长链非编码RNA EVADR促进人结直肠癌细胞系增殖及迁移

摘要

目的 探讨IncRNA EVADR对人结直肠癌HCT116和LOVO细胞系的增殖和迁移的影响以及其作用机制.方法 利用慢病毒感染方式分别构建稳定 过表达IncRNA EVADR的HCT116和LOVO细胞系;用CCK-8检测细胞的增殖;Transwell小室法检测细胞迁移能力,Western blot检测E-cadherin的表达, 实时荧光定量PCR检测Snail、 Slug、ZEBl和ZEB2 mRNA的表达.结果 成功构建了稳定过表达IncRNA EVADR的结直肠癌HCT116和LOVO细胞系.与 HCT116-NC和LOVO-NC细胞组对比,HCT116-EVADR与LOVO-EVADR细胞组的增殖和迁移能力明显提高(P＜0.05).同时,上皮标志物E-cadherin表达明显降 低,而Snail、Slug、ZEB1和ZEB2间质化标志物的表达升高.结论 过表达IncRNA EVADR具有促进结直肠癌HCT116和LOVO细胞的增殖与迁移能力,可能 通过调节结直肠癌细胞上皮间质转化(EMT)发挥作用.%Objective To explore the effect and mechanism of IncRNA EVADR on the proliferation and migration in human colorectal cancer cell lines HCT116 and LOVO. Methods HCT116 and LOVO cell lines were transfected with IncRNA EVADR by overexpressing lentivirus system. CCK8 assay was performed to measure the growth of HCT116 and LOVO cells after overexpression of EVADR. Transwell migration was performed to determine if EVADR promote HCT116 and LOVO cells migration. Finally, the expression of Ecadherin and transcription factor Snail, Slug, ZEB1 and ZEB2 were detected by Western blot and realtime quantitative PCR respectively. Results We successfully established colorectal cancer cells strains HCT116, LOVO which can stably overexpress IncRNA EVADR and the capacity of proliferation and migration in overexpression group was significantly improved (P＜0.05). The expression of Ecadherin was decreased while mesenchymal markers Snail, Slug, ZEB1 and ZEB2 were increased in EVADR overexpression HCT116 and LOVO cells. Conclusions Overexpression of IncRNA EVADR in HCT116 and LOVO cells can significantly promote the proliferation and migration of HCT116 and LOVO cells which may play an important role in regulating epithelial-mesenchymal transition in colorectal cancer cells.