医学论文-新城疫病毒体外诱导S180细胞凋亡的实验研究.docVIP

医学论文-新城疫病毒体外诱导S180细胞凋亡的实验研究 ???????????????????? 作者：蒋丽娜 孙黎 程建贞 罗强 翟登高? 【摘要】? 目的：探讨新城疫病毒(Newcastle disease virus，NDV)体外对S180肉瘤细胞（简称S180）生长有无抑制和诱导细胞凋亡作用。方法:取培养至对数生长期的S180细胞浓度为5×104/mL和1×106/mL分别接种于96孔板和50mL培养瓶中，加入不同浓度的NDV(1∶20、1∶10、1∶5)作用24h，分别用MTT法检测NDV对S180瘤细胞的生长抑制作用,荧光显微镜和琼脂糖凝胶电泳检测细胞凋亡。结果：经上述不同浓度的NDV作用24h，对S180细胞生长的抑制率依次为10%、26%、39%，各实验组吸光度值（A490）与对照组比较差异均有统计学意义(P0.05)。荧光显微镜观察实验组培养的S180瘤细胞，出现细胞核固缩及凋亡小体，琼脂糖凝胶电泳见DNA梯形条带。结论：NDV对体外培养S180细胞增殖有明显抑制作用，可诱导S180细胞凋亡。 【关键词】? 新城疫病毒；细胞凋亡;电泳/琼脂凝胶 ??? 【ABSTRACT】? Objective:To investigate whether the Newcastle disease virus can inhibit proliferation of S180 cells and/or induce apoptosis of the cells in vitro.Methods:S180 cells（5×104/mL）were cultured at midexponential phase and planted in 96 well plates with 100μL each and in 50mL culture bottles,1.5mL each.The cells were treated for 24 hours with different concentrations of Newcastle disease virus.Growth inhibition rate was measured by MTT method.Apoptosis was detected by the following ways:fluorescence microscopy with Annexin VFITC staining was used for observing the change of cell morphology，agarose electrophoresis was used to detect the DNA changes and observe subdiploid.Results:Newcastle disease virus could inhibit proliferation of s180 cells.The inhibition rates for S180 cells treated with Newcastle disease virus were 10%,26%，39% under the Newcastle disease virus concentrations of 1∶20、1∶10、1∶5 respectively,with a significant difference to that of the control（t=3.606,P0.05；t= 4.327，P0.05；t=4.321,P0.05）.Cell contraction and apoptotic bodies were observed under fluorescence microscope.DNA fragment was shown through agarose electrophoresis.Conclusion:Newcastle disease virus can inhibits proliferation and induces cell apoptosis in S180 cells in vitro. ??? 【KEY WORDS】? Newcastle disease virus,Apoptosis,Electrophoresis/Ager Gel ??? 肿瘤的发生不仅与细胞的过度增殖有关,而且与细胞凋亡受抑制从而使已转化的细胞生存期限延长有关。细胞凋亡具可诱导性,因此,诱导肿瘤细胞凋亡成为肿瘤治疗的一大热点[1]。近年来，关于NDV感染诱导肿瘤细胞凋亡报道比较多，而对于S180细胞凋亡的诱导作用报道较少。本实验研究了NDV体外感染S180细胞的