摘要目的 观察多重基因转染对神经母细胞瘤SH-SY5Y细胞增殖、凋亡及免疫原性的影响.方法 以空病毒质粒(SH/Ad组)、携带肿瘤抑制基因人野生型p53(wt-p53)的腺病毒质粒(SH/p53组)及携带wt-D53、免疫相关基因B7-1和粒-巨噬细胞集落刺激因子(CM-CSF)基因的腺病毒质粒(SH/BB-102组)转染SH-SY5Y细胞,并以未转染的肿瘤细胞为空白对照(SH组).分别采用免疫印迹法、流式细胞术和酶联免疫吸附法检测p53蛋白、B-1分子和GM-CSF的表达.观察转染前后细胞生长及凋亡情况.用混合淋巴细胞培养法检测瘤苗对T细胞增殖和细胞因子分泌的诱导作用.结果 腺病毒转染SH-SY5Y细胞后,目的基因得到了高效表达.与SH和SH/Ad组相比,SH/p63和SH/BB-102组细胞生长速度减慢,凋亡增加(P<0.05).SH/BB-102组与其他组比较,能够促进T细胞增殖,增加γ干扰素和白细胞介素-2的分泌(P<0.05).结论 wt-p53、E7-1和GM-CSF基因联合转染可以抑制SH-SY5Y细胞的生长、诱导细胞凋亡,并且可以增强肿瘤细胞的免疫原性.

abstractsObjective To observe the effeet of cotransduction of wt-p53,B7-1 and GM-CSF genes mediated by recombinant adenovirus on cell growth, apoptosis and immunogenicity of neuroblastoma cell line SH-SY5Y, Methods We transfected SH-SYSY cells separately with recombinant adenovirus, recombinant adenovirus mediating human wild type p53(wt-p53) gene and BB-102.We separately named them with SH/Ad,SH/p53 and SH/BB-102 group.The protein expression of p53, B7-1 and GM-CSF were measured by Western blot, FCM and enzyme linked immunosorbent assay. Cell number was counted and growth curves were drawed. Cell apoptosis was tested by FCM. Lymphocyte proliferation and cytokine secretory was evaluated by mixed lymphocyte culturing. Results After transection with BB- 102, p53, B7-1, and GM-CSF, high-efficiency expression of target genes were found. Growth of the cells was inhibited, apoptosis was induced in the SH/p53 and SH/BB-102 groups. Proliferation of lymphocytes was stimulated and cytokine increased visorously in the SH/BB-102 group. Condusions After transfection with BB-102, growth of the SH-SY5Y cells was inhibited,apoptosis was induced,and immunogenicity was greatly enhanced.