灯盏花乙素抑制人前列腺癌细胞系PC-3的增殖和迁移

Scutellarin inhibits proliferationand migration of human prostate cancer cell line PC-3

目的探讨灯盏花乙素(STR)对人前列腺癌细胞系(PC-3)增殖、迁移的影响及其作用机制。方法PC-3细胞分为STR低、中和高剂量组、colivelin(STAT3激活剂)组、STR高剂量+colivelin组、对照组。CCK-8法、集落形成实验检测细胞增殖;划痕实验检测细胞迁移;透射电镜观察细胞线粒体超微结构;比色法检测细胞内游离Fe 2+、丙二醛(MDA)含量及活性氧(ROS)水平;RT-qPCR检测溶质载体家族7成员11(SLC7A11)、增殖细胞核抗原(PCNA)、基质金属蛋白酶(MMP)-9 mRNA表达;Western blot检测p-STAT3、GPX4蛋白。结果与对照组对比,STR低、中和高剂量组细胞线粒体结构破坏明显,A 450值、集落形成率、划痕愈合率、PCNA、SLC7A11、MMP-9 mRNA表达及p-STAT3、GPX4蛋白降低(P<0.05),Fe 2+、MDA含量及ROS水平升高,且呈剂量依赖性(P<0.05);与对照组相比,colivelin组细胞中线粒体结构破坏有所改善,A 450值、集落形成率、划痕愈合率、PCNA、SLC7A11、MMP-9 mRNA表达及p-STAT3、GPX4蛋白升高,Fe 2+、MDA含量及ROS水平降低(P<0.05);与STR高剂量组相比,STR高剂量+colivelin组细胞中线粒体结构破坏有所减轻,A 450值、集落形成率、划痕愈合率、PCNA、SLC7A11、MMP-9 mRNA表达及p-STAT3、GPX4蛋白升高,Fe 2+、MDA含量及ROS水平降低(P<0.05)。结论STR降低和减弱前列腺癌细胞增殖和迁移能力,其机制可能与抑制STAT3/GPX4通路有关。

Objective To investigate the effect of scutellarin(STR)on the proliferation and migration of human prostate cancer cell line(PC-3)and its underlying mechanism.Methods PC-3 cells were divided into low-dose STR group,medium-dose STR group,high-dose STR group,colivelin(STAT3 activator)group,high-dose STR+colivelin group and control group.CCK-8 assay and colony formation experiments were applied to detect cell proliferation;Scratch experiment was applied to detect cell migration;Transmission electron microscopy was applied to observe the mitochondrial ultrastructure of PC-3 cells.The intracellular free Fe 2+,malondialdehyde(MDA)content and reactive oxygen species(ROS)levels were detected by colorimetric method;RT-qPCR was applied to detect the mRNA expression of member 11 of solute vector family(SLC7A11),proliferating cell nuclear antigen(PCNA)and matrix metalloproteinase-9(MMP-9)in cells;Western blot was used to detected p-STAT3 and GPX4 proteins in cells.Results Compared to control group,the mitochondrial structure of PC-3 cells in the low-dose STR group,medium-dose STR group and high-dose STR group was significantly disrupted.The A 450 value,colony formation rate,scratch healing rate,PCNA,SLC7A11,MMP-9 mRNA expression,and p-STAT3,GPX4 protein all reduced.While Fe 2+,MDA content,and that ROS level increased with dose-dependent way(P<0.05).Compared with control group,the destruction of mitochondrial structure in cells from colivelin group improved;The A 450 value,colony formation rate,scratch healing rate,PCNA,SLC7A11,MMP-9 mRNA expression and p-STAT3,GPX4 protein all increased,while Fe 2+MDA content,and ROS level decreased(P<0.05).Compared with high-dose STR group,the damage of mitochondrial structure in PC-3 cells in the high-dose STR+colivelin group was reduced.The A 450 value,colony formation rate,scratch healing rate,PCNA,SLC7A11,MMP-9 mRNA expression,and p-STAT3,GPX4 protein increased,while Fe 2+,MDA content and ROS level decreased(P<0.05).Conclusions The mechanism by of STR reducing proliferation and migration of prostate cancer cells is potentially related to the inhibition of STAT3/GPX4 pathway.