CAJ | 학술논문

目的探讨Livin基因和Survivin基因联合靶向小干扰(si)RNA对结肠癌细胞增殖及凋亡的影响.方法构建Livin和sunrivin联合靶向的siRNA重组表达载体并转染结肠癌细胞,通过RT-PCR和蛋白质印迹方法分别检测Livin和Survivin的表达.通过MTT法检测siRNA对细胞增殖的抑制作用.利用流式细胞仪检测处理后细胞的凋亡效应.结果经酶切鉴定和测序分析证实Liyin和Survivin联合靶向的siRNA重组表达载体构建成功.这种联合靶向SiRNA对Livin mRNA及蛋白表达的抑制率分别为27.9%和22.3%.对Survivin mRNA及蛋白表达的抑制率分别为32.2%和40.9%.与对照组相比,联合靶向siRNA可降低肿瘤细胞增殖率,增加细胞凋亡率,但其细胞生长抑制作用和促细胞凋亡作用均弱于单独干扰Livin或Survivin基因.结论 Livin和Survivin基因联合靶向siRNA能降低结肠癌细胞中Livin和Survivin基因的表达.抑制结肠癌细胞的增殖.并诱导结肠癌细胞的凋亡.但此协同抑制作用较单独干扰Livin或Survivin基因为弱.
목적 탐토Livin기인화Survivin기인연합파향소간우(si)RNA대결장암세포증식급조망적영향.방법 구건Livin화sunrivin연합파향적siRNA중조표체재체병전염결장암세포,통과RT-PCR화단백질인적방법분별검측Livin화Survivin적표체.통과MTT법검측siRNA대세포증식적억제작용.이용류식세포의검측처리후세포적조망효응.결과 경매절감정화측서분석증실Liyin화Survivin연합파향적siRNA중조표체재체구건성공.저충연합파향SiRNA대Livin mRNA급단백표체적억제솔분별위27.9%화22.3%.대Survivin mRNA급단백표체적억제솔분별위32.2%화40.9%.여대조조상비,연합파향siRNA가강저종류세포증식솔,증가세포조망솔,단기세포생장억제작용화촉세포조망작용균약우단독간우Livin혹Survivin기인.결론 Livin화Survivin기인연합파향siRNA능강저결장암세포중Livin화Survivin기인적표체.억제결장암세포적증식.병유도결장암세포적조망.단차협동억제작용교단독간우Livin혹Survivin기인위약.
Objective To investigate the effect of siRNA targeting Livin and Survivin gene simultaneously on the proliferation and apoptosis of human colon cancer cells. Methods SiRNA recombinant expression vectors targeting Livin and Survivin gene simultaneously were constructed andtransfected into human colon cancer cell line Lovo. The effects of siRNA recombinant expression vector on Lovo cells were detected by RT-PCR, Western blot, MTT reduction assay and flow cytometry.Results It was confirmed by restriction endonuclease and sequence analysis that siRNA recombinant expression vector targeting Livin and Survivin gene simultaneously was constructed successfully. The suppressive rates of siRNA targeting Livin and Survivin gene simultaneously on I.ivin mRNA and protein expression were 27.9% and 22.3% respectively, and those on Survivin mRNA and protein expression were 32.2% and 40.9% respectively. The survival rate of cancer cells was decreased whereas the apoptotic rate was increased, but the coordinate repression was weaker than Livin and Survivin RNA interference alone. Conclusions siRNA targeting Livin and Survivin gene simultaneously can decrease the expression of Livin and Survivin gene, suppress cell proliferation and induce cell apeptosis in human colon cancer. The coordinate repression was weaker than Livin and Survivin RNA interference alone.