TGF-β2 initiates autophagy via Smad and non-Smad pathway to promote glioma cells' invasion

Background: Glioblastoma multiforme (GBM) is characterized by lethal aggressiveness and patients with GBM are in urgent need for new therapeutic avenues to improve quality of life. Current studies on tumor invasion focused on roles of cytokines in tumor microenvironment and numerous evidence suggests that TGF-beta 2 is abundant in glioma microenvironment and vital for glioma invasion. Autopagy is also emerging as a critical factor in aggressive behaviors of cancer cells; however, the relationship between TGF-beta 2 and autophagy in glioma has been poorly understood. Methods: U251, T98 and U87 GBM cell lines as well as GBM cells from a primary human specimen were used in vitro and in vivo to evaluate the effect of TGF-beta 2 on autophagy. Western blot, qPCR, immunofluorescence and transmission-electron microscope were used to detect target molecular expression. Lentivirus and siRNA vehicle were introduced to establish cell lines, as well as mitotracker and seahorse experiment to study the metabolic process in glioma. Preclinical therapeutic efficacy was evaluated in orthotopic xenograft mouse models. Results: Here we demonstrated that TGF-beta 2 activated autophagy in human glioma cell lines and knockdown of Smad2 or inhibition of c-Jun NH2-terminal kinase, attenuated TGF-beta 2-induced autophagy. TGF-beta 2-induced autophagy is important for glioma invasion due to the alteration of epithelial-mesenchymal transition and metabolism conversion, particularly influencing mitochondria trafficking and membrane potential (Delta psi m). Autopaghy also initiated a feedback on TGF-beta 2 in glioma by keeping its autocrine loop and affecting Smad2/3/7 expression. A xenograft model provided additional confirmation on combination of TGF-beta inhibitor (Galunisertib) and autophagy inhibitor (CQ) to better turn off tumor growth. Conclusion: Our findings elucidated a potential mechanism of autophagy-associated glioma invasion that TGF-beta 2 could initiate autophagy via Smad and non-Smad pathway to promote glioma cells' invasion.