Abstract
背景与目的肺癌是我国发病率和死亡率最高的恶性肿瘤,其中非小细胞肺癌(non-small cell lung cancer, NSCLC)约占肺癌的80%。表皮生长因子受体酪氨酸激酶抑制剂(epidermal growth factor receptor-tyrosine kinase inhibitor, EGFR-TKI)靶向治疗已成为NSCLC临床治疗的主要手段,然而不可避免的耐药性出现极大限制了EGFR-TKI的治疗效果。叉头框蛋白C1(forkhead box protein C1, FOXC1)是叉头框蛋白家族重要成员,在NSCLC异常表达并参与调控NSCLC进展。本研究旨在探讨干扰FOXC1对NSCLC吉非替尼耐药的影响及可能机制。方法应用Western blot、免疫组化方法检测FOXC1在NSCLC吉非替尼耐药细胞和组织中的表达情况; 应用FOXC1 shRNA转染HCC827/GR细胞,筛选稳定干扰FOXC1的HCC827/GR细胞,采用新型四氮唑盐比色法(Methyl-thiazolyldiphenyl-sulfophenyl-tetrazolium bromide assay, MTS)法、流式细胞术及微球体形成实验检测细胞增殖、细胞凋亡及细胞自我更新能力; 采用实时荧光定量PCR(quantitative real-time PCR, qRT-PCR)、Western blot检测干性标志物SOX2、Nanog、OCT4和CD133的表达水平; 运用流式细胞术检测CD133的表达情况; 免疫组化检测耐药组织中SOX2和CD133的表达情况; 基于癌症基因组图谱(The Cancer Genome Atlas, TCGA)数据库的肺腺癌数据集,分析FOXC1、SOX2和CD133表达的相关性。结果FOXC1在NSCLC吉非替尼耐药细胞HCC827/GR的表达水平显著高于HCC827敏感细胞(P < 0.05),免疫组化结果显示FOXC1在NSCLC吉非替尼耐药组织中高表达。与对照组相比,稳定干扰FOXC1的HCC827/GR细胞对吉非替尼的半数抑制浓度(50% inhibitory concentration, IC50)值显著降低(P < 0.01),细胞增殖能力降低、凋亡率升高(P < 0.05)。干扰FOXC1能够抑制SOX2、CD133的表达,并减弱HCC827/GR耐药细胞的微球体形成能力。免疫组化结果显示,吉非替尼耐药组织中SOX2和CD133表达显著高于敏感组织(P < 0.01)。相关性分析结果显示,FOXC1、SOX2和CD133表达两两呈正相关(P < 0.05)。结论FOXC1参与NSCLC吉非替尼耐药,其机制可能与FOXC1调控肿瘤干细胞(cancer stem cells, CSCs)特性有关。
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.