Effects of KAI gene expression on ferroptosis in pancreatic cancer cells.

Abstract

The specific role and mechanism of ferroptosis in the development of pancreatic cancer(PC) remain to be elucidated. The present study aimed to investigate the effects of the overexpression of the KAI1 gene on the ferroptosis of the human PC cell line MIA PaCa‑2. MIA PaCa‑2 cells infected with pCMV‑KAI1 and selected by G418 and KAI1 protein were analyzed by western blotting. The MIA PaCa‑2 cells with a stable expression of the KAI1 gene were termed MIA PaCa‑2‑KAI1. The proliferative capacities of MIA PaCa‑2 and MIA PaCa‑2‑KAI1 cells were detected using Cell Counting Kit‑8. The reactive oxygen species(ROS) in the cells were compared by flow cytometry. The expressions of ferroportin(FPN) and glutathione peroxidase4(GPX4) protein were analyzed by western blotting. The KAI1 stable expression cell line was confirmed and relabeled as MIA PaCa‑2‑KAI1. No significant differences in the proliferation of MIA PaCa‑2 and MIA PaCa‑2‑KAI1 were identified. Following treatment with a ferroptosis blocker, the increase in the proliferation of MIA PaCa‑2‑KAI1 (from 2.06±0.02 to 2.75±0.02) was more evident compared with MIA PaCa‑2 (from 2.94±0.02 to 2.95±0.02; P<0.05). The ROS in MIA PaCa‑2‑KAI1 was significantly higher compared with MIA PaCa‑2 (P<0.05). FPN and GPX4 protein demonstrated higher expression levels in MIA PaCa‑2‑KAI1 compared with MIA PaCa‑2. Moreover, KAI1 exerted an obvious promotion effect on FPN expression. This study identified that the high expression of the KAI1 gene promoted the occurrence of ferroptosis in PC cells through its extensive effect on FPN and GPX4. KAI1‑induced ferroptosis did not significantly inhibit the proliferation of PC cells.