小白菊内酯通过下调NOX4抑制结肠癌SW480细胞增殖和侵袭

目的:探讨小白菊内酯对人结肠癌细胞SW480增殖及侵袭的影响,并探讨其抗肿瘤的可能作用机制。方法:以不同浓度(5,10,15μmol·L^-1)小白菊内酯作用于体外培养的人结肠癌SW480细胞,以环磷酰胺(30μmol·L^-1)作为阳性对照。采用CCK8法以及平板克隆法检测细胞增殖,Transwell法检测细胞迁移及侵袭;免疫印迹法(Western blotting)检测NOX4蛋白以及增殖相关蛋白β-连环蛋白(β-catenin)、G1/S-特异性周期蛋白-D1(Cyclin D1)、侵袭相关蛋白金属基质蛋白酶2(MMP-2)、金属基质蛋白酶9(MMP-9)蛋白]表达情况。结果:随着小白菊内酯浓度的增加,对SW480细胞增殖抑制率明显升高,与空白组比较,差异有统计学意义(P<0.05);同一浓度作用下,随作用时间的延长,增殖抑制率显著升高(P<0.05),小白菊内酯10,15μmol·L^-1浓度组与阳性对照组比较差异有统计学意义(P<0.05);与空白对照组相比,小白菊内酯组克隆细胞形成数、侵袭、迁移细胞数、增殖相关蛋白β-catenin、Cyclin D1、侵袭迁移关蛋白MMP-2、MMP-9以及NOX4蛋白水平显著下降(P<0.05),且小白菊内酯各组间呈浓度依赖(P<0.05);与阳性对照组相比,小白菊内酯10,15μmol·L^-1浓度组以上各指标差异有统计学意义(P<0.05)。结论:小白菊内酯可能通过下调NOX4表达,抑制结肠癌细胞SW480细胞增殖、侵袭能力。

Inhibition of Parithenolide against the Proliferation and Invasion of Colon Cancer SW480 Cells by Down-regulating NOX4

Objective:To investigate the effects of parithenolide on the proliferation and invasion of human colon cancer cell line SW480,and explore its possible anti-tumor mechanism.Methods:Human colon cancer SW480 cells were cultured in vitro with different concentrations of parithenolide(5,10 and 15μmol·L^-1),and cyclophosphamide(30μmol·L^-1)was used as a positive control.CCK8 assay and plate cloning assay were used to detect the cell proliferation,the cell migration and invasion were detect by Transwell method,Western blot was used to detect the expressions of NOX4 protein and proliferation-related proteinsβ-catenin,Cyclin D1,invasion-related protein metalloproteinase-2(MMP-2)and metalloproteinase-9(MMP-9)].Results:Compared with that in the blank group,the inhibition rate of proliferation of SW480 cells increased significantly with the increase of pyrethrin concentration(5,10 and 15μmol·L^-1)within the detection range(P<0.05).Under the same concentration,the inhibition rate of proliferation increased significantly with the increase of action time(P<0.05).There was significant difference between parithenolide at 10 and 15μmol·L^-1 concentration groups and the positive control group.Compared with that in the blank control group,the numbers of clonal cells,invasion and migration cells,and protein levels of proliferation-related proteins beta-catenin and Cyclin D1,and invasion and migration-related proteins MMP-2,MMP-9 and NOX4 in pyrethrin groups decreased significantly(P<0.05),and there was a concentration-dependent relationship among parithenolide groups(P<0.05).Compared with the positive control group,there were significant differences in the above indices of pyrethrin at 10 and 15μmol·L-1 concentration groups(P<0.05).Conclusion:Parithenolide may inhibit the proliferation and invasion of colon cancer SW480 cells by down-regulating the expression of NOX4.