STM2457通过降低m6A修饰水平调控铁死亡相关分子并抑制肝癌细胞恶性生物学行为OA

Objective To investigate the effects of STM2457 on N6-methyladenosine(m6A)modification level and ferroptosis-related molecules in liver cancer cells,as well as its role in the malignant biological behaviors of liver cancer cells.Methods Human liver cancer cells Huh-7 and mouse hepatoma cells Hepa1-6 were treated with different gradient concentrations of STM2457 to calculate the half maximal inhibitory concentration(IC50)in Huh-7 and Hepa1-6 cells.The STM2457 group(treated with STM2457 at the IC50 concentration)and the Control group(treated with DMSO)were established.The CCK-8 and colony formation assays were used to evaluate the effects of STM2457 on proliferation and clonogenicity of the two liver cancer cell lines.Transwell assay was performed to observe the effects of STM2457 on cell migration and invasion.Flow cytometry was used to detect the apoptosis rate.m6A dot blot assay was used to detect the effect of STM2457 on global m6A modification levels.Western blot was performed to detect the protein expression levels of ferroptosis-related molecules SLC7A11,GPX4,ACSL4,and METTL3.Results The IC50 of STM2457 was 139.8 μmol/L for Huh-7 cells and 95.79 μmol/L for Hepa1-6 cells.STM2457 significantly inhibited proliferation,colony formation,migration,and invasion of Huh-7 and Hepa1-6 cells,and promoted apoptosis(all P<0.05).After STM2457 treatment,global m6A levels decreased in both cell lines(both P<0.0001),while METTL3 protein expression did not change(all P>0.05).The expression levels of the core ferroptosis suppressor proteins SLC7A11 and GPX4 decreased,whereas the expression level of the key ferroptosis promoter protein ACSL4 increased(all P<0.01).Conclusions STM2457 may significantly reduce global m6A modification levels in liver cancer cells Huh-7 and Hepa1-6 without decreasing METTL3 protein expression,and alter the expression of ferroptosis-related molecules,thereby inhibiting the proliferation,migration,invasion,and clonogenicity of liver cancer cells in vitro.