封闭EFNA2基因表达对肝癌细胞HepG2的转移和运动的抑制作用  

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

作　　者：刘冀龙[1] 李彦娟[2] 刘继光[3] 李岩[3] 董新舒[4] 

机构地区：[1]北京市垂杨柳医院普外科,北京100022 [2]北京友谊医院超声科,北京100050 [3]佳木斯大学,黑龙江佳木斯154007 [4]哈尔滨医科大学附属第三临床医院腹部外科,黑龙江哈尔滨150001

出　　处：《现代生物医学进展》2012年第25期4841-4844,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金资助项目(30170828)

摘　　要：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。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,P0.01）.Conclusion： Recombinant adenovirus was correctly recon-structed and it could inhibit invasion and motor abilities of transfected HepG2 cells.

关 键 词：EFNA2 重组腺病毒 肝癌HEPG2细胞 侵袭和运动 

分 类 号：R735.7[医药卫生—肿瘤]