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

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

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。

Objective：To investigate the proliferation of lymphocytes after in vitro adenoviral transfection of hB7 - 1 gene into hepgatoblastoma 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.