鸟氨酸脱羧酶基因反义RNA对淋巴瘤细胞Jurkat生长的影响.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2010 5 RNA Jurkat 1, 2 1 1, 2 1, 2 任玉珊 韩钰 蔡富强 王艳林 1 2 ( , 443002; , 443002) : 探讨鸟氨酸脱羧酶( ornithine decarboxylase, ODC)反义 RN 是否对人淋巴瘤细胞 Jurkat的 长具有抑制作用 含反义 RN 的真核表达载体 pcDN 3. 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, W estern blotting检测细胞中ODC蛋白表达水平,半定量 RTPCR检测细胞中ODC mRN 含量, 流式细胞术检测 细胞周期的变化, DN 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 RN 转染细 胞后, 引起 Jurkat细胞 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 RN 能抑制人 T淋巴瘤 Jurkat细胞的 长, 具有治疗人白血病的潜在应用价值 : 鸟氨酸脱羧酶 Jurkat细胞 反义 RN Effect of Antisense RNA of Ornithine Decarboxylase Gene on Jurkat Cell RenYushan1, 2 HanYu1 CaiFuqiang1, 2 W angYanlin1, 2 1 2 ( Institute of M olecularBiology, ThreeGorges University, Yichang 443002; MedicalCollege, Three Gorges University, Yichang 443002) A bstrac:t The study was designeol to explore the effects of antisenseRN of ornithine decarboxylase (ODC) on Jurkat cells. The eukaryotic expression plasmid for ODC antisense RN was constructed. The plasm id was then transfected into Jurkat cells with lipo fectam ine reagent. The cell linew ith ODC expression inhibition was selected by RTPCR andW estern blotting assay. Using this cell line as amodel, the effects ofODC antisenseRN on cellgrowth and cellshap(MTS assay) , cell cycle ( flow cytometry assay) and cell sensitive to DFMO (MTS andDN fragmentation assay)were determ ined. Results indicated that the plasm id pcDN 3. 1( + ) /Rodc for ODC antisense RN was transfected into Jurkat cells successfully and a Jurkat cell line ( J/o)