# Hypoxia Triggers ALYREF‐Mediated m5C Methylation of KIF20A to Activate KIF20A/BUB1 for Generating Ferroptosis Resistance in Cervical Cancer Cells

## TL;DR

This study shows how hypoxia causes cervical cancer cells to resist cell death through a pathway involving KIF20A and BUB1, offering new insights into cancer treatment resistance.

## Contribution

The study identifies a novel mechanism involving ALYREF-mediated m5C methylation of KIF20A in hypoxia-induced ferroptosis resistance in cervical cancer.

## Key findings

- Hypoxia increases ferroptosis resistance in cervical cancer cells.
- ALYREF stabilizes KIF20A mRNA via m5C modification, promoting resistance.
- KIF20A activates BUB1, and BUB1 overexpression can partially reverse resistance.

## Abstract

Ferroptosis resistance is a key player in cervical cancer (CC) development. Hypoxia is a negative factor affecting CC treatment by inducing ferroptosis resistance. Our study aimed to investigate the detailed mechanisms of hypoxia‐induced ferroptosis resistance in CC cells. The mRNA and protein levels were assessed using RT‐qPCR and western blot. Immunofluorescence staining was used to analyze the co‐localization of Budding uninhibited by benzimidazole 1 (BUB1) and Kinesin family member 20A (KIF20A) in CC cells. Fe2+, MDA, and GSH levels were measured by commercial kits. Cell viability was determined by CCK‐8. The molecular interactions were analyzed by RIP or Co‐IP. MeRIP was adopted to measure the m5C level of KIF20A. We found that hypoxia facilitated ferroptosis resistance in CC cells. Hypoxia led to the upregulation of KIF20A, and KIF20A knockdown weakened hypoxia‐mediated ferroptosis resistance in CC cells. Mechanistically, Aly/REF export factor (ALYREF) stabilized KIF20A mRNA stability via m5C modification. In addition, KIF20A upregulated BUB1 in CC cells by directly interacting with BUB1. Rescue experiments indicated that BUB1 overexpression partially reversed the inhibitory effect of KIF20A knockdown on hypoxia‐mediated ferroptosis resistance in CC cells. In conclusion, hypoxia triggers ALYREF‐mediated m5C methylation of KIF20A to activate the KIF20A/BUB1 axis, thereby inducing ferroptosis resistance in CC cells.