封闭EFNA2 基因表达对肝癌细胞HepG2 的转移和运动的抑制作用

目的:构建Ephrin-A2(EFNA2)基因的干扰RNA重组腺病毒,并观察其对肝癌细胞HepG2转移能力的影响。方法:合成EFNA2基因的干扰RNA片段,用基因重组技术构建于腺病毒载体pAD-X,经293细胞包装得到高滴度重组腺病毒pAEFNA2/siRNA,并用real-time PCR进行EFNA2基因表达的验证。将重组腺病毒pAEFNA2/siRNA感染HepG2细胞,48h后用transwell小室法检测病毒对HepG2细胞侵袭和运动能力的影响。结果:经验证病毒正确构建,且HepG2细胞感染病毒pAEFNA2/siRNA后48小时的real-time PCR检测结果未见到有EFNA2基因的表达。病毒感染后transwell小室可见,HepG2细胞的侵袭和运动能力均受到显著的抑制(分别为105±12 v.s.21±7cells/孔和194±22 v.s.39±11,P<0.01)。结论:成功构建了重组EFNA2siRNA腺病毒,并观察到封闭EFNA2对肝癌细胞HepG2的侵袭和运动均有抑制作用。

Blocking EFNA2 Expression Inhibits Metastasis Ability of Human Liver Cancer Cell Line HepG2

Objective: To reconstruct Ephrin-A2(EFNA2) siRNA adenovirus vector and to observe its effect on the metastasis abil-ity of liver cancer cell line HepG2.Methods: EFNA2 siRNA was constructed into adenovirus vector pAD-X by gene recombination tech-nique,which was transformed into 293 packaged cell for high titer adenovirus.Real-time PCR was applied to detect EFNA2 siRNA gene expression.Transwell cabin assay was used to observe changes of invasion and motor ability of HepG2 cells transfected with reconstruc-tion adenovirus.Results: The finding of digestion was coincided with expected.No EFNA2 gene expression was detected in transfected HepG2 cells with real-time PCR.And the invasion and motor abilities of transfected HepG2 cells were significantly inhibited in transwell cabin assay(105±12 v.s.21±7cells/well,194±22 v.s.39±11,P<0.01).Conclusion: Recombinant adenovirus was correctly recon-structed and it could inhibit invasion and motor abilities of transfected HepG2 cells.