替莫唑胺对Livin基因过表达的胶质瘤TJ905细胞及其干细胞增殖活性的影响  

Effect of temozolomide on cell proliferation in TJ905 glioma stem cells with Livin over-expression

作　　者：李根华[1] 冯嵩[1] 韩光魁[1] 马文渊 刘臣 孔令胜[1] 靳峰[1] Li Genhua;Feng Song;Han Guangkui;Ma Wenyuan;Liu Chen;Kong Lingsheng;Jin Feng(Department of Neurosurgery,Affiliated Hospital of Jining Medical University&Shandong Provincial Key Laboratory of Stem Cells and Neuro-oncology,Jining 272029,China;Clinical Medical College,Jining Medical University,Jining 272067,China)

机构地区：[1]济宁医学院附属医院神经外科暨山东省干细胞与神经肿瘤学重点实验室,济宁272029 [2]济宁医学院临床医学院,济宁272067

出　　处：《中华脑科疾病与康复杂志（电子版）》2020年第2期99-105,共7页Chinese Journal of Brain Diseases and Rehabilitation（Electronic Edition）

基　　金：山东省医药卫生科技发展计划项目(2016WS0180);山东省自然科学基金资助项目(ZR2016HL34);山东省高等学校科技计划项目(J16LL05);济宁医学院教师科研扶持基金(JYFC2018FKJ139)。

摘　　要：目的分析替莫唑胺(TMZ)干预Livin基因过表达的胶质瘤TJ905细胞及其干细胞模型后对细胞增殖活性的影响,以及对胶质瘤TJ905细胞和TJ905干细胞中Livin、Caspase-3/7的作用机制。方法使用免疫磁珠方法从胶质瘤TJ905细胞系中分选出CD133阳性胶质瘤干细胞,采用慢病毒过表达技术建立Livin过表达的细胞模型,使用不同浓度(0、25、50、100、200、400μmol/L)的TMZ干预细胞模型48 h,采用CCK-8、流式细胞术、反转录PCR检测药物干预后细胞增殖活性、细胞周期及Livin、Caspase-3/7基因的表达量。结果(1)胶质瘤TJ905细胞中Livin基因表达量明显高于同源TJ905干细胞,差异有统计学意义(P<0.05);(2)TMZ能抑制胶质瘤TJ905细胞及其干细胞的增殖活性,降低增殖速度,并且随药物浓度的增加抑制作用逐渐增强;(3)TMZ能抑制Livin基因的表达,诱导Caspase-3/7的表达,并且随药物浓度的增加抑制作用逐渐增强,差异有统计学意义(P<0.05);(4)Livin基因转染后胶质瘤干细胞细胞周期G0/G1增加明显,而TJ905细胞周期则是G2/M期少量增加;TMZ干预细胞模型后,TJ905干细胞过表达组在S、G2/M停滞,对照组则是在S期停滞;TJ905细胞过表达组在S期停滞,对照组G2/M则是低浓度诱导停滞,随药物浓度增加其诱导作用减弱,差异均有统计学意义(P<0.05)。结论TMZ可以通过抑制Livin基因的表达、诱导Caspase-3/7的表达、调控细胞周期变化来降低细胞增殖活性,促进细胞凋亡。Objective To analyze the effect of cell proliferation after temozolomide(TMZ)intervention in TJ905 cells and stem cells with Livin gene overexpression.To explore the mechanism for TMZ to Livin and Caspase-3/7 in glioma TJ905 cells and stem cells.Methods CD133 positive TJ905 glioma cells were separated by immunomagnetic beads to isolate glioma stem cells,to establish the overexpression cell models of Livin gene with lentivirus transfection technology.Livin over-expression cell models were intervened by different concentration of TMZ for 48 h.Cell counting kit-8 assay,flow cytometry,reverse transcription-PCR were used to analyze cell cycle to investigate cell proliferation and to detect the expression of Livin and Caspase-3/7 respectively,following treatment with various concentrations of TMZ(0,25,50,100,200 and 400μmol/L)for 48 h.Results(1)The expression of Livin gene in glioma TJ905 cells was significantly higher than that of TJ905 stem cells(P<0.05);(2)TMZ could inhibit the proliferation of glioma TJ905 cells and stem cells,and the inhibitory effect increased with the increase of drug concentration;(3)TMZ could inhibit the expression of Livin gene and induce the expression of Caspase-3/7.With the increase of drug concentration,the inhibitory effect gradually increased,and the difference was statistically significant(P<0.05);(4)After Livin gene transfection,the cell cycle G0/G1 of TJ905 stem cellsincreased significantly,while TJ905 cell cycle increased slightly in G2/M phase.after TMZ intervention,TJ905 stem cell overexpression group stopped in S and G2/M phase,while that in control group was in S phase.TJ905 cellsoverexpression group stopped in S phase in the overexpression group,while G2/M in the control group was induced by low concentration,the induction effect was weakened with the increase of drug concentration,and the differences were statistically significant(—<0.05).Conclusion TMZ can inhibit the expression of Livin gene,induce the expression of Caspase-3/7,and regulate the changes of cell cycle to re

关 键 词：LIVIN Caspase-3/7 TJ905干细胞 替莫唑胺 细胞增殖 

分 类 号：R739.41[医药卫生—肿瘤]