全反式维甲酸对白血病NB4细胞PRAM蛋白的影响

EXPRESSION OF PRAM PROTEIN IN MYELOID LEUKEMIA NB4 CELLS INFLUENCED BY ATRA

目的:探讨在NB4细胞增殖过程中PRAM蛋白表达情况,并且用全反式维甲酸(ATRA)进行干扰,观察其对PRAM蛋白的影响。方法:①细胞增殖抑制实验:体外培养NB4细胞共5天,用台盼蓝染色法筛选ATRA有效干预浓度后,选取ATRA(1×10^(-6)mol/L)为目的干预浓度。②Western blot:ATRA(1×10^(-6)mol/L)持续干扰NB4细胞1、2、3天后检测PRAM蛋白的相对表达量。结果:①与对照组比较,4种浓度ATRA组细胞与NB4细胞的增殖率呈量效与时效关系;浓度越大,抑制率越高;于培养第4天出现最强抑制。②选取1×10^(-6)mol/L ATRA干扰NB4细胞共3天,对照组及ATRA组PRAM蛋白的表达在时间上有规律性:第2天表达最强烈。③1×10^(-6)mol/L ATRA使PRAM蛋白在第2天的相对表达量较对照组低(P<0.05)。④1×10^(-6)mol/l ATRA使NB4细胞PRAM蛋白在第2天出现最大表达量,同时该组细胞出现生长抑制。结论:ATRA能通过PRAM位点来达到抑制APL增殖分化的目的。

Objective： To discuss the expression of protein PRAM （ PML-RARa target gene encoding an Adaptor Molecule, PRAM） in myeloid leukemia cells NB4 influenced by ATRA. Methods： NB4 cell line was cul- tured in 4 different concentrations of ATRA, and cells proliferation were detected .Then ATRA （1×10^-6molJL） waschosen to add into cell culture, 1,2,3 days later, PRAM protein were tested by Western blot. Results： ①In the process＇of NB4 cells cultured in vitro, the expression of PRAM protein was down-regulated by 10^-6mol/L ATRA compared with control group, especially on the 2nd day.②Protein PRAM expressed regularly, which was higher on the 2nd day and lower on the 3rd day. Conclusions： ①1×10^-6mol/L ATRA can attenuate NB4 cells to mature.② ATRA leads to decreased expression of PRAM protein.It suggess that differentiation of acute promyelocytic leukemia cells is disrupted during transformation by PML-RARct and induced by ATRA signaling events .