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安 威, 杜海军, 陈 莉
(首都医科大学细胞生物系, 100054 北京)
摘 要:
本文通过研究转染人肝刺激物质(hepatic stimulator substance, HSS)的肝癌细胞增殖状态,
进一步探讨了该基因的生物学功能。将人HSS基因导入BEL-7402肝癌细胞, 用Northern
和Southern杂交法证实该基因在靶细胞中有稳定表达。并通过测定细胞生长曲线, 细胞S期比例和细胞MAPK活性, 观察到转HSS基因BEL-7402细胞增殖发生了改变。实验结果提示,
HSS表达的肝癌细胞DNA合成增加, 增殖速度加快, 可能与MAPK激活有关。HSS基因表达可促进细胞增殖。
关键词:
肝刺激物质; 基因转染; 肝癌细胞增殖
学科分类号: Q485; R333.4
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AN Wei, DU Hai-jun, CHEN Li
(Department of Cell Biology, Capital University of Medical Sciences,
Beijing 100054)
Abstract:
A hepatotropic factor originally identified in weanling or regenerating
rat livers, known as hepatic stimulator substance (HSS), is characterized
by promoting markedly liver regeneration after partial hepatectomy.
Although HSS gene and the gene product have been described recently, the
cellular mechanism of HSS and its genetic function remain obscure. In our
previous studies, human HSS (hHSS) was demonstrated to promote the growth
of hepatoma cells and phosphorylate the mitogen-activated protein kinase (MAPK).
In this study, we reported the growth features of human hepatoma cells
response to hHSS gene transfection. The hHSS eukaryotic vector was
transfected to BEL-7402 hepatoma cells and the expression of hHSS was
analyzed with Northern and Southern blot. The results showed that the HSS
recombinant construct was functionally expressed in the target cells as
analyzed with Northern blot and reverse transcriptase polymerase chain
reaction (RT-PCR). The growth potential of HSS-transfected hepatoma cells
was markedly enhanced as compared with the non-transfected cells. The
S-phase of the hHSS-transfected cells increased by 61.5% than that of the
non-transfected cells as shown by flow cytometry. Moreover, MAPK
phosphorylation, one of the most important mitogenic indexes of
growth signal cascade, was profoundly activated at sites of Thr202/Tyr204
due to HSS gene transfer. Based on these results, it is concluded that the
introduction of HSS gene into hepatoma cells might be able to stimulate
directly the cellular growth in vitro, allowing a possibility of
reevaluation of HSS mechanism in intact cells.
Key words:
hepatic stimulatory substance; gene transfection; liver proliferation
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