类泛素化修饰抑制剂对人胃癌细胞增殖和迁移的影响

Effects of neddylation modification inhibitor MLN4924 on proliferation and migration of human gastric cancer cells

目的观察NEDD8活化酶(NAE)抑制剂MLN4924对人胃癌细胞增殖和迁移的影响。方法使用不同浓度MLN4924处理人胃癌细胞株NCI-N87和SGC-7901细胞72 h,细胞计数试剂盒(CCK-8)法检测抑制率。选取20%抑制浓度(IC20)浓度处理NCI-N87和SGC-7901细胞,CCK-8法和克隆形成实验检测细胞增殖能力的变化,细胞划痕实验检测细胞迁移能力的变化。结果当MLN4924浓度分别达到0.528 μmol/L和0.375 μmol/L时,胃癌细胞NCI87和SGC-7901的增殖被抑制20%。CCK-8法检测结果显示,MLN4924能明显抑制胃癌细胞的增殖,且抑制作用具有剂量和时间依赖性。克隆形成实验结果表明NCI-87细胞经MLN4924作用后,克隆数低于对照组[(71.00±10.19)个比(215.33±9.56)个,P<0.01]。SGC-7901细胞经MLN4924作用后,克隆数低于对照组[(23.66±18.66)个比(132.33±34.12)个,P<0.01]。划痕实验结果显示,NCI-87细胞划痕培养48 h后,对照组细胞面积愈合率为(51.05±6.35)%,MLN4924作用后面积愈合率为(5.30±0.78)%。SGC-7901细胞划痕培养48 h后,对照组细胞面积愈合率为(23.05±2.01)%,MLN4924作用后面积愈合率为(3.12±0.52)%。结论类泛素化修饰抑制剂MLN4924可抑制人胃癌细胞的增殖和迁移。

Objective To explore the effects of NEDD8 activating enzyme (NAE) inhibitor MLN4924 on gastric cancer cell proliferation and migration. Methods Human gastric cancer cell lines NCI-N87 and SGC-7901 were treated with different concentrations of MLN4924, and the inhibitory concentrations were detected by cell counting kit-8 (CCK-8) method. 20% maximal inhibitory concentration (IC20) was used to treat gastric cancer cells. The cell proliferation was measured by CCK-8 and clone formation assay, and cell migration was determined by cell scratch test. Results When human gastric cancer cell lines NCI-N87 and SGC-7901 were treated with 0.528 and 0.375 μmol/L respectively, the inhibitory rate reached to 20%. CCK-8 method showed that MLN4924 inhibited the proliferation of human gastric cells in a dose- and time-dependent manner. In the clone formation assay, the clone number of NCI-N87 cells was 71.00±10.19 in MLN4924 treated group and 215.33±9.56 in the control group. The clone number of SGC-7901 cells was 23.66±18.66 in MLN4924-treated group, and 32.33±34.12 in the control group.In vitro wound scratch assay revealed that the healing rate of MLN4924-treated NCI-N87 cells was (5.30±0.78)%, and (51.05±6.35)% in the control group, and that of MLN4924-treated SGC-7901 cells was (3.12±0.52)%, and (23.05±2.01)% in the control group. Conclusion Neddylation modification inhibitor MLN4924 suppresses the growth and migration of gastric cancer cells.