敲减Akt表达对类风湿关节炎成纤维样滑膜细胞增殖的影响

Effects of down-regulating of Akt protein on the proliferation of rheumatoid arthritis fibroblast-like synoviocytes

目的:研究慢病毒介导Akt靶向的RNA干扰对类风湿关节炎成纤维样滑膜细胞(RAFLS)增殖的影响。方法:慢病毒表达载体pL/Akt转染RAFLS并表达靶向Akt的siRNA序列,采用RT-PCR和Western Blot检测Akt表达,MTT法检测RAFLS增殖能力的变化。结果:转染pL/Akt后,RAFLS中Akt1、Akt2的mRNA分别下降了65%和68%以上,Akt蛋白的表达量在48h后下降76.19%(P<0.05),RAFLS细胞增殖在48、72、96h分别下降了27.50%、44.69%和56.80%(P<0.05)。结论:慢病毒表达载体pL/Akt能够降低Akt蛋白表达水平,并显著抑制RAFLS增殖。

Objective To study the effects of lentiviral vector-mediated small interfering RNA targeting Akt protein on the proliferation of rheumatoid arthritis （RA） fibroblast-like synoviocytes （FLS）. Methods Small interfering RNA targeting Akt protein in RA FLS was expressed, and then the expression of Akt was detected by RT-PCR and Western Blot, and the proliferation of RA FLS was measured by MTT method. Results The expressions of Aktl and Akt2 mRNA were decreased by more than 65% and 68% respectively in RA FLS after being transfected with lentiviral vector pL/Akt. The expression of Akt protein was reduced by 76.19% 48 h later （P 〈 0.05）, and the proliferation of RA FLS was decreased by 27.50%, 44.69%, and 56.80% （P 〈 0.05） at 48 h, 72 h, and 96 h later. Conclusion Lentiviral vector pL/Akt could decrease the expression of Akt protein, and significantly inhibit RA FLS proliferation.