CAJ | 학술논문

目的：探讨受体相互作用蛋白140（ RIP140）过表达对小鼠神经母细胞瘤细胞（ N2a细胞）迁移和增殖的影响。方法采用脂质体转染法将RIP140过表达质粒导入N2a细胞，经G418筛选出稳定过表达RIP140的细胞系，利用实时定量PCR及Western 印迹法对构建的细胞模型进行鉴定；分别采用Transwell小室和活细胞计数试剂盒（CCK）8检测RIP140过表达对N2a细胞迁移和增殖的影响。结果将RIP140过表达质粒导入N2a细胞，成功构建RIP140过表达的 N2a细胞模型N2a-rip140，与N2a细胞相比，N2a-rip140细胞中RIP140 mRNA及蛋白表达量均增高。迁移检测显示N2a细胞迁移数明显高于N2a-rip140细胞[（70±20）个比（5±4）个， P＜0.05]。细胞增殖检测显示N2a细胞在24、48、72 h的增殖率分别为1.567±0.107、3.018±0.212、4.112±0.221，而N2a-rip140细胞为1.561±0.281、2.232±0.235、4.025±0.217，两种细胞在3个时间点的增殖率差异均无统计学意义（均P＞0.05）。结论 RIP140过表达抑制N2a细胞的迁移行为，但不影响其增殖行为。
목적：탐토수체상호작용단백140（ RIP140）과표체대소서신경모세포류세포（ N2a세포）천이화증식적영향。방법채용지질체전염법장RIP140과표체질립도입N2a세포，경G418사선출은정과표체RIP140적세포계，이용실시정량PCR급Western 인적법대구건적세포모형진행감정；분별채용Transwell소실화활세포계수시제합（CCK）8검측RIP140과표체대N2a세포천이화증식적영향。결과장RIP140과표체질립도입N2a세포，성공구건RIP140과표체적 N2a세포모형N2a-rip140，여N2a세포상비，N2a-rip140세포중RIP140 mRNA급단백표체량균증고。천이검측현시N2a세포천이수명현고우N2a-rip140세포[（70±20）개비（5±4）개， P＜0.05]。세포증식검측현시N2a세포재24、48、72 h적증식솔분별위1.567±0.107、3.018±0.212、4.112±0.221，이N2a-rip140세포위1.561±0.281、2.232±0.235、4.025±0.217，량충세포재3개시간점적증식솔차이균무통계학의의（균P＞0.05）。결론 RIP140과표체억제N2a세포적천이행위，단불영향기증식행위。
Objective To explore the effects of receptor interacting protein (RIP)140 gene over-expression upon the migration and proliferation of neuroblastoma cells in vitro.Methods The N2a RIP140 over-expression model (N2a-rip140) was constructed by lipofection and validated by real-time polymerase chain reaction ( PCR) and Western blot.The migration and proliferation capabilities were compared between N2a-rip140 and its parents by Transwell chamber and CCK-8.Results The N2a RIP140 over-expression model(N2a-rip140)was successfully constructed.Compared to N2a group, the RIP140 mRNA and protein expression levels of N2a-rip140 were remarkably up-regulated.Transwell assay showed that the over-expression of RIP140 inhibited N2a cell migration ((70 ±20) vs (5 ±4) cells, P<0.05).CCK-8 assay showed that the proliferation rates of N 2a group at 24, 48, 72 h were 1.567 ±0.107, 3.018 ±0.212 and 4.112 ±0.221 respectively.And those of N2a-rip140 group were 1.561 ±0.281, 2.232 ±0.235 and 4.025 ±0.217 respectively.No significant difference existed in proliferation rates at different timepoints among N2a, N2a-M and N2a-rip140 groups ( all P >0.05 ).Conclusion RIP140 over-expression effectively inhibits N2a cell migration.However it has no significant effect on the proliferation of N 2a cells.