摘要
目的探讨转铁蛋白受体(TFRC)在替莫唑胺抑制U87胶质瘤细胞增殖和侵袭中的作用。方法体外培养U87胶质瘤细胞,加入50μmol/L、100μmol/L和200μmol/L替莫唑胺作用;构建control siRNA、siTFRC转染U87细胞沉默TFRC表达,构建pcDNA3.1空载体、pcDNA3.1/TFRC转染U87细胞过表达TFRC;利用CCK-8方法检测U87细胞增殖;利用Transwell小室实验检测U87细胞侵袭能力;实时荧光定量PCR和免疫印迹法检查U87细胞TFRC mRNA和蛋白表达。结果与空白组相比,替莫唑胺处理后,U87细胞的增殖和侵袭能力显著下降(P<0.05),TFRC mRNA和蛋白表达水平显著降低(P<0.05),当替莫唑胺浓度为200μmol/L时,对U87细胞的抑制能力最强(P<0.05)。当利用pcDNA3.1/TFRC过表达U87细胞TFRC处理后,替莫唑胺对U87细胞增殖和侵袭的抑制能力显著下降(P<0.05);而当利用siTFRC沉默U87细胞TFRC表达后,替莫唑胺对U87细胞增殖和侵袭的抑制能力显著增强(P<0.05)。结论替莫唑胺可能通过抑制TFRC的表达而抑制U87胶质瘤细胞的增殖和侵袭。
Objective To investigate the role of transferrin receptor(TFRC)in the inhibition of proliferation and invasion of U87 glioma cells by temozolomide.Methods U87 glioma cells were cultured in vitro,and treated with 50μmol/L,100μmol/L and 200μmol/L temozolamine.Control siRNA and siTFRC transfected into U87 cells were used to silence TFRC expression,and pcDNA3.1 empty vector and pcDNA3.1 transfected into U87 cells were used to over-express TFRC expression.U87 cell proliferation was detected by CCK-8 method.U87 cell invasion ability was detected by Transwell chamber assay.TFRC mRNA and protein expression in U87 cells were detected by real-time fluorescent quantitative PCR and immunoblotting,respectively.Results Compared with the blank group(without treatment of temozolomide),the proliferation and invasion ability of U87 cells reduced significantly(P<0.05)and the expression of TFRC mRNA and protein decreased significantly(P<0.05)after temozolomide treatment,in a concentration-dependent manner.When the concentration of temozolomide was 200μmol/L,the inhibition ability of temozolomide to U87 cells was the strongest(P<0.05).Compared with the U87 cells treated with pcDNA3.1 empty vector,the expression level of TFRC significantly increased in the U87 cells treated with pcDNA3.1/TFRC(P<0.05).The inhibitory effect of temozolomide on cell proliferation and invasion was significantly reduced in U87 cells treated with pcDNA3.1/TFRC compared to those treated with pcDNA3.1 empty vector(P<0.05).Compared with the U87 cells treated with control siRNA,the expression level of TFRC significantly decreased in the U87 cells treated with siTFRC(P<0.05).The inhibitory effect of temozolomide on cell proliferation and invasion was significantly increased in U87 cells treated with siTFRC compared to those treated with control siRNA(P<0.05).Conclusion Temozolomide may inhibit the proliferation and invasion of U87 glioma cells by inhibiting the expression of TFRC.
作者
赵海康
潘力
张亮
冯小云
刘建荣
蒋小兵
高文文
马廉亭
ZHAO Hai-kang;PAN Li;ZHANG Liang;FENG Xiao-yun;LIU Jian-rong;JIANG Xiao-bing;GAO Wen-wen;MA Lianting(Department of Neurosurgery,General Hospital,Central Theater,PLA,Wuhan 430070,China;Department of Neurosurgery,The Second Affiliated Hospital,Xi'an Medical College,Xi'an 710038,China;Department of Neurosurgery,General Hospital,Tianjin Medical University,Tianjin 300052,China)
出处
《中国临床神经外科杂志》
2019年第11期685-689,共5页
Chinese Journal of Clinical Neurosurgery
