Jiang Hong-li, Xue Wu-jun, Liu Hua, Chen Wei
Hemodialysis Center, The First Clinical Medical College, Xi'an Jiaotong University, Xi'an 710061, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 Jan;22(1):47-50.
To investigate the inhibitory effects of dumb-bell decoy oligodeoxynucleotides(ODNs) targeting NF-kappaB on the transcriptional activity of NF-kappaB binding to its cis-elements, growth of multiple myeloma cells 8266 and the expression of IL-6 in 8266 cells.
The inhibitory effects of dumbbell decoy ODNs targeting NF-kappaB on the transcriptional activity of NF-kappaB was detected by electrophoretic mobility shift assay(EMSA). 8266 cells were randomly divided into 8266 cells of passage (control), dumb-bell decoy ODNs treatment, and cationic liposome treatment groups. Dumb-bell decoy ODNs of 2, 4 and 8 mg/L were cultured respectively into 8266 cells via cationic liposome vector. Then, the transfected 8266 cells and their supernatant were collected at 8, 12 and 18 h. IL-6 expression was measured by ELISA. Effect of decoy ODNs on the growth of 8266 cells stimulated with IL-6 was measured by MTT Colorimetry assay.
Dumbbell decoy ODNs effectively inhibited the binding of NF-kappaB to its cis-elements in vitro. Different doses of decoy ODNs (2, 4 and 8 mg/L) had different inhibitory effects on IL-6 expression in 8266 cells. After decoy ODNs had been transfected into 8266 cells stimulated by IL-6 via cationic liposome vector, both the growth of 8266 cells and activity of IL-6 were inhibited.
Dumb-bell decoy ODNs can effectively inhibit the growth of 8266 cells and decrease the expression of IL-6 in myeloma cells through the inhibiting NF-kappaB activity.
研究靶向核因子-κB（NF-κB）的哑铃型诱饵寡脱氧核苷酸（ODNs）对NF-κB与其顺式作用元件结合的转录活性、多发性骨髓瘤细胞8266生长及8266细胞中白细胞介素-6（IL-6）表达的抑制作用。
采用电泳迁移率变动分析（EMSA）检测靶向NF-κB的哑铃型诱饵ODNs对NF-κB转录活性的抑制作用。将8266细胞随机分为传代8266细胞（对照）、哑铃型诱饵ODNs处理组和阳离子脂质体处理组。分别将2、4和8 mg/L的哑铃型诱饵ODNs通过阳离子脂质体载体培养至8266细胞中。然后，在8、12和18小时收集转染后的8266细胞及其上清液。采用酶联免疫吸附测定（ELISA）法检测IL-6表达。采用噻唑蓝（MTT）比色法检测诱饵ODNs对IL-6刺激的8266细胞生长的影响。
哑铃型诱饵ODNs在体外能有效抑制NF-κB与其顺式作用元件的结合。不同剂量的诱饵ODNs（2、4和8 mg/L）对8266细胞中IL-6表达有不同的抑制作用。通过阳离子脂质体载体将诱饵ODNs转染至经IL-6刺激的8266细胞后，8266细胞的生长及IL-6活性均受到抑制。
哑铃型诱饵ODNs可通过抑制NF-κB活性有效抑制8266细胞生长并降低骨髓瘤细胞中IL-6的表达。
