中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
4期
779-781
,共3页
胶质瘤%化疗耐药%替莫唑胺%IκB激酶β
膠質瘤%化療耐藥%替莫唑胺%IκB激酶β
효질류%화료내약%체막서알%IκB격매β
Glioma%Chemotherapy-resistance%Temozolomide%IκB kinase β
目的 观察IκB激酶β(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IKKβsiRNA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、50 μmol/L IKKβ siRNA转染组、低浓度IC20 TMZ治疗组以及50 μmol/L IKKβ siRNA+ IC20 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-κB(NF-κB) p65、O6-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)％、120 μmol/L TMZ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(1.82±0.86)％、干扰组(9.78±4.13)％、60 μmol/L TMZ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;IKKβ RNA干扰后NF-κB、MGMT及Survivin蛋白表达抑制.结论 IKKβ RNA干扰通过显著降低TR-U251与TR-LN229中NF-κB、MGMT及Survivin蛋白表达,增高TMZ化疗的凋亡率.
目的 觀察IκB激酶β(IKKβ)小榦擾RNA(siRNA)對耐替莫唑胺(TMZ)的膠質瘤細胞凋亡的影響,探討IKKβsiRNA增敏耐藥細胞TMZ化療的機製.方法 構建人膠質瘤耐藥細胞株TR-U251和TR-LN229細胞,隨機分為對照組、50 μmol/L IKKβ siRNA轉染組、低濃度IC20 TMZ治療組以及50 μmol/L IKKβ siRNA+ IC20 TMZ聯閤治療組,流式細胞術檢測細胞凋亡率,Western blot法檢測B細胞淋巴瘤/白血病-2(bcl-2)、半胱氨酰天鼕氨痠特異性蛋白酶-3(Caspase-3)錶達,以及榦擾前後覈轉錄因子-κB(NF-κB) p65、O6-甲基鳥嘌呤-DNA-甲基轉移酶(MGMT)、Survivin蛋白錶達水平.結果 TR-U251細胞凋亡率分彆為:對照組(2.78±1.67)％、榦擾組(11.92±3.06)％、120 μmol/L TMZ治療組(8.18±2.24)％、聯閤治療組(35.95±6.07)％;TR-LN229細胞凋亡率分彆為:對照組(1.82±0.86)％、榦擾組(9.78±4.13)％、60 μmol/L TMZ組(8.65±3.64)％、聯閤治療組(28.30±4.97)％,兩種細胞聯閤治療組凋亡率較對照組和單獨用藥組均顯著增加(P＜0.01),聯閤治療組bcl-2水平均顯著降低,Caspase-3水平均增加;IKKβ RNA榦擾後NF-κB、MGMT及Survivin蛋白錶達抑製.結論 IKKβ RNA榦擾通過顯著降低TR-U251與TR-LN229中NF-κB、MGMT及Survivin蛋白錶達,增高TMZ化療的凋亡率.
목적 관찰IκB격매β(IKKβ)소간우RNA(siRNA)대내체막서알(TMZ)적효질류세포조망적영향,탐토IKKβsiRNA증민내약세포TMZ화료적궤제.방법 구건인효질류내약세포주TR-U251화TR-LN229세포,수궤분위대조조、50 μmol/L IKKβ siRNA전염조、저농도IC20 TMZ치료조이급50 μmol/L IKKβ siRNA+ IC20 TMZ연합치료조,류식세포술검측세포조망솔,Western blot법검측B세포림파류/백혈병-2(bcl-2)、반광안선천동안산특이성단백매-3(Caspase-3)표체,이급간우전후핵전록인자-κB(NF-κB) p65、O6-갑기조표령-DNA-갑기전이매(MGMT)、Survivin단백표체수평.결과 TR-U251세포조망솔분별위:대조조(2.78±1.67)％、간우조(11.92±3.06)％、120 μmol/L TMZ치료조(8.18±2.24)％、연합치료조(35.95±6.07)％;TR-LN229세포조망솔분별위:대조조(1.82±0.86)％、간우조(9.78±4.13)％、60 μmol/L TMZ조(8.65±3.64)％、연합치료조(28.30±4.97)％,량충세포연합치료조조망솔교대조조화단독용약조균현저증가(P＜0.01),연합치료조bcl-2수평균현저강저,Caspase-3수평균증가;IKKβ RNA간우후NF-κB、MGMT급Survivin단백표체억제.결론 IKKβ RNA간우통과현저강저TR-U251여TR-LN229중NF-κB、MGMT급Survivin단백표체,증고TMZ화료적조망솔.
Objective To observe the effects of IκB 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 μmol/L IKKβ siRNA group,IC20 TMZ therapy group and 50 μmol/L IKKβ siRNA + IC20 TMZ combination therapy group.The apoptosis rate was examined by flow cytometry.The protein expression levels of B cell lymphoma/leukemia-2 (bcl-2),Cysteinyl aspartate-specific protease-3 (Caspase-3) and nuclear factor-κB (NF-κB) p65,O6-methylguanine-DNA methytransferase (MGMT),and Survivin with or without IKKβ siRNA transfection were detected by Western blotting.Results The apoptosis rate in TR-U251 cells in control group,50 μmol/L IKKβ siRNA group,TMZ therapy group and combination therapy group was (2.78±1.67)％,(11.92±3.06)％,(8.18±2.24)％ and and (35.95±6.07)％,and that in TR-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-κB,MGMT and Survivin were all suppressed with IKKβ siRNA transfection in two cell linces.Conclusion The IKKβ RNA interference significantly increased the apoptosis rate in TMZ chemotherapy by inhibiting the expression levels of NF-κB,MGMT and Survivinin in TR-U251 and TR-LN229 cell lines.
