丹参酚酸B调控ROS抑制大肠癌细胞HCT-116增殖并促进其凋亡

目的 验证丹参酚酸B(Salvianolic acid B,SalB)抑制人大肠癌细胞HCT-116增殖并促进其凋亡,进一步通过活性氧簇(Reactive oxygen species,ROS)阐述其可能机制。方法 体外培养大肠癌细胞HCF-116,分成HCT-116组、HCT-116+H₂O₂组、HCT-116+SalB组、HCT-116+SalB+N-乙酰-L-半胱氨酸(N-acetylcysteine,NAC)组。分组干预后,采用MTT法检测细胞存活率、平板克隆实验检测细胞增殖、流式细胞术检测ROS含量、细胞周期及细胞凋亡率。结果 SalB对HCT-116细胞具有抑制作用,且在一定浓度范围内呈正相关(P＜0.01);SalB、H₂O₂促进HCT-116细胞内ROS生成(P＜0.01),ROS清除剂NAC预处理可清除由SalB产生的ROS(P＜0.01);SalB抑制HCT-116细胞增殖(P＜0.01)并促进其凋亡(P＜0.01),该作用可被NAC部分逆转(P＜0.05);SalB引起HCT-116细胞G0/G1周期阻滞(P＜0.01),NAC预处理完全逆转SalB导致的周期阻滞(P＜0.05)。结论 SalB可通过增加HCT-116细胞内ROS水平引起细胞周期阻滞,从而抑制细胞增殖,并促进其凋亡。

Salvianolic acid B regulates ROS to inhibit the proliferation and promote apoptosis in colorectal cancer HCT-116 cells

Objective The aims of this study were to verify that Salvianolic acid B(SalB)inhibited the proliferation of human colorectal cancer HCT-116 cells and promoted its apoptosis,and further elaborated its possible mechanism through reactive oxygen species(ROS). Methods The experiment was divided into four groups:HCT-116 group,HCT-116+H202 group,HCT-116+SalB group,and HCT-116+SalB+N-acetylcysteine(NAC)group.After group intervention,MTT assay was used to detect cell viability,plate colony formation assay to detect cell proliferation,and flow cytometry to detect ROS content,cell cycle and apoptotic rate. Results SalB had an inhibitory effect on HCT-116 cells,and it was a positively correlated within a certain concentration range(P＜0.01);SalB and H₂O₂ promoted the generation of ROS in HCT-116 cells(P＜0.01),and pretreatment with ROS scavenger NAC could eliminate ROS produced by SalB(P＜0.01);SalB inhibited the proliferation of HCT-116 cells(P＜0.01)and promoted apoptosis(P＜0.01).This effect could be partially reversed by NAC(P＜0.05);SalB caused cycle arrest at the G0/G1 phase in HCT-116 cells(P＜0.01),and NAC pretreatment completely reversed the cell cycle arrest induced by SalB(P＜0.05). Conclusion SalB can cause cell cycle arrest by increasing the level of ROS in HCT-116 cells,thereby inhibiting cell proliferation and promoting cell apoptosis.