论文部分内容阅读
为了探索新城疫病毒血凝素神经氨酸酶(HN)基因对人肺癌细胞SPC-A1的作用及机制,将含有HN基因的重组质粒pVHN经脂质体介导转染人肺癌细胞SPC-A1,通过MTT方法检测细胞活力;采用吖啶橙/溴化乙锭(AO/EB)染色分析肿瘤细胞凋亡;罗丹明123和DCFA染色测定线粒体跨膜电位(ΔΨm)和活性氧水平变化;流式细胞仪分析MHC-Ⅰ分子表达;底物染色反应检测caspase-3活性.结果重组质粒pVHN转染人肺癌细胞SPC-A148h后,细胞活力明显降低;AO/EB染色可见明显的细胞凋亡形态学变化;与空质粒对照相比,线粒体ΔΨm下降(P<0·01),活性氧水平升高(P<0·05);细胞表面MHC-Ⅰ分子表达上调(P>0·05);caspase-3活性增强(P<0·01).以上结果提示,新城疫病毒HN基因能够上调SPC-A1细胞表面MHC-Ⅰ分子表达,并通过上调ROS水平,下调线粒体ΔΨm,进而激活caspase-3,最终诱导人肺癌细胞凋亡.
In order to explore the effect and mechanism of the hemagglutinin neuraminidase (HN) gene on human lung cancer cell line SPC-A1, the recombinant plasmid pVHN containing HN gene was transfected into human lung cancer cell line SPC-A1 Cell viability was detected by MTT assay. Apoptosis of tumor cells was analyzed by AO / EB staining. Mitochondrial transmembrane potential (ΔΨm) and reactive oxygen species (ROS) levels were measured by rhodamine 123 and DCFA staining. The expression of MHC-Ⅰ was detected by cytometry, and the activity of caspase-3 was detected by substrate staining.Results The viability of transfected human lung cancer cell SPC-A148h was significantly decreased after the recombinant plasmid pVHN was transfected.Apoptotic morphology was observed by AO / EB staining (P <0.01), reactive oxygen species (P <0.05), and up-regulated expression of MHC-I on the cell surface (P> 0.05). (P <0.01) .The above results suggest that HN gene of Newcastle disease virus can up-regulate the expression of MHC-Ⅰ on the surface of SPC-A1 cells and downregulate the mitochondrial ΔΨm by upregulating the level of ROS, and then activate caspase-3 , Eventually inducing apoptosis in human lung cancer cells.