pCS2-MT/Axin及pEGFP-N3/APC重组质粒在结肠癌细胞系SW480中的表达  

Expression of recombinant plasmid pCS2-MT/Axin and pEGFP-N3/APC in SW480 colon carcinoma cell line

作　　者：徐玫丽[1] 霍继荣[1] 吕梁[1] 

机构地区：[1]中南大学湘雅二医院消化内科,湖南长沙410011

出　　处：《中国现代医学杂志》2009年第8期1139-1142,共4页China Journal of Modern Medicine

摘　　要：目的在结肠癌细胞系SW480中成功表达Axin和APC基因片段。方法双酶切鉴定重组质粒pCS2-MT/Axin及pEGFP-N3/APC5,利用脂质体介导法将质粒共转染结肠癌细胞系SW480,RT-PCR鉴定细胞中目的基因APC和Axin mRNA的表达,western blotting鉴定标签蛋白GFP和myc的表达。结果重组质粒双酶切得到的片段大小与预期相符,质粒共转染SW480后,RT-PCR检测到目的基因APC和AxinmRNA的表达,western blotting检测到标签蛋白GFP和myc的表达。结论重组质粒pCS2-MT/Axin及pEGFP-N3/APC5在结肠癌细胞中成功表达,为下一步研究APC和Axin基因在细胞内的功能奠定了实验基础。[Objective] To express Axin gene and APC gene fragment in colon cancer cell SW480. [Method] The recombinant plasmid pCS2-MT/Axin and pEGFP-N3/APC5 was identified by double digested. Plasmid was eotransfected into SW480 cell line by liposome. The target gene Axin and APC were identified by reverse transcription -polymerase chain reaction （RT-PCR）, and the label protein GFP and MYC were identified by western blotting. [Result] The agarose gel elctrophrosis showed a 4.3 kb and a 2.5 kb band of the digestion product of pCS2-MT/ Axin, a 4.7 kb and a 2.0 kb band of pEGFP-N3/APC5, which matched the expected size. The target genes can be detected by RT-PCR,and the label proteins can be detected by western blotting. [ Conclusion] The recombinant plasmid pCS2-MT/Axin and pEGFP-N3/APC5 could be expressed successfully in SW480 cell line .It may provide an experimental basis for the further study of their function in cells.

关 键 词：轴蛋白 结肠腺瘤性息肉病基因 结直肠癌 转染 

分 类 号：R735.3[医药卫生—肿瘤]