| Skp2 shRNA表达质粒的构建及其对肺癌细胞生长的影响 |
| |
| 引用本文： | 李胜,梅同华,黎联,张明川.Skp2 shRNA表达质粒的构建及其对肺癌细胞生长的影响[J].第三军医大学学报,2008,30(13):1275-1278. |
| |
| 作者姓名： | 李胜 梅同华 黎联 张明川 |
| |
| 作者单位： | 重庆医科大学附属第二医院呼吸内科,重庆,400010 |
| |
| 摘 要： | 目的 构建Skp2基因的RNAi真核表达质粒,观察其对SIC-A-1肺癌细胞内Skp2基因表达及细胞生长的影响.方法 根据GenBank中Skp2cDNA序列设计针对Skp2基因的shRNA序列,合成序列并克隆人pGenSil-1质粒中,构建skp2 pshRNA重组质粒.脂质体介导重组质粒转染SPC-A-1肺癌细胞.RT-PCR检测肺癌细胞Skp2 mRNA的表达,Western blot检测Skp2蛋白表达.MTT检测各纽细胞生长情况,流式细胞仪检测细胞周期改变.结果 重组质粒经酶切鉴定和测序,证实表达干扰质粒构建成功.转染重组质粒的肺癌细胞Skp2 mRNA和蛋白的表达明显下降(P<0.05),细胞生长增殖受到抑制,阻滞在G0/G1期的细胞比例增加,而进入S期的细胞比例明显下降(P<0.05).结论 构建的Skp2 shRNA表达质粒能有效下调Skp2基因的mRNA及蛋白的表达、抑制细胞的生长增殖.
|
| 关 键 词： | Skp2基因 RNA干扰 短发夹RNA 肺癌细胞 |
Construction of Skp2 shRNA and its effect on lung cancer cell growth |
| |
| LI Sheng,MEI Tong-hua,LI Lian,ZHANG Ming-chuan.Construction of Skp2 shRNA and its effect on lung cancer cell growth[J].Acta Academiae Medicinae Militaris Tertiae,2008,30(13):1275-1278. |
| |
| Authors: | LI Sheng MEI Tong-hua LI Lian ZHANG Ming-chuan |
| |
| Institution: | LI Sheng,MEI Tong-hua,LI Lian,ZHANG Ming-chuan(Department of Respiratory Diseases,The Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China) |
| |
| Abstract: | Objective To construct and identify the RNAi eukaryotic vector of Skp2 gene and to observe its interfering effect on the growth of SPC-A-1 lung cancer cells.Methods The specific shRNA sequence was designed and synthesized according to the Skp2 cDNA sequence in GenBank.The sequence was cloned into plasmid pGenesil-1.Then recombinant vector was transfected into SPC-A-1 lung cancer cells by Lipofectamine 2000.The expressions of Skp2 mRNA were analyzed by RT-PCR and the levels of Skp2 protein were detected by W... |
| |
| Keywords: | Skp2 gene RNA interference small hairpin RNA lung cancer cell |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
