Hydrogen gas promotes apoptosis of lung adenocarcinoma A549 cells through X-linked inhibitor of apoptosis and baculoviral inhibitor of apoptosis protein repeat-containing 3.

Abstract

Lung cancer is currently the cancer with the highest incidence and death toll worldwide. Hydrogen gas has been found to affect a variety of diseases; however, the effect of hydrogen gas on patients with lung cancer has not been reported. Therefore, we determined the effect of hydrogen gas on apoptosis of lung adenocarcinoma in vivo and in vitro. A549 cells in the logarithmic phase were treated with 20%, 40%, or 60% hydrogen gas. Cell apoptosis was evaluated by flow cytometry. The A549 cell suspension was inoculated into 15 nude mice. The mice were randomly divided into control, hydrogenation (inhalation of 60% hydrogen gas), and cisplatin groups (intraperitoneal injection of cisplatin [4 mg/kg]). After 3 weeks, the tumor tissue was removed and measured. We identified differentially expressed genes by transcriptional profiling. The levels of X-linked inhibitor of apoptosis (XIAP), baculoviral inhibitor of apoptosis protein repeat-containing 3 (BIRC3), and BCL2-associated X and apoptosis regulator (BAX) protein expression were detected by Western blotting and immunohistochemistry. Compared with the control group, the apoptosis rates in the 20%, 40%, and 60% hydrogen gas groups were significantly increased (P < 0.01). The levels of XIAP and BIRC3 protein expression were clearly decreased in the hydrogen gas group compared to the control group. Moreover, cisplatin and hydrogen gas reduced the tumor volume in nude mice (P < 0.01). Transcriptome sequencing showed that XIAP, BIRC2, BIRC3, BAX, PIK3CD, and ATM were related to apoptosis. Hydrogen gas further decreased the levels of XIAP and BIRC3 expression than in nude mice (P < 0.01). Hydrogen gas promoted apoptosis of A549 cells by reducing the expression of XIAP and BIRC3 protein.