Six-week-old male C57BL/6J mice were purchased from the Medical Laboratory Animal Center of Guangdong Province (Foshan, China). Forty-five C57BL/6L mice were randomized into 4 groups after 1 week of adaptive feeding: (1) Control group (n = 9); (2) AOM/DSS group (n = 12); (3) AOM/DSS + NaB (orally) group (n = 12); 4) AOM/DSS + NaB (intraperitoneal injection) group (n = 12). The Control group received an intraperitoneal injection of saline solution beginning on day 1, and received sterile drinking water throughout the study. Other three groups received an intraperitoneal injection of 10 mg/kg AOM (Sigma Aldrich) beginning on day 1, and received drinking water containing 2.5% DSS at the second and eighth weeks (2% DSS in the fifth week). Besides, 0.1 M NaB (Sigma Aldrich) was given in drinking water during the whole experiment process in AOM/DSS + NaB (p.o.) group, while AOM/DSS + NaB (i.p.) group was injected intraperitoneally (IP) with 1 g/kg NaB per day.

ALI induction was performed in 6-8-week-old age-matched C57BL/6J male mice (n = 10-12 per group) by intraperitoneal injection of 3 mL/kg CCl4 in coconut oil. Control and negative control mice were injected with PBS and coconut oil, respectively. At 6 h after injection of CCl4, mice were divided into three groups: CCl4 group, injected with 100 uL PBS (supplemented with 2% mouse serum) through a tail vein; CCl4 + MSC group, injected with 5 x 10⁵ MSCs suspended in 100 uL PBS (supplemented with 2% mouse serum) through a tail vein; CCl4 + Fer-1 group, intraperitoneally injected with ferrostatin-1 (Fer-1, a ferroptosis inhibitor, 2.5 umol/kg body weight). Erastin, intraperitoneal injection of erastin (a ferroptosis inducer, 30 mg/kg body weight) twice every other day, and then the mice were divided into two groups (n = 10-12 per group): Erastin group, injected with 100 uL PBS (supplemented with 2% mouse serum) through a tail vein; Erastin + MSC group, injected with 5 x 10⁵ MSCs suspended in 100 uL PBS (supplemented with 2% mouse serum) through the tail vein.

5.0 x 10⁶ cells were mixed with Matrigel (BD Biosciences) at 1:1 ratio (v/v) and injected subcutaneously into seven-week old nude mice (NU/NU; Charles River). Mice were fed with regular chow. After nine weeks, the mice were killed and the tumors were weighed and recorded.

In this study, 4-week-old specific pathogen free male nude mice were selected and assigned into 4 groups, each of which was respectively injected with U251 cells and U87 cells transfected with NC, hsa-miR-27a-3p angomir, si-TMEM161B-AS1, si-TMEM161B-AS1 + hsa-miR-27a-3p angomir. Each nude mouse was subcutaneously injected with 4 x 10⁵ cells in the right flank area for subcutaneous implantation.