CRNDE过表达促进人肝癌细胞系HepG2增殖和迁移

目的探讨长链非编码RNA(lncRNA)结直肠差异性表达基因(CRNDE)对HepG2细胞增殖和迁移能力的影响及作用机制。方法构建CRNDE过表达质粒,进行慢病毒包装后感染HepG2细胞。实验分别设置阴性对照组(LV5/NC)和CRNDE过表达组(LV5/CRNDE)。应用2.5μg/mL嘌呤霉素干预4~5周,筛选出CRNDE过表达HepG2细胞系;CCK8和细胞划痕实验检测细胞的增殖及迁移能力变化;real-time PCR和Western blot检测两组细胞E-cadherin、N-cadherin、Bax和Bcl-2 mRNA及蛋白表达变化。结果与LV5/NC组相比,LV5/CRNDE组CRNDE表达显著升高(P<0.01),且LV5/CRNDE组细胞的增殖和迁移能力均显著提高(P<0.01);同时,LV5/CRNDE组细胞E-cadherin和Bax表达均明显降低(P<0.01),而N-cadherin和Bcl-2表达则明显升高(P<0.01)。结论CRNDE可通过促进N-cadherin和Bcl-2表达,抑制E-cadherin和Bax表达,进而增强HepG2细胞的增殖和迁移能力。

Over-expression of CRNDE promotes proliferation and migration in human hepatoma cell line HepG2

Objective To investigate the effects and mechanisms of long-chain non-coding RNA CRNDE on the proliferation and migration of HepG2 cells.Methods The CRNDE over-expression vector was constructed and packed into lentivirus.Then,HepG2 cells was infected by the lentivirus containing LV5/NC and LV5/CRNDE vectors.After treatment with 2.5μg/mL puromycin for 4-5 weeks,real-time PCR was used to detect the expression level of CRNDE in the two groups.CCK8 and cell scratch assay was used to detect the changes of proliferation and migration ability of each group.Meanwhile,real-time PCR and Western blot were used to detect the mRNA and protein expression of E-cadherin,N-cadherin,Bax and Bcl-2 in each group.Results Compared with LV5/NC group,the expression of CRNDE in LV5/CRNDE group significantly increased(P<0.01).The proliferation and migration of LV5/CRNDE group cells were also significantly increased(P<0.01,P<0.01).At the same time,the expression of E-cadherin and Bax in LV5/CRNDE group significantly decreased(P<0.01,P<0.01),while the expression of N-cadherin and Bcl-2 significantly increased(P<0.01,P<0.01).Conclusions CRNDE may stimulate proliferation and migration of HepG2 cells by promoting the expression of N-cadherin/Bcl-2 and inhibiting the expression of E-cadherin and Bax.