Helium low temperature plasma induced HepG2 cells autophagy through ROS-mediated PI3K/AKT/mTOR/P70s6k signaling pathway

Low temperature plasma (LTP) has emerged as a new research hotspot of promising therapy to fight against cancer. Autophagy is a multifunctional process that digests and recycles cellular contents within lysosomes to maintain homeostasis, which is the underlying mechanism of cancer cell death and anti-cancer treatment. This study aimed at investigating whether autophagy of HepG2 cells could be induced by LTP and revealing the related molecular mechanism. For this purpose, the atmospheric pressure plasma jet (APPJ) in helium was utilized to generate plasma-actived medium (PAM) to treat HepG2 cells in vitro. Catalase (CAT), superoxide dismutase (SOD) and 3-methyladenine (3-MA) were respectively added into the PAM as the intervention group. The cell viability, formation of autophagosomes, intracellular reactive oxygen species (ROS) level, expression levels of autophagy-associated proteins and key proteins involved in PI3K/AKT/mTOR pathways were detected. The results showed that LTP inhibited cell viability in a dose- and time-dependent manner. Autophagy was induced through the formation of autophagosomes, conversion of LC3-II/LC3-I, increased expression of beclin 1 and degradation of p62. The mechanism was deduced that LTP enhanced the intracellular ROS level and decreased the phosphorylation level of key proteins in PI3K/AKT/mTOR/p70S6K pathway. However, these effects were blocked by the autophagy inhibitor 3-MA and ROS scavengers (CAT and SOD). The therapeutic efficacy of LTP against HepG2 cells may involve autophagy via suppressing PI3K/AKT/mTOR signaling pathway by LTP-mediated ROS.