing radiation reduces the radiosensitivity of H1299 cells independent of the hypoxic environment.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant Nos. U1432124 and 81673099). We appreciate the assistance by Ms. Micha Davila (Centre for Molecular Immunology and Infectious Disease, the Pennsylvania State University) for improvement of the English of this paper.

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  1. Gang Wang and Liang Xiao have contributed equally to this work.

Authors and Affiliations

  1. Teaching and Research Section of Nuclear Medicine, Anhui Medical University, 81 Meishan Road, Hefei, 230032, Anhui, People’s Republic of China

    Gang Wang, Liang Xiao, Fen Wang, Jing Yang, Li Yang, Ye Zhao & Wensen Jin

  2. Department of Radiation Oncology, First Affiliated Hospital, Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, People’s Republic of China

    Liang Xiao

  3. Department of Radiation Oncology, Anhui Provincial Cancer Hospital, 107 Huanhu East Road, Hefei, 230031, Anhui, People’s Republic of China

    Jing Yang

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  1. Gang Wang
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Wang, G., Xiao, L., Wang, F. et al. Hypoxia inducible factor-1α/B-cell lymphoma 2 signaling impacts radiosensitivity of H1299 non-small cell lung cancer cells in a normoxic environment. Radiat Environ Biophys 58, 439–448 (2019). https://doi.org/10.1007/s00411-019-00802-4

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