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目的:探讨Th1类细胞因子IFN-γ和Th2类细胞因子IL-4对MCF-7细胞的成瘤、黏附能力的调节作用及其相关机制。方法:常规体外培养人乳腺癌细胞系MCF-7。以前期工作中筛选出的以有效剂量rh IFN-γ(100 ng/ml)或rh IL-4(10ng/ml)或细胞因子稀释液分别处理细胞96 h后,应用半定量RT-PCR法、Western blotting技术、双层软琼脂集落形成实验、细胞体外黏附实验等观察IFN-γ和IL-4对细胞VEGF表达、致瘤性和黏附侵袭特性的影响,并对其相关信号通路机制进行分析。结果:IFN-γ或IL-4可分别抑制或促进人乳腺癌细胞MCF-7的成瘤和黏附能力。分子机制研究表明,与对照组相比,rh IFN-γ可抑制MCF-7细胞VEGF、MMP-9的基因转录和蛋白表达水平,抑制Raf/MEK/ERK信号通路;rh IL-4可促进MCF-7细胞的VEGF、MMP-2和MMP-9的基因转录和蛋白表达水平,促进PI3K/AKT、Raf/MEK/ERK信号通路激活。结论:肿瘤微环境中,IFN-γ和IL-4可分别抑制和促进人乳腺癌细胞系MCF-7的VEGF表达和体外成瘤、黏附能力,其机制可能与PI3K/AKT、Raf/MEK/ERK信号通路相关。
Objective: To investigate the regulatory effect of Th1 cytokine IFN-γ and Th2 cytokine IL-4 on tumorigenesis and adhesion of MCF-7 cells and its related mechanisms. Methods: Human breast cancer cell line MCF-7 was routinely cultured in vitro. The cells were treated with rh IFN-γ (100 ng / ml) or rhIL-4 (10 ng / ml) or cytokines dilution for 96 h, and then semi-quantitative RT- The effects of IFN-γ and IL-4 on the expression of VEGF, tumorigenicity and adhesion and invasion of cells were observed by Western blotting, double soft agar colony formation assay and in vitro cell adhesion assay. The related signaling pathways were also analyzed. Results: IFN-γ or IL-4 could inhibit or promote tumorigenesis and adhesion of human breast cancer cell MCF-7 respectively. Compared with the control group, rh IFN-γ inhibited the gene transcription and protein expression of VEGF, MMP-9 and inhibited the Raf / MEK / ERK signaling pathway in MCF-7 cells. Rh IL-4 promoted MCF- -7 cells, and promote the gene transcription and protein expression of VEGF, MMP-2 and MMP-9 and promote the activation of PI3K / AKT and Raf / MEK / ERK signaling pathways. Conclusions: IFN-γ and IL-4 in tumor microenvironment can inhibit and promote VEGF expression and in vitro tumorigenesis and adhesion of human breast cancer cell line MCF-7, respectively. The mechanism may be related to PI3K / AKT, Raf / MEK / ERK signaling pathway related.