HIF-1 alpha switches the functionality of TGF-beta signaling via changing the partners of smads to drive glucose metabolic reprogramming in non-small cell lung cancer

Background: Most cancer cells have fundamentally different metabolic characteristics, particularly much higher glycolysis rates than normal tissues, which support the increased demand for biosynthesis and promote tumor progression. We found that transforming growth factor (TGF)-beta plays a dual function in regulating glycolysis and cell proliferation in non-small cell lung cancer. Methods: We used the PET/MRI imaging system to observe the glucose metabolism of subcutaneous tumors in nude mice. Energy metabolism of non-small cell lung cancer cell lines detected by the Seahorse XFe96 cell outflow analyzer. Co-immunoprecipitation assays were used to detect the binding of Smads and HIF-1 alpha. Western blotting and qRT-PCR were used to detect the regulatory effects of TGF-beta and HIF-1 alpha on c-MYC, PKM1/2, and cell cycle-related genes. Results: We discovered that TGF-beta could inhibit glycolysis under normoxia while significantly promoting tumor cells' glycolysis under hypoxia in vitro and in vivo. The binding of hypoxia-inducible factor (HIF)-1 alpha to the MH2 domain of phosphorylated Smad3 switched TGF-beta function to glycolysis by changing Smad partners under hypoxia. The Smad-p107-E2F4/5 complex that initially inhibited c-Myc expression was transformed into a Smad-HIF-1 alpha complex that promoted the expression of c-Myc. The increased expression of c-Myc promoted alternative splicing of PKM to PKM2, resulting in the metabolic reprogramming of tumor cells. In addition, the TGF-beta/Smad signal lost its effect on cell cycle regulatory protein p15/p21. Furthermore, high expression of c-Myc inhibited p15/p21 and promoted the proliferation of tumor cells under hypoxia. Conclusions: Our results indicated that HIF-1 alpha functions as a critical factor in the dual role of TGF-beta in tumor cells, and may be used as a biomarker or therapeutic target for TGF-beta mediated cancer progression.