IκB激酶β小分子干扰RNA增敏人胶质瘤细胞替莫唑胺化疗的机制  

Mechanism of IkB kinase βtargeting small interfering RNA on enhanced temozolomide chemotherapy sensitivity in human glioma cells

作　　者：陈祎阳[1] 洪新雨[2] 王焕宇[1] 

机构地区：[1]天津市环湖医院神经外科天津市脑血管与神经变性重点实验室,300060 [2]吉林大学第一医院神经外科

出　　处：《中华实验外科杂志》2014年第4期779-781,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察IKB激酶B（IKKβ）小干扰RNA（siRNA）对耐替莫唑胺（TMZ）的胶质瘤细胞凋亡的影响，探讨IKKβ siRNA增敏耐药细胞TMZ化疗的机制。方法构建人胶质瘤耐药细胞株TR-U251和TR-LN229细胞，随机分为对照组、50μmol/LIKβsiRNA转染组、低浓度IC20 TMZ治疗组以及50μmol/LIKKβsiRNA＋IC2。TMZ联合治疗组，流式细胞术检测细胞凋亡率，Western blot法检测B细胞淋巴瘤/白血病-2（bcl-2）、半胱氨酰天冬氨酸特异性蛋白酶-3（Caspase-3）表达，以及干扰前后核转录因子．KB（NF-KB）p65、0^6-甲基鸟嘌呤-DNA-甲基转移酶（MGMT）、Survivin蛋白表达水平。结果TR-U251细胞凋亡率分别为：对照组（2．78±1．67）％、干扰组（11．92±3．06）％、120txmol/LTMZ治疗组（8．18±2．24）％、联合治疗组（35．95±6．07）％；TR-LN229细胞凋亡率分别为：对照组（1．824-0．86）％、干扰组（9．784-4．13）％、60μmol／LTMZ组（8．65±3．64）％、联合治疗组（28．30±4．97）％，两种细胞联合治疗组凋亡率较对照组和单独用药组均显著增加（P〈0．01），联合治疗组bcl-2水平均显著降低，Caspase-3水平均增加；IKK[3RNA干扰后NF-KB、MGMT及Survivin蛋白表达抑制。结论IKK[3RNA干扰通过显著降低TR-U251与TR-LN229中NF-KB、MGMT及Survivin蛋白表达，增高TMZ化疗的凋亡率。Objective To observe the effects of IKB kinase β （IKKβ） small interfering RNA （siRNA） on the apoptosis of temozolomide （TMZ）-resistant glioma cell lines TR-U251 and TR-LN229, and to explore the mechanism of IKKβ siRNA in TMZ chemotherapy sensitivity. Methods TMZ resistant cell lines TR-U251 and TR-LN229 were constructed, then the TMZ resistant cells were randomly devided into control group, 50 p, mol/L IKKI3 siRNA group, IC20 TMZ therapy group and 50 μmol/L IKK[3 siRNA ＋ IC20 TMZ combination therapy group. The apoptosis rate was examined by flow cytometry. The protein expres sion levels of B cell lymphoma/leukemia-2 （ bel-2 ） , Cysteinyl aspartate-specific protease-3 （ Caspase-3 ） and nuclear factor-KB （NF-KB） 1365, O6-methylguanine-DNA methytransferase （ MGMT）, and Survivin with or without IKK[3 siRNA transfection were detected by Western blotting. Results The apoptosis rate in TR-U251 cells in control group, 50 μmol/L IKK[3 siRNA group, TMZ therapy group and combination ther apy group was （2.78 ± 1.67）% , （11.92 ±3.06）% , （8.18 ±2.24）% and and （35.95 ±6.07）%, and that inTR-LN229 cells was （1.82±0.86）%, （9.78 ±4.13）%, （8.65±3.64）% and （28.30± 4. 97 ） % , respectively. The apoptosis rate in the combination therapy group was obviously high than other groups in two cell linces （ P 〈 0. 01 ）. Additionally, the protein bcl-2 expression levels were suppressed, and those of Caspase-3 were increased in the conbination therapy group; the levels of NF-KB, MGMT and Survivin were all suppressed with IKKβ siRNA transfeetion in two cell linces. Conclusion The IKKβ RNA interference significantly increased the apoptosis rate in TMZ chemotherapy by inhibiting the expres sion levels of NF-KB, MGMT and Survivinin in TR-U251 and TR-LN229 cell lines.

关 键 词：胶质瘤 化疗耐药 替莫唑胺 IKB激酶β 

分 类 号：R739.4[医药卫生—肿瘤]