Lactylation at K197 site of peroxiredoxin 1 promotes proliferation and migration of glioblastoma cells
10.3969/j.issn.1000-4718.2025.02.002
- VernacularTitle:过氧化还原蛋白1的K197位点乳酸化修饰促进胶质母细胞瘤细胞的增殖和迁移
- Author:
Guoliang DUAN
1
;
Qingliang HAN
;
Shibing FAN
Author Information
1. 重庆大学医学院,重庆 400000;重庆大学附属三峡医院神经外科,重庆 404000
- Publication Type:Journal Article
- Keywords:
glioblastoma;
peroxiredoxin 1;
lactylation;
point mutation;
cell proliferation;
cell migration
- From:
Chinese Journal of Pathophysiology
2025;41(2):219-229
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the effects of lactylation at the K197 site of peroxiredoxin 1(PRDX1)on the proliferation and migration of glioblastoma cells.METHODS:(1)Immunofluorescence and lactylation pan-antibody techniques were adopted to compare the differences in PRDX1 lactylation modification level between glioblastoma tissues and adjacent normal tissues.High-throughput mass spectrometry and modificomics analysis were utilized to select PRDX1 protein and its K197 site as the focus.(2)Cell experiments were conducted using lactate(5,10 and 15 mmol/L),glu-cose(5,10 and 25 mmol/L)and glycolysis inhibitor 2-deoxy-D-glucose(2-DG;1,5,10 and 15 mmol/L)to treat human glioblastoma U87MG and LN229 cells.Cell proliferation was detected by EdU proliferation staining,and PRDX1 expres-sion was detected in U87MG,LN229 and glial cells via immunoprecipitation and Western blot.The PRDX1 expression and lactylation levels were further examined in 10 mmol/L lactic acid-treated and untreated cells using immunoprecipita-tion and Western blot.(3)The U87MG and LN229 cells were transfected with constructed lactate dehydrogenase A(LDHA)siRNA(si-LDHA)plasmids and negative control(si-Con)plasmids,and the lactylation level of PRDX1 was as-sessed by immunoprecipitation and Western blot.(4)Similarly,the U87MG and LN229 cells were transfected with PRDX1 shRNA(sh-PRDX1)plasmids and negative control(sh-Con)plasmids,and PRDX1 expression was determined by Western blot.(5)The PRDX1 K197R mutant and PRDX1 wild-type(WT)plasmids were constructed and transfected into U87MG and LN229 cells.The PRDX1 expression and lactylation levels were determined by immunoprecipitation and Western blot.The CCK-8 and EdU assays were used to measure cell viability and proliferation,and Transwell assay was performed to assess the migration of U87MG and LN229 cells transfected with PRDX1 K197R mutant and PRDX1 WT plasmids.(6)A tumor formation model in nude mice was established.The LN229 cells with or without PRDX1 K197R mutation were used in the tumor formation experiment with 6 nude mice per group.After 18 d,the nude mice were eutha-nized,and tumor tissues were harvested.Histological changes were observed by HE staining,the lactylation modification leve was detected by immunofluorescence,and immunohistochemistry method was adopted for checking Ki67,a prolifera-tion marker,in tumor tissues.RESULTS:The PRDX1 level in glioblastoma tissues was significantly higher than that in adjacent tissues(P＜0.05).In cell experiments,the addition of lactate and glucose significantly promoted the proliferation and migration of glioblastoma cells and increased the lactylation level of PRDX1(P＜0.05).In contrast,the glycolysis in-hibitor 2-DG inhibited these effects.The si-LDHA transfection experiment showed that knockdown of LDHA reduced the lactylation level of PRDX1(P＜0.05).Importantly,the K197R point mutation in PRDX1 significantly decreased the lacty-lation level of PRDX1 and inhibited the proliferation and migration of glioblastoma cells(P＜0.05).Nude mouse tumori-genesis experiments further confirmed that tumor growth in PRDX1 K197R group was significantly reduced,and the Ki67 proliferation index and lactylation level were decreased(P＜0.05).CONCLUSION:Lactoylation at the K197 site of PRDX1 promotes the proliferation and migration of glioblastoma cells.
