鸟氨酸脱羧酶基因反义RNA对淋巴瘤细胞Jurkat生长的影响

Effect of Antisense RNA of Ornithine Decarboxylase Gene on Jurkat Cell

探讨鸟氨酸脱羧酶(ornithine decarboxylase,ODC)反义RNA是否对人淋巴瘤细胞Jurkat的生长具有抑制作用。含反义RNA的真核表达载体pcDNA3.1(+)/Rodc用脂质体转染Jurkat细胞,G418筛选ODC表达抑制的细胞株,MTS法分析细胞增殖,Western blotting检测细胞中ODC蛋白表达水平,半定量RT-PCR检测细胞中ODC mRNA含量,流式细胞术检测细胞周期的变化,DNA片段化分析细胞凋亡。结果显示,成功获得ODC表达抑制的淋巴瘤细胞株J/o,ODC反义RNA转染细胞后,引起Jurkat细胞生长缓慢和S/G2细胞周期停滞,细胞对抗癌药物DFMO敏感性显著增加。由此证明,ODC反义RNA能抑制人T淋巴瘤Jurkat细胞的生长,具有治疗人白血病的潜在应用价值。

The study was designeol to explore the effects of antisense RNA of ornithine decarboxylase （ODC） on Jurkat cells. The eukaryotie expression plasmid for ODC antisense RNA was constructed. The plasmid was then transfeeted into Jurkat cells with lipofectamine reagent. The cell line with ODC expression inhibition was selected by RT-PCR and Western blotting assay, Using this cell line as a model, the effects of ODC antisense RNA on cell growth and cell shap（ MTS assay） , cell cycle （flow cytometry assay）and cell sensitive to DFMO （ MTS and DNA fragmentation assay） were determined. Results indicated that the plasmid pcDNA3.1 （ ＋ ）/Rode for ODC antisense RNA was transfected into Jurkat cells successfully and a Jurkat cell line （J/o）with stable low-expression of ODC was obtained. Comparing with the control cells,low-expression of ODC in Hrl cell line resulted in growth inhibition and cell shape change, cell cycle arrest at G2 phase and increased sensitivity to antitumor drug DFMO. Therefore,it proved that Antisense RNA of ODC can inhibit growth of Jurkat cells and has potential in the anti-tumor therapy.