稳定沉默黏着斑激酶结肠癌细胞株SW620的构建

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摘要目的：构建稳定沉默黏着斑激酶（FAK）结肠癌细胞株SW620。方法：用构建好的FAK RNA干扰（RNAi）慢病毒载体pLL3.7 FAK及插入无沉默功能序列的阴性对照慢病毒载体pLL3.7，在293FT细胞中包装成慢病毒，感染体外培养结肠癌SW620细胞，作为实验组和阴性对照组，未感染病毒的SW620细胞作为未处理组，通过荧光倒置显微镜检测SW620细胞的感染效率，运用RT-PCR、Western blot方法测定FAK的表达情况，建立稳定转染细胞株。结果：成功包装了靶向FAK的RNAi慢病毒，感染效率＞90%，稳定转染细胞株构建成功。RT-PCR及Western blot方法检测发现实验组与阴性对照组及未处理组相比，FAK的表达明显降低。结论：稳定沉默FAK的RNAi SW620细胞构建成功。

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	洪卫文1
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关键词 ：黏着斑激酶, RNA干扰, SW620, 结直肠肿瘤, 转移

Abstract：Objective: To establish the FAK stable silencing SW620 cells. Methods: For production of lentiviruses, 293FT cells were transfected by Turbo transfect, using pLL3.7 FAK and negative pLL3.7, lentivirus-containing media were harvested and used to infect SW620 cells as experimental group and control group, SW620 cells didn抰 infect any lentiviruses as unsettled group. Infection efficiency was obtained by the expression of GFP, and maintained as a pool population. Expression of FAK was detected by RT-PCR and Western blot. Results: Lentiviruses were successfully producted with over 90% infecting rate; cells stable expressing GFP. Expression of FAK were significantly reduced by RT-PCR and Western blot. Conclusion: FAK stable silencing SW620 cells are established.

Key words： focal adhesion kinase RNA interference SW620 colorectal neoplasms metastasis

收稿日期: 2013-03-18

作者简介: 洪卫文（1983-），男，福建龙岩人，住院医师，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2014/V44/I3/201

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