中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
11期
2330-2332
,共3页
胶质瘤%化疗耐药%替莫唑胺%核转录因子-κB抑制蛋白激酶
膠質瘤%化療耐藥%替莫唑胺%覈轉錄因子-κB抑製蛋白激酶
효질류%화료내약%체막서알%핵전록인자-κB억제단백격매
Glioma%Chemotherapy-resistance%Temozolomide%Nuclear factor-κB kinases β
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.
目的 觀察覈轉錄因子-κB抑製蛋白激酶(IKKβ)小榦擾RNA(siRNA)對耐替莫唑胺(TMZ)的膠質瘤細胞株TR-U251和TR-LN229增殖的影響,探討IKKβ榦擾在耐藥細胞TMZ化療敏感性中的作用.方法 體外分步誘導法建立TMZ耐藥細胞株TR-U251和TR-LN229,分彆用25、50、100 μmol/L siRNA轉染耐藥細胞,逆轉錄-聚閤酶鏈反應(RT-PCR)法鑒定IKKβ mRNA錶達量；噻唑藍(MTT)法測定50 μmol/L IKKβ siRNA榦擾前後各組細胞TMZ半數抑製濃度(IC50)值併計算耐藥指數；轉染50 μmol/L IKKβ siRNA的耐藥細胞分彆聯閤或不聯閤IC20TMZ,MTT法檢測細胞的增殖抑製率.結果 25 μmol/L IKKβ siRNA榦擾效果最弱,50μmol/L和100 μmol/L效果較彊；進行50 μmol/L IKKβ siRNA榦擾24 h,TR-U251榦擾前後對TMZ耐藥指數分彆為5.92±0.66和0.45±0.07；TR-LN229榦擾前後分彆為6.58 ±0.29和0.22±0.10;50 μmol/L IKKβ siRNA單獨作用TR-U251和TR-LN229細胞48 h增殖抑製率分彆為(17.47±4.11)％、(5.90±2.81)％,120μmol/LTMZ單獨作用TR-U251 48 h細胞增殖抑製率為(7.87±3.29)％,60 μmol/L TMZ單獨作用TR-LN229 48 h細胞增殖率為(24.67±5.37)％,而IKKβ榦擾與TMZ聯閤作用48 h,TR-U251和TR-LN229細胞增殖抑製率分彆為(66.33±9.18)％、(75.30±6.01)％.結論 IKKβ RNA榦擾顯著降低TR-U251與TR-LN229對TMZ的耐藥指數及增殖率,增加TMZ化療的敏感性.
목적 관찰핵전록인자-κB억제단백격매(IKKβ)소간우RNA(siRNA)대내체막서알(TMZ)적효질류세포주TR-U251화TR-LN229증식적영향,탐토IKKβ간우재내약세포TMZ화료민감성중적작용.방법 체외분보유도법건립TMZ내약세포주TR-U251화TR-LN229,분별용25、50、100 μmol/L siRNA전염내약세포,역전록-취합매련반응(RT-PCR)법감정IKKβ mRNA표체량；새서람(MTT)법측정50 μmol/L IKKβ siRNA간우전후각조세포TMZ반수억제농도(IC50)치병계산내약지수；전염50 μmol/L IKKβ siRNA적내약세포분별연합혹불연합IC20TMZ,MTT법검측세포적증식억제솔.결과 25 μmol/L IKKβ siRNA간우효과최약,50μmol/L화100 μmol/L효과교강；진행50 μmol/L IKKβ siRNA간우24 h,TR-U251간우전후대TMZ내약지수분별위5.92±0.66화0.45±0.07；TR-LN229간우전후분별위6.58 ±0.29화0.22±0.10;50 μmol/L IKKβ siRNA단독작용TR-U251화TR-LN229세포48 h증식억제솔분별위(17.47±4.11)％、(5.90±2.81)％,120μmol/LTMZ단독작용TR-U251 48 h세포증식억제솔위(7.87±3.29)％,60 μmol/L TMZ단독작용TR-LN229 48 h세포증식솔위(24.67±5.37)％,이IKKβ간우여TMZ연합작용48 h,TR-U251화TR-LN229세포증식억제솔분별위(66.33±9.18)％、(75.30±6.01)％.결론 IKKβ RNA간우현저강저TR-U251여TR-LN229대TMZ적내약지수급증식솔,증가TMZ화료적민감성.
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.
