萝卜硫素对结肠癌SW480细胞增殖、侵袭及Notch通路的影响

Effects of sulforaphane on proliferation,invasion and Notch pathway of colorectal cancer SW480 cells

目的研究萝卜硫素(SFN)对人结肠癌SW480细胞增殖、侵袭及Notch通路的影响。方法MTT法测定不同浓度SFN对SW480细胞生长抑制作用,Transwell侵袭实验测定1、2、5μmol/L SFN对SW480细胞体外侵袭能力的影响,划痕实验测定1、2、5μmol/L SFN对SW480细胞体外迁移能力的影响,Western blotting法测定1、2、5μmol/L SFN处理后SW480细胞Notch、Hes1、Ki-67、增殖细胞核抗原(PCNA)、基质金属蛋白酶9(MMP-9)、E-cadherin蛋白表达水平。结果与对照组比较,1、2、5、10、20、40μmol/L SFN处理后均能显著抑制SW480细胞增殖(P<0.05)。5μmol/L SFN作用48 h后对SW480细胞抑制率为(26.38±3.24)%,细胞活力大于70%,为防止SW480细胞活力过低导致侵袭实验和划痕实验出现假阳性结果,因此后续实验选取SFN浓度1、2、5μmol/L进行。侵袭和迁移实验发现,与对照组比较,人结肠癌SW480细胞经1、2、5μmol/L SFN处理48 h后侵袭能力、迁移能力、Ki-67、PCNA、MMP-9蛋白表达水平显著降低(P<0.05),E-cadherin蛋白表达水平显著升高(P<0.05),呈浓度依赖性;与对照组比较,人结肠癌SW480细胞经1、2、5μmol/L SFN处理48 h后,Notch、Hes1蛋白表达水平显著降低(P<0.05)。结论SFN可能通过阻断Notch通路活化,下调Hes1表达水平,达到抑制结肠癌SW480细胞增殖、迁移、侵袭的目的。

Objective To study the effects of sulforaphane(SFN)on the proliferation,invasion and Notch pathway of human colorectal cancer SW480 cells.Methods MTT assay was used to determine the inhibitory effects of SFN at different concentrations on the growth of SW480 cells,Transwell invasion assay was used to determine the effects of 1,2 and 5μmol/L SFN on the invasion ability of SW480 cells in vitro,scratch assay was used to determine the effects of 1,2 and 5μmol/L SFN on migration of SW480 cells in vitro,the expression levels of notch,Hes1,Ki-67,PCNA,MMP-9 and E-cadherin in SW480 cells treated with 1,2 and 5μmol/L SFN were determined by Western blotting.Results Compared with the control group,1,2,5,10,20 and 40μmol/L SFN could significantly inhibit the proliferation of SW480 cells(P<0.05).The inhibition rate of 5μmol/L SFN on SW480 cells was(26.38±3.24)%and the cell viability was more than 70%.In order to prevent false positive results in the invasion and scratch experiment,the concentrations of 1,2 and 5μmol/L SFN were selected for subsequent experiments.Invasion and migration experiments showed that,compared with control group,after the human colorectal cancer SW480 cells treated with 1,2 and 5μmol/L SFN for 48 h,the expression levels of invasion,migration,Ki-67,PCNA,MMP-9 proteins decreased significantly(P<0.05),and the expression of E-cadherin protein increased significantly(P<0.05),in a concentration-dependent manner;compared with the control group,the expressions of Notch and Hes1 proteins in human colon cancer SW480 cells decreased significantly after treating with 1,2 and 5μmol/L SFN for 48 hours(P<0.05).Conclusion SFN may inhibit the proliferation,migration and invasion of colon cancer SW480 cells by blocking Notch pathway activation and lowering Hes1 level.