刘冀龙1 李彦娟2 刘继光3 李岩3 董新舒4.封闭EFNA2 基因表达对肝癌细胞HepG2 的转移和运动的抑制作用[J].,2012,12(25):4841-4844 |
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 LiverCancer Cell Line HepG2 |
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DOI: |
中文关键词: EFNA2 重组腺病毒 肝癌HepG2 细胞 侵袭和运动 |
英文关键词: EFNA2 siRNA Reconstruction adenovirus Liver cancer HepG2 cell Invasion and motor |
基金项目:国家自然科学基金资助项目(30170828) |
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中文摘要: |
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。 |
英文摘要: |
Objective: To reconstruct Ephrin-A2(EFNA2) siRNA adenovirus vector and to observe its effect on the metastasis ability
of liver cancer cell line HepG2. Methods: EFNA2 siRNA was constructed into adenovirus vector pAD-X by gene recombination technique,
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 reconstruction
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 reconstructed
and it could inhibit invasion and motor abilities of transfected HepG2 cells. |
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