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目的:探讨倍半萜烯内酯(sesquiterpene Iactones,SLs)类化合物对不同环氧合酶-2(cyclooxygenase-2,COX-2)表达水平的人鼻咽癌(nasopharyngeal carcinoma,NPC)细胞中癌干细胞(cancer stem cells,CSCs)转归的影响。方法:分别采用实时定量PCR(qRT-PCR)和Western blot检测人鼻咽癌高分化CNE1L和低分化CNE2L细胞株中COX-2 mRNA和蛋白本底表达水平。给予不同剂量SLs活性成分小白菊内酯(parthenolide,PN)处理2种细胞株后,再分别采用噻唑蓝(MTT)法测定细胞增殖活性;qRT-PCR检测细胞中干性相关基因COX-2、ABCG2、BMI1、OCT4 mRNA水平;流式细胞术分析干性侧群(side population,SP)细胞的相对含量;平板克隆集落形成实验检测干性细胞的自我更新和分化增殖能力。结果:CNE1L和CNE2L细胞中COX-2 mRNA和蛋白本底水平显著不同,在CNE1L细胞中明显高于CNE2L细胞(P<0.05);与对照组比较,PN处理后,细胞增殖抑制率随PN浓度(10～100μmol/L)增加呈剂量依赖性增高(P<0.05),细胞中COX-2 mRNA在CNE1L中升高、在CNE2L中降低,干性相关基因ABCG2、BMI1 mRNA在CNE1L细胞中升高、而OCT4 mRNA在两株细胞中均降低(P均<0.05);SP细胞抑制率亦随PN浓度(2.5～40μmol/L作用24 h)增加呈剂量依赖性增高(P<0.05);而细胞的集落形成能力随PN浓度(0.25～1.5μmol/L作用72 h)增加显著降低(P<0.05);上述结果在CNE1L和CNE2L细胞株间差异均有统计学意义(P<0.05),在CNE1L细胞中变化作用更强。结论:PN对不同NPC细胞株中的干性细胞功能具有抑制作用且存在明显差别,可能与不同细胞间COX-2表达的差异性有关。
Objective: To investigate the effect of sesquiterpene Iactones (SLs) on the expression of cyclooxygenase-2 (COX-2) in human nasopharyngeal carcinoma (NPC) Impact of stem cell (CSCs) outcome. Methods: Real-time quantitative PCR (qRT-PCR) and Western blot were used to detect the expression of COX-2 mRNA and protein in human nasopharyngeal carcinoma differentiated CNE1L and poorly differentiated CNE2L cell lines respectively. After treated with parthenolide (PN) at different dosages, two cell lines were treated with MTT assay. The expressions of COX-2, ABCG2, BMI1 and OCT4 mRNA levels were detected by flow cytometry. The relative content of side population (SP) cells was analyzed by flow cytometry. The ability of self-renewal and differentiation and proliferation of stem cells were detected by plate clone colony formation assay. Results: The mRNA and protein levels of COX-2 in CNE1L and CNE2L cells were significantly different from those in CNE1L cells and CNE2L cells (P <0.05). Compared with the control group, the inhibition rate of PNX (10 ~ 100μmol / L) increased in a dose-dependent manner (P <0.05). COX-2 mRNA increased in CNE1L cells, decreased in CNE2L cells and ABCG2 mRNA and BMI1 mRNA increased in CNE1L cells , While OCT4 mRNA decreased in both cell lines (all P <0.05). The inhibition rate of SP cells also increased in a dose-dependent manner with the increase of PN concentration (2.5-40 micromol / L for 24 h) (P <0.05) (P <0.05). The above results were significantly different between CNE1L and CNE2L cell lines (P <0.05). In CNE1L The role of changes in cells stronger. CONCLUSION: PN inhibits the function of stem cells in different NPC cell lines and has significant difference, which may be related to the different expression of COX-2 in different cell lines.