Effect of IGF2BP2 inhibitor CWI1-2 on glioma migration, invasion and TMZ resistance
10.3760/cma.j.cn115354-20240924-00592
- VernacularTitle:IGF2BP2抑制剂CWI1-2对胶质瘤细胞迁移、侵袭能力及替莫唑胺耐药性的影响
- Author:
Yilamu YIMURAN
1
;
Zhennan TAO
;
Yi SUN
;
Chuhua HANG
Author Information
1. 南京大学医学院，南京 210008
- Publication Type:Journal Article
- Keywords:
Glioma;
IGF2BP2;
CWI1-2;
Migration;
Invasion;
Temozolomide
- From:
Chinese Journal of Neuromedicine
2025;24(2):131-140
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect of insulin-like growth factor 2 ( IGF2) mRNA binding protein 2 (IGF2BP2) inhibitor CWI1-2 on migration and invasion of glioma cells SHG-140 and LN229 as well as their resistance to temozolomide (TMZ). Methods:(1) SHG-140 and LN229 glioma cell lines were cultured in vitro; cell counting kit-8 (CCK-8) assay was used to detect the effects of 0.1, 0.2, 0.5, 1.0, 2, and 5 μmol/L CWI1-2 on survival rate of the two cell lines at 24 and 48 hours after treatment, and the best concentration and treatment time of CWI1-2 were screened. SHG-140 and LN229 cells were categorized into control group, 0.5 μmol/L CWI1-2 group, and 1.0 μmol/L CWI1-2 group, respectively, and equal amount of medium, 0.5 μmol/L CWI1-2 and 1.0 μmol/L CWI1-2 were added into the cells for 24 hours; Western blotting and immunofluorescent staining were used to detect the IGF2BP2 protein expression; cell scratch assay and Transwell assay were, respectively, used to detect the cell migration and invasion. (2) CCK-8 assay was employed to detect the effects of 100, 200, 400, 600, 800, and 1000 μmol/L TMZ on survival rate of SHG-140 and LN229 cells at 24 and 48 hours after treatment, and the best concentration and treatment time of TMZ were screened. SHG-140 and LN229 cells were categorized into control group, CWI1-2 group, TMZ group, and TMZ+CWI1-2 group, respectively, and equal amounts of culture medium, 0.5 μmol/L CWI1-2, 200 μmol/L TMZ, and 0.5 μmol/L CWI1-2+200 μmol/L TMZ were added into the 4 groups for 24 hours, respectively; colony formation assay was used to detect the colony formation rate and CCK-8 assay was utilized to determine the proliferation rate. Results:(1) CCK-8 results indicated that, compared with the 0.1 and 0.2 μmol/L CWI1-2 groups, SHG-140 and LN229 cells in the 0.5, 1.0, 2, and 5 μmol/L CWI1-2 groups had significantly lower survival rate at 24 and 48 hours after treatment ( P<0.05); furthermore, the survival rate in the SHG-140 and LN229 cells of the 0.5, 1.0, 2, and 5 μmol/L CWI1-2 groups decreased successively, with statistical differences ( P<0.05); therefore, 0.5 μmol/L and 1 μmol/L CWI1-2 for 24 hours with less effect on survival rate were selected for subsequent experiments. Western blotting results showed that, compared with the control group (1.27±0.05), the SHG-140 cells in 0.5 and 1.0 μmol/L CWI1-2 groups had significantly lower IGF2BP2 protein expression (0.91±0.09, 0.79±0.10; P<0.05); compared with the control group and 0.5 μmol/L CWI1-2 group (0.82±0.09, 0.82±0.04), the LN229 cells in 1.0 μmol/L CWI1-2 group had significantly lower IGF2BP2 protein expression (0.33±0.02, P<0.05). Immunofluorescent staining results showed that, compared with the control group, the 0.5 and 1.0 μmol/L CWI1-2 groups had obviously decreased IGF2BP2 protein immunofluorescent intensity. Cell scratch assay results showed that, compared with the control group, the SHG-140 cells in 0.5 and 1.0 μmol/L CWI1-2 groups had significantly lower migration rate ( P<0.05); the migration rate in LN229 cells of the control group, 0.5 μmol/L CWI1-2 group and 1.0 μmol/L CWI1-2 group decreased successively, with statistical differences ( P<0.05). Transwell assay results showed that the number of transmembrane SHG-140 and LN229 cells in the control group, 0.5 μmol/L CWI1-2 group and 1.0 μmol/L CWI1-2 group decreased successively, with significant differences ( P<0.05). (2) CCK-8 assay results showed that compared with the 100 and 200 μmol/L TMZ groups, SHG-140 and LN229 cells in the 400, 600, 800 and 1000 μmol/L TMZ groups had statistically lower survival rate at 24 and 48 hours after treatment ( P<0.05); therefore, concentration with less effect on cell survival rate (200 μmol/L TMZ) was selected for subsequent experiments. The cell colony formation rate of SHG-140 and LN229 cells in the control group, CWI1-2 group, TMZ group and TMZ+CWI1-2 group decreased successively, with significant differences ( P<0.05). Cell proliferation rate of LN229 cells in the control group, CWI1-2 group, TMZ group and TMZ+CWI1-2 group decreased successively, with significant differences ( P<0.05). Conclusion:CWI1-2 can inhibit the proliferation and invasion of glioma cells, and can also increase the killing effect of TMZ on glioma cells.
