| 王红艳,涂永生,方 茅.人miR-29b过表达慢病毒载体的构建及其对A549细胞迁移的影响[J].中国肿瘤,2015,24(7):593-597. |
| 人miR-29b过表达慢病毒载体的构建及其对A549细胞迁移的影响 |
| To Construct Lentiviral Vector with Over-expressing miR-29b and To Explore Its Effect on Cell Migration in A549 Cell |
| 投稿时间：2015-01-07 |
| DOI：10.11735/j.issn.1004-0242.2015.07.A012 |
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| 中文关键词: miR-29b 慢病毒载体 细胞迁移 |
| 英文关键词:miR-29b lentiviral vector migration |
| 基金项目:广东省医学科研基金（A2014278）；广州市属高校科研项目（2012C135；2012C202）；广州医科大学博士启动科研项目（2014C08；2012C14） |
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| 中文摘要: |
| 摘 要：［目的］ 构建能高效表达人miR-29b的慢病毒载体，研究其对人肺腺癌A549细胞迁移的影响。［方法］ PCR扩增miR-29b前体序列，克隆入慢病毒表达载体，测序鉴定。包装好的慢病毒感染A549细胞，流式细胞仪分选后用荧光定量PCR检测miR-29b表达水平，划痕实验观察过表达miR-29b对A549细胞迁移的影响。［结果］ 测序结果证明成功构建人miR-29b过表达慢病毒载体。miR-29b过表达慢病毒感染效率为88.56%，使A549细胞中miR-29b表达升高2.78倍。与A549细胞相比，稳定过表达miR-29b的A549-miR-29b细胞的迁移能力显著性降低。［结论］ 成功构建miR-29b过表达慢病毒载体，过表达miR-29b可抑制A549细胞的迁移，为进一步研究miRNA对肿瘤转移的调控作用奠定了基础。 |
| 英文摘要: |
| Abstract：［Purpose］ To construct the lentiviral vector with overexpressing miR-29b，and to explore its effect on migration in human lung adenocarcinama cells A549. ［Methods］ The precursor of miR-29b was amplified by PCR and inserted into a lentiviral vector and sequenced. The recombinant lentiviral vector was further packaged for generating lentiviral to infect A549 cells. Stable miR-29b overexpressed A549-miR-29b cells were sorted by flow cytometry and detected the expression of miR-29b by real-time PCR. Wound healing assay was used to evaluate the effect of overexpressed miR-29b on cell migration of A549 cells. ［Results］ DNA sequencing demonstrated that miR-29b lentiviral vector was successfully constructed. The transduction efficiency in human A549 cells reached a level of 88.56%. The expression level of miR-29b increased 2.78 folds in infected A549 cell. Compared to A549 cell group，the migration abilities of A549-miR-29b cells significantly decreased. ［Conclusion］ The miR-29b lentiviral vector is constructed successfully. Overexpression of miR-29b could suppress migration abilities in A549 cell. It constructs the foundation of miRNA for regulation in the further research on tumor metastasis. |
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