Role of Mfn2-mediated mitochondrial homeostasis in hypoxia-reoxygenation injury in human colorectal adenocarcinoma cells
10.3760/cma.j.cn131073-20240526-00509
- VernacularTitle:Mfn2介导的线粒体稳态在人结直肠腺癌细胞缺氧复氧损伤中的作用
- Author:
Ziyi WENG
1
;
Rong CHEN
1
;
Qingtao MENG
1
Author Information
1. 武汉大学人民医院麻醉科，武汉 430060
- Publication Type:Journal Article
- Keywords:
Mitochondrial proteins;
Homeostasis;
Reperfusion injury;
Intestines
- From:
Chinese Journal of Anesthesiology
2025;45(5):569-573
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate the role of mitochondrial fusion protein 2 (Mfn2)-mediated mitochondrial homeostasis in hypoxia-reoxygenation injury in human colorectal adenocarcinoma cells.Methods:Human colorectal adenocarcinoma cell line Caco-2 was cultured in vitro and divided into 4 groups ( n=30 each) using a random number table method: control group (C group), hypoxia-reoxygenation group (HR group), transfection with negative control virus plus hypoxia-reoxygenation group (Vector+ HR group) and transfection with Mfn2 plus hypoxia-reoxygenation group (Mfn2+ HR group). Mfn2 was packaged by lentivirus and transfected into Caco-2 cells, and the hypoxia-reoxygenation model was established through exposing cells to hypoxia for 12 h followed by 2 h reoxygenation. The cell viability and levels of lactate dehydrogenase (LDH) were measured. The expression of Mfn2, Occludin, Claudin-1, E-cadherin and microtubule-associated protein 1 light chain 3 Ⅰ (LC3Ⅰ), LC3Ⅱ, P62, translocase of outer mitochondrial membrane (TOM20) and cytochrome c oxidase Ⅳ (COX Ⅳ) was detected by Western blot. The levels of ATP content and ROS were determined. Results:Compared with C group, the cell viability and ATP content were significantly decreased, the LDH activity and ROS level were increased, the expression of Mfn2, Occludin, Claudin-1 and E-cadherin was down-regulated, the expression of LC3Ⅱ was up-regulated, and the expression of P62, TOM20 and COX Ⅳ was down-regulated in HR group ( P<0.05). Compared with HR group, the cell viability and ATP content were significantly increased, the LDH activity and ROS level were decreased, the expression of Mfn2, Occludin, Claudin-1 and E-cadherin was up-regulated, the expression of LC3Ⅱ was down-regulated, and the expression of P62, TOM20 and COX Ⅳ was up-regulated in Mfn2+ HR group ( P<0.05), and no significant change was found in the aforementioned parameters in Vector+ HR group ( P>0.05). Conclusions:Mfn2 may alleviate hypoxia-reoxygenation injury in human colorectal adenocarcinoma cells by maintaining mitochondrial homeostasis.
