摘要
目的 观察核转录因子-κB抑制蛋白激酶（IKKβ）小干扰RNA（siRNA）对耐替莫唑胺（TMZ）的胶质瘤细胞株TR-U251和TR-LN229增殖的影响,探讨IKKβ干扰在耐药细胞TMZ化疗敏感性中的作用.方法 体外分步诱导法建立TMZ耐药细胞株TR-U251和TR-LN229,分别用25、50、100 μmol/L siRNA转染耐药细胞,逆转录-聚合酶链反应（RT-PCR）法鉴定IKKβ mRNA表达量；噻唑蓝（MTT）法测定50 μmol/L IKKβ siRNA干扰前后各组细胞TMZ半数抑制浓度（IC50）值并计算耐药指数；转染50 μmol/L IKKβ siRNA的耐药细胞分别联合或不联合IC20TMZ,MTT法检测细胞的增殖抑制率.结果 25 μmol/L IKKβ siRNA干扰效果最弱,50μmol/L和100 μmol/L效果较强；进行50 μmol/L IKKβ siRNA干扰24 h,TR-U251干扰前后对TMZ耐药指数分别为5.92±0.66和0.45±0.07；TR-LN229干扰前后分别为6.58 ±0.29和0.22±0.10;50 μmol/L IKKβ siRNA单独作用TR-U251和TR-LN229细胞48 h增殖抑制率分别为（17.47±4.11）％、（5.90±2.81）％,120μmol/LTMZ单独作用TR-U251 48 h细胞增殖抑制率为（7.87±3.29）％,60 μmol/L TMZ单独作用TR-LN229 48 h细胞增殖率为（24.67±5.37）％,而IKKβ干扰与TMZ联合作用48 h,TR-U251和TR-LN229细胞增殖抑制率分别为（66.33±9.18）％、（75.30±6.01）％.结论 IKKβ RNA干扰显著降低TR-U251与TR-LN229对TMZ的耐药指数及增殖率,增加TMZ化疗的敏感性.
Objective To observe the effects of nuclear factor-κB kinases β（IKKβ） small interfering RNA （siRNA） on the proliferation and apoptosis in temozolomide （TMZ）-resistant glioma cell lines TR-U251 and TR-LN229,and to explore the role of IKKβ siRNA in TMZ chemotherapy sensitivity.Methods TMZ resistant cell lines TR-U251 and TR-LN229 were constructed by stepwise exposure of parental cells to TMZ in vitro.The TMZ resistant cells were transfected by 25,50 and 100 μmol/L siRNA respectively and then reverse transcriptase-polymerase chain reaction （RT-PCR） was used to detect the IKKβ mRNA expression levels.The TMZ resistance index of TR-U251 and TR-LN229 cells was analyzed by methyl thiazol tetrazolium （MTT） with or without transfection of 50 μmol/L siRNA for 24 h.The resistant cells were divided into 50 μmol/L siRNA transfection for 48 h group and non-transfection group,respectively,combined with or without IC20 TMZ.The cell proliferation inhibition rate was analyzed by MTT.Results Both 50 and 100 μmol/L siRNA inhibited more significantly the IKKβ mRNA expression than 25 μmol/L siRNA.The TMZ resistance index of TR-U251 and TR-LN229 cells was respectively 5.92 ±0.66 and 6.58 ±0.29,however,after transfection by 50 μmoL/L siRNA for 24 h,the TMZ resistance index of TR-U251 and TR-LN229 cells was respectively 0.45 ± 0.07 and 0.22 ± 0.10.The proliferation inhibition rate for TR-U251 and TR-LN229 cells was respectively （17.47 ±4.11） %,（5.90 ±2.81） % in 50 μmol/L IKKβ siRNA alone,and that was （7.87 ± 3.29）% in 120 μmol/L TMZ alone for TR-U251 group,（24.67 ± 5.37） % in 60 μmol/L TMZ alone for TR-LN229 group,however,when combined with IKKβ siRNA for 48 h,TR-U251 and TR-LN229 cell proliferation inhibition rate was respectively （66.33 ± 9.18） %,（75.30 ± 6.01） %.Conclusion The IKKβ RNA interference significantly decreased the TMZ resistance index and the proliferation,thus increased the TMZ chemotherapy sensitivity in TR-U251 and TR-LN229 cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第11期2330-2332,共3页
Chinese Journal of Experimental Surgery
