摘要
目的探讨塞来昔布对辐射后鼻咽癌(NPC)残癌细胞的干细胞表型标志物表达的影响。方法通过连续7次2 Gy 60钴(60Co)γ射线照射CNE-1细胞,建立辐射后CNE-1残癌细胞株E-R7。通过CCK-8实验,筛选塞来昔布对CNE-1残癌细胞的最佳药物浓度;采用实时定量聚合酶链式反应、免疫印迹检测塞来昔布对CNE-1残癌细胞干细胞表型标志物表达的影响。结果 CCK-8实验结果表明,E-R7细胞的塞来昔布半抑制浓度为131μM。按照塞来昔布半抑制浓度,给予E-R7细胞塞来昔布处理。实时定量聚合酶链式反应测定结果显示,塞来昔布处理后,E-R7细胞中干细胞表型标志物性别决定区Y框蛋白2(SOX2)、八聚体结合转录因子4(OCT4)、Kruppel样因子4(KLF4)的mRNA表达水平较未处理的E-R7细胞明显降低,差异有统计学意义(P<0.05)。免疫印迹检测结果显示,塞来昔布处理后,E-R7细胞中干细胞表型标志物SOX2、OCT4、KLF4的蛋白表达水平较未处理的E-R7细胞明显降低,差异有统计学意义(P<0.05)。结论塞来昔布可抑制经2 Gy ^60Coγ射线连续照射的CNE-1细胞获得的残癌细胞干细胞表型标志物SOX2、OCT4、KLF4的表达。
Objective To investigate the effect of celecoxib on the expression of stem cell phenotypic markers in nasopharyngeal carcinoma(NPC)residual cancer cells.Methods CNE-1 cells were irradiated with 2 Gy 60Co γ ray for 7 times in a row to establish the residual CNE-1 cancer cell strain E-R7 after radiation.CCK-8 assay was used to screen the best concentration of celecoxib against CNE-1 residual cancer cells.Real-time quantitative polymerase chain reaction and Western blot were used to detect the effect of celecoxib on the expression of phenotypic markers of CNE-1 residual cancer stem cells.Results CCK-8 results showed that the celecoxib semi-inhibitory concentration of E-R7 cells was 131 μM.E-R7 cells were treated with celecoxib at celecoxib semi-inhibitory concentration.Real-time quantitative polymerase chain reaction assay showed that after celecoxib treatment,the mRNA expression levels of stem cell phenotype markers SOX2,OCT4 and KLF4 in E-R7 cells were significantly lower than those in untreated E-R7 cells,with statistically significant differences(P<0.05).Western blot test results showed that after celecoxib treatment,the protein expression levels of the stem cell phenotype markers SOX2,OCT4 and KLF4 in E-R7 cells were significantly lower than those in untreated E-R7 cells(P<0.05).Conclusion Celecoxib inhibits the expression of SOX2,OCT4 and KLF4,phenotypic markers of residual cancer stem cells obtained by CNE-1 cells continuously irradiated with 2 Gy ^60Co γ ray.
作者
李菁菁
姜曈
石梅
LI Jing-jing;JIANG Tong;SHI Mei(Department of Radiotherapy,The First Hospital Affiliated to the Fourth Military University,Xi'an 710032,China)
出处
《创伤与急危重病医学》
2019年第3期140-144,共5页
Trauma and Critical Care Medicine
基金
国家自然科学基金(81372421)
关键词
辐射
鼻咽癌
干细胞表型
塞来昔布
Radiation
Nasopharyngeal carcinoma
Stem cell phenotype
Celecoxib
