薯蓣皂苷增强DNA损伤促进口腔鳞癌细胞凋亡

Dioscin Enhances DNA Damage to Promote Apoptosis in Oral Squamous Cell Carcinoma Cells

目的:研究薯蓣皂苷对人口腔鳞癌细胞SCC4、SCC25 DNA损伤和凋亡的影响及其可能的机制。方法:SCC4、SCC25细胞用不同浓度薯蓣皂苷(0.0、0.1、0.3、1.0、3.0、10.0和30.0μmol/L)处理48 h,通过CCK-8实验检测细胞活力;不同浓度(0.0、1.0、2.0、4.0μmol/L)薯蓣皂苷处理SCC4细胞48 h,采用免疫荧光技术检测γH2AX在SCC4细胞核内的表达情况;通过Western blotting检测DNA损伤修复蛋白的表达情况;采用流式细胞术检测细胞凋亡率,最后通过Western blotting检测细胞凋亡相关蛋白的表达变化。结果:薯蓣皂苷浓度依赖性地抑制了SCC4和SCC25细胞的活性(P<0.05);与薯蓣皂苷0.0μmol/L组比较,各薯蓣皂苷处理组SCC4细胞核中γH2AX的荧光表达强度显著上调(P<0.05);与薯蓣皂苷0.0μmol/L组比较,各薯蓣皂苷处理组SCC4、SCC25细胞DNA损伤信号蛋白P-ATM、P-CHK1、P-CHK2、γH2AX表达上升;与薯蓣皂苷0.0μmol/L组比较,各薯蓣皂苷处理组SCC4、SCC25细胞Cleaved-PARP、Cleaved-Caspase-3蛋白表达均升高。结论:薯蓣皂苷能抑制口腔鳞癌细胞的活性,并促进细胞的凋亡,其作用机制可能和其促进细胞的DNA损伤有关。

Objective:To investigate the effects of Dioscin on DNA damage and apoptosis in SCC4 and SCC25 human OSCC cells and explore potential underlying mechanisms.Methods:SCC4 and SCC25 cells were treated with various concentrations of Dioscin(0.0,0.1,0.3,1.0,3.0,10.0 and 30.0μmol/L)for 48 hours,and cell viability was assessed using the CCK-8 assay.Immunofluorescence was used to evaluateγH2AX expression in SCC4 cell nuclei after treatment with different Dioscin concentrations(0.0,1.0,2.0,4.0μmol/L)for 48 hours.The expression of DNA damage repair proteins was examined by Western blotting.Flow cytometry was employed to measure apoptosis rates,and changes in the expression of apoptosis-related proteins were detected using Western blotting.Results:Dioscin exhibited a concentration-dependent inhibition of SCC4 and SCC25 cell activity(P<0.05).It significantly increased the fluorescence intensity ofγH2AX in SCC4 cell nuclei compared to the untreated group(P<0.05)and activated the expression of DNA damage signal proteins,including P-ATM,P-CHK1,P-CHK2,andγH2AX.Dioscin also dose-dependently facilitated the activation of apoptosis-regulating proteins Cleaved-PARP and Cleaved-Caspase-3.Conclusion:Dioscin can suppress the activity of oral squamous cell carcinoma cells and promote apoptosis,and its mechanism may be linked to its induction of DNA damage in these cells.