中文摘要：

目的分析替莫唑胺（TMZ）对胶质瘤U251细胞及其干细胞livin、MRP基因表达及细胞周期的影响。方法分离培养U251细胞及其干细胞后，不同浓度的TMZ（0、25、50、100、200、400μmol／L）干预72h，CCK-8检测细胞增殖变化趋势，流式细胞术（FCM）检测细胞周期变化，RT-PCR技术检测livin、MRP1、MRP3mRNA表达。结果本研究成功从U251细胞中分离出U251干细胞；与未干预组相比，25μmol／L的TMZ干预U251细胞及其干细胞后其增殖率分别为0．952±0．011（t=-8．219，P=0．001）、0．937±0．029（t=-3．822，P=0．019）和100μmol／L的TMZ干预U251细胞及其干细胞后其增殖率分别为0．750±0．018、0．887±0．039（t=5．480，P=0．005）；正常情况下，livin基因在U251细胞及其干细胞中的表达量分别为（0．411±0．025）×10^-3、（0．571±0．040）×10^-3（t=-3．348，P=0．004），MRP1基因的表达量分别为（0．295±0．018）×^-3、（0．481±0．034）×10^-3（t=-8．439，P=0．001），MRP3基因的表达量分别为（1．128±0．117）×10^-3、（0．963±0．059）×10^-3（t=2．176，P=0．095）；200μmol／L的TMZ干预U251细胞及其干细胞后livinmRNA表达量在分另0为（0．020±0．002）×10^-3（t=27．123，P=0）、（0．066±0．007）×10^-3（t=21．345，P=0）；200μmol／L干预后MRP1 mRNA表达量为（0．556±0．041）×10^-3（t=-10．214，P=0．001）、（0．6094-0．044）×10^-3（t=-3．98，P=0．016）；200μmol／L干预MRP3mRNA表达量为（1．397±0．192）×10^-3（t=-2．073，P=0．107）、（1．496±0．302）×10^-3（t=-2．993，P=0．040）；100μmol／L干预后细胞周期U251细胞G2／M期百分比为11．353±0．516、39．1±1．2（t=-37．1，P=0）和U251干细胞S期百分比为21．1±1．0、30．2±1．7（t=-8．07，P=0．001）。结论替莫唑胺能有效降低U251及其干细胞中livin基因的表达，并使细胞分裂周期停滞，降低细胞增殖率，促?

英文摘要：

Objective To analyze the effects of temozolomide （TMZ） on glioma U251 cells and their stem cell livin, MRP gene expression and cell cycle. Methods After U251 cells and their stem cells being isolated and cultured, the different concentrations of TMZ （0, 25, 50, 100, 200, and 400μmol/L） were used to intervene for 72 h. CCK-8 was used to detect the change trend of cell proliferation. Flow eytometry （FCM） was used to detect the changes of cell cycle. RT-PCR technique was used to detect the expression levels of livin, MRP1, and MRP3 mRNA. Results Our study successfully isolated stem cells U251from U251 cells. Compared with a non-intervention group, after 25μmol/L TMZ intervention of 15251 cells and their stem cells, the proliferation rate were 0. 952 ＋ 0. 011 （ t = - 8. 219, P = 0. 001 ） and 0. 937±0. 029 （t = - 3. 822, P = 0. 019） respectively, and after 100 μmol/L TMZ intervention of U251 ceils and their stem cells, the proliferation rate were 0. 750 ±0. 018 and 0. 887±0. 039 （t =5. 480, P = 0. 005 ） respectively. Under the normal circumstances, the expression level of livin gene in U251 cells and their stem cells were 0.411 ±0.025 × 10-3and 0.571 ±0.040 × 10-3（t = -3.348, P =0.004） respectively; the expression level of MRP1 gene were 0. 295±0. 018 × 10-3and 0. 481 ±0. 034 × 10-3 （t = -8. 439, P = 0. 001 ） respectively; the expression level of MRP3 gene were 1. 128 ± 0. 117× 10-3and 0. 963±0. 059× 10-3 （t = 2. 176, P = 0. 095） respectively. After 200μmol/L TMZ intervention of U251 cells and their stem cells, the expression levels of livin mRNA were 0. 020 × 0. 002× 10-3 （ t = 27. 123, P = 0） and 0. 066 ± 0. 007 × 10-3 （t = 21. 345, P = 0 ） respectively. After the intervention of 200 p.mol/L, the expression levels of MRP1 mRNA were 0. 556 ＋ 0. 041 × 10-3 （t = - 10. 214, P = 0. 001 ） and 0. 609 ＋ 0. 044× 10-3（ t = - 3.98, P = 0. 016 ） ; after the intervention of 200μmol/L, the expression levels of MRP3 mRNA were 1. 397 ± 0. 192 × 10