Raf激酶抑制蛋白对鼻咽癌细胞增殖的影响

Effects of RKIP on the proliferation of nasopharyngeal carcinoma cells

目的:探讨Raf激酶抑制蛋白(RKIP)对鼻咽癌细胞增殖的影响。方法:采用脂质体转染方法将反义RKIP核酸表达质粒pcDNA3.1(-)-asRKIP及空白载体pcDNA3.1(-)分别转染鼻咽癌细胞系6-10B细胞,用West-ern印迹检测RKIP表达,建立RKIP表达下调的稳定转染细胞和对照细胞系。采用四甲基偶氮唑蓝(MTT)法、流式细胞术和软琼脂集落形成实验检测RKIP表达下调对鼻咽癌细胞增殖、细胞周期分布和停泊非依赖性生长的影响。结果:建立了RKIP表达下调的6-10B鼻咽癌细胞系;RKIP表达下调促进6-10B细胞增殖和停泊非依赖性生长,并加速其越过G0/G1期。结论:RKIP具有抑制鼻咽癌细胞增殖和停泊非依赖性生长的作用,可能在鼻咽癌发病中具有重要作用。

Objective To investigate the effect of Raf kinase inhibitor protein （RKIP） on the proliferation of nasopharyngeal carcinoma （NPC） cells. Methods pcDNA3.1 （ - ） -asRKIP plasmid that constitutively expressed antisense RKIP mRNA and empty plasmid pcDNA3.1 （ - ） were transfected into human NPC cell line 6-10B cells using Lipofectamine2000 to establish cell line With RKIP down-regulation and control cell line, respectively. Methyhhiazoletetrazolium （MTT） assay, flow cytometry （FCM） analysis and soft-agar colony formation assay were used to examine the effects of RKIP down-rgulation on proliferation, cell cycle distribution and anchorage independence growth of 6-10B cells, respectively. Resuits Stably transfected 6-10B cell line with RKIP down-regulation was successfully established. Downregulation of RKIP in 6-10B cells could promote cell proliferation and anchorage independence growth, and increase the number of G0/G1 phase cells. Conclusion RKIP could inhibit cell proliferation and anchorage independence growth of human NPC cells, suggesting that RKIP might be involved in the pathogenesis of NPC.