重组腺病毒载体介导共刺激分子B7.1基因对儿童肝母细胞瘤体外淋巴细胞增值的影响

目的研究含hB7-1基因的重组腺病毒载体对儿童肝母细胞瘤体外淋巴细胞增值的影响。方法以腺病毒为载体,将hB7-1基因转入HepG2细胞,以FCM检测HepG2和HepG2/hB7-1细胞表面B7-1分子的表达情况。观察HepG2和HepG2/hB7-1细胞的生长曲线。用混合淋巴细胞培养法检测瘤苗对T淋巴细胞增殖的影响。结果HepG2细胞表面B7-1分子表达水平低,其阳性率为1.1%。而HepG2/hB7-1的B7-1分子表达阳性率达到18.5%,B7-1基因的表达水平提高16.8倍。HepG2和HepG2/BB-102的生长曲线相比,后者生长曲线上升减缓,生长速度减慢。B7-1分子的表达促进了淋巴细胞的增殖,较对照组有显著性差异(P<0.05)。结论1.HepG2表面B7-1分子低水平表达是其弱免疫原性的主要原因。2.含hB7-1基因的重组腺病毒载体转染HepG2可以获得B7-1分子高表达的细胞株。3.重组腺病毒载体的转染可以降低肝母细胞瘤细胞的恶性表征。4.转染含hB7-1基因的瘤苗可以促进T淋巴细胞的增殖。

Effect of the hepatoblastoma by transfection of adenovirus-mediated human B7-1 gene on proliferation of lymphocytes in vitro

Objective:To investigate the proliferation of lymphocytes after in vitro adenoviral transfection of hB7-1 gene into hepatoblastoma derived from children.Methods:After hB7-1 cells were transfected with B7-1 gene,the expression of B7-1 in HepG2 cells and HepG2 cells growth were determined by flow cytometric analysis.MTT Assay was also used to assess the proliferation of lymphocyte that was co-cultured with HepG2 cells and HepG2/hB7-1 cells.Results:Flow cytometry revealed that HepG2 cells have a very low expression of B7-1 on the cell surface(positive rate:1.1%).Conversely,the HepG2/B7-1 clone strongly expressed B7-1(positive rate:18.5%,increased 16.8 times compared to the former) on the cell surface.After transfection with B7-1 gene,the HepG2 cell proliferation slowed down.The results further revealed that HepG/B7-1 cells stimulated vigorous proliferation of lymphocytes.Conclusion:The low expression of B7-1 is the main reason for HepG2 cell to be poorly immunogenic.The transduction of B7-1 gene could enhance B7-1 expression in HepG cell,and at the same time inhibit the proliferation of HepG cell,and stimulate vigorous proliferation of lymphocytes in vitro.