RNA binding protein LIN28B promotes chemosensitivity of colon cancer by regulating the synthesis and activity of glutathione
10.3760/cma.j.cn113855-20240202-00100
- VernacularTitle:RNA结合蛋白LIN28B通过调控谷胱甘肽的合成和活性促进结肠癌细胞的化疗敏感性
- Author:
Ning NING
1
;
Yeqing SONG
;
Yichao YAN
;
Lin CHEN
;
Yankai ZHANG
Author Information
1. 北京大学国际医院胃肠外科，北京 102206
- Publication Type:Journal Article
- Keywords:
Colonic neoplasms;
Glutathione;
Drug resistance,neoplasm;
Antineoplastic combined chemotherapy protocols
- From:
Chinese Journal of General Surgery
2025;40(8):643-649
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the influence of LIN28B on chemosensitivity of colon cancer by regulating GSH.Methods:Functional enrichment analysis of LIN28B target genes was performed using database. The primary tumor tissues of colon cancer patients who received neoadjuvant chemotherapy at Department of Gastroenterology, Peking University International Hospital from Nov 2017 to May 2020 were collected, and their LIN28B levels were detected by immunohistochemistry. According to the tumor regression grade, they were divided into chemotherapy sensitive group and chemotherapy resistant group, and the difference of LIN28B expression between the two groups was compared. LIN28B overexpression and knockdown colon cancer cell lines were constructed, and the effect of LIN28B on the chemosensitivity of colon cancer cells was detected by MTT assay. Double luciferase reporting experiment and Western blot were used to detect the direct binding and regulation of LIN28B to mRNA of four GSH related enzymes. At the same time, the regulation of LIN28B on total GSH and reduced GSH was tested. Finally, by detecting the level of reactive oxygen species (ROS) γ-H2AX and Comet assay to analyze the potential impact of LIN28B on genomic instability.Results:GSH-related enzymes were highly enriched in LIN28B target genes. The expression of LIN28B was heterogeneous in colon cancer patients. Compared with the low expression group, the average survival time of patients with high expression of LIN28B was significantly increased [(50.2±2.9 )months vs. (31.1±4.0 )months, P=0.001], and the proportion of tumor regression grade 0-1 was significantly different (48.0% vs. 16.0%, P=0.032). The expression level of LIN28B in chemotherapy sensitive group was significantly higher than that in drug resistant group ( P<0.01). LIN28B overexpression significantly increased the chemosensitivity of HCT116 cells to 5-fluorouracil (5-Fu) and oxaliplatin (L-OPH). The synthesis and activity of GSH were further inhibited (all P<0.01). At the same time, the ROS level of LIN28B overexpression cells was significantly increased after treatment with L-OPH. The level of γ-H2AX was significantly increased, and the content of comet tail DNA was also significantly increased ( P<0.01). Conclusion:LIN28B may increase the chemosensitivity of colon cancer cells by directly inhibiting the expression of GSH related enzymes, resulting in the decrease of GSH synthesis and activity, the increase of ROS level and genomic instability.
