阻断Notch信号通路对肝癌细胞迁移和COX-2蛋白表达的影响

摘要

目的 探讨阻断Notch信号通路对肝癌细胞迁移的和环氧化酶-2(COX-2)蛋白表达的影响.方法 在体外培养肝癌细胞系HepG2、Huh-7和正常非肿瘤细胞系HL-7702,在Transwell小室检测细胞的迁移能力,对不同肝癌细胞系及正常非肿瘤肝细胞系的侵袭迁移能力进行比较;采用Western Blot的方法检测COX-2、E-cadher-in、Snaill的蛋白表达;采用5μmol/L的 γ-分泌酶抑制剂(DAPT)阻断Notch信号通路,通过对肝癌细胞系HepG2、Huh-7和对照组(空培养基)细胞迁移侵袭的比较得出DAPT和NS-398阻断Notch信号通路对肝癌细胞迁移能力的影响.结果 不同肝癌细胞系及正常非肿瘤肝细胞系的侵袭迁移能力比较结果显示,HL-7702细胞系细胞的迁移和侵袭数量分别为(128.23±10.34)、(112.34±9.02)个,低于HepG2、Huh-7细胞系细胞迁移、侵袭数量,差异有统计学意义(P<0.05);DAPT和NS-398处理对肝癌细胞迁移能力的影响结果显示,加5μmol/L的DAPT后HepG2的细胞迁移侵袭数量分别为(213.36±9.02)和(197.53±11.27)个,Huh-7的细胞迁移侵袭数量分别为(201.74±11.23)和(179.63±9.26)个,采用50μmol/L NS-398后,HepG2的细胞迁移侵袭数量分别为(203.26±11.25)和(187.54±11.37)个,Huh-7的细胞迁移侵袭数量分别为(187.63±8.87)和(165.64±8.45)个.与对照组相比,使用DAPT或NS-398处理后的肝癌细胞在Transwell小室中迁移侵袭能力均显著降低,两者比较差异有统计学意义(P<0.05);HepG2和Huh-7两细胞系在5μmol/L DAPT干预后,COX-2、Snail蛋白表达明显出现下调,而E-cadher-inl表达明显出现上调现象.结论 Notch信号通路在肝癌细胞侵袭迁移过程中起着非常重要的作用,阻断后细胞的迁移能力下降,Notch调控COX-2后其蛋白表达下调,然后调控Snail/E-cadherin的表达,从而改变肿瘤细胞的迁移过程.%Objective To explore effects of blocking Notch signaling pathway on cell migration and expression of COX - 2 protein in hep-atocellular carcinoma cells. Methods The hepatocellular carcinoma cell line HepG2,Huh - 7 and normal non tumor cell line HL - 7702 were in vitro cultured,Transwell cell migration ability of hepatoma cell lines and normal non tumor liver cell line invasion and migration ability were com-pared. Western blot method was performed for the detection of snail and E - cadherin and cyclooxygenase 2 (COX - 2),E - cadher - in,Snaill-protein expression;using 5 μmol/ L of dapt gamma secretase inhibitor blocking Notch signaling pathway,based on human hepatoma cell line HepG2,Huh - 7 and control group (empty medium)cell migration and invasion of dapt. Results The invasion and migration of hepatoma cell lines and normal non tumor liver cell line comparison results showed that the number of migration and invasion of HL - 7702 cells were (128. 23 ± 10. 34),(112. 34 ± 9. 02),HepG2 and Huh - 7 cell lines,cell migration,invasion was statistically significant compared to the number difference (P < 0. 05 DAPT;NS)and the effect of - 398 treatment on liver cancer cell migration results showed that adding 5 μmol/ L DAPT after HepG2 cell migration and invasion were (213. 36 ± 9. 02)and (197. 53 ± 11. 27),Huh - 7 cell migration and invasion were (201. 74 ± 11. 23)and (179. 63 ±9. 26),using 50 μmol/ L NS -398,HepG2 cell migration and invasion were (203. 26 ±11. 25)and (187. 54 ± 11. 37),Huh - 7cell migration and invasion were (187. 63 ±8. 87)and (165. 64 ±8. 45),compared with the control group,the dapt or ns - 398 treatment ofhepatoma cells in Transwell migration invasion were significantly lower,and the difference was statistically significant (P <0. 05). HepG2,Huh-7 cells in 5 μmol/ L dapt intervention,snail and COX -2 protein expression was significantly down regulated,and E - cadher - inl expressionsignificantly increased phenomenon. Conclusion Notch signaling pathway in hepatocellular carcinoma cell invasion plays a very important role inthe migration process,after the occlusion of the decline in the ability of cell migration. Notch can regulate COX -2 and protein down - regulated,and regulate Snail/ E - cadherin expression,so as to change the tumor cell migration.