新城疫病毒FMW株体外溶瘤作用及其机制

Mechanisms of in vitro anti-tumor effects of a Newcastle disease virus

摘要:

摘要：【目的】筛选出能高效抑制多种人肿瘤细胞生长增殖的新城疫（New Castle disease virus, NDV）毒株，为进一步构建重组高效靶向溶瘤毒株奠定基础。【方法】以体外噻唑蓝法测定NDV对A549、 SMMC7721等肿瘤细胞及人胚干细胞L-O2、人胚肾细胞HEK293等的生长抑制率，空斑试验确定病毒滴度及感染复数。利用形态学观察、 Hoechst荧光染色、流式细胞术及免疫印迹等分析了NDV-FMW诱导肿瘤细胞凋亡的细胞生物学变化及其机制。【结果】从近50株NDV中筛选出NDV-FMW，以

Abstract:

Abstract: [Objective] To obtain Newcastle disease virus (NDV) strains with high in vitro anti-tumor effect for construction of recombinant NDV for clinical therapy. [Methods] We used MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphemyltetra-zolium Bromide] assay to examine the anti-tumor effect on A549 and SMMC7721 cells infected by nearly 50 NDV strains. Several assays were used to analyze the apoptosis induced by NDV infection. These assays included:(1) Morphological analysis; (2) Hoechst? fluorescence dye testing; (3) Flow cytometric analysis, and (4) Western blot. [Results] We obtained an NDV FMW strain that inhibited cell growth up to 60% at 48 h postinfection with an MOI (multiplex of infection) of 20. NDV- FMW could elicit apoptosis of infected tumor cells in a dose- and time- dependent manner. We observed the infected A549 and SMMC7721 cells with condensed and fragmented chromatin at 48 h postinfection. Apoptosis peak and hypodiploid cells were revealed by proidum iodide (PI) staining and cell cycle was blocked and arrested in G0/G1 phase in tested cells. Furthermore, annexin-ⅴ/PI staining showed that the apoptotic rates in SMMC7721 cells were 2.1% ,18.5%, 23.8% and 30.4% after treated with 0, 0.2, 2 and 20 MOI NDV-FMW for 48 h respectively. To elucidate the apoptosis pathways induced by NDV-FMW, we detected the expression of active caspase-3 and cleavages of poly(ADP-ribose) polymerase (PARP). We demonstrated that caspase-3 in A549 cells was activated early at 16 h postinfection and PARP was cleaved subsequently. [Conclusion] NDV-FMW had strong in vitro anti-tumor effect on A549 and SMMC7721 cells. Apoptosis of tumor cells induced by NDV-FMW via caspase-3 activation and NDV-FMW could be a potential cancer virotherapy agent.