Moriya Tetsuhiro, Kita Kazuko, Sugaya Shigeru, Wano Chieko, Suzuki Nobuo
Department of Environmental Biochemistry, Graduate School of Medicine, Chiba University, Chiba City, Chiba, Japan.
Biol Sci Space. 2002 Mar;16(1):12-7. doi: 10.2187/bss.16.12.
A major issue in radiation and space biology is whether gene expression levels are altered in cells exposed to gravity-changing stress. In the present study, genes up- or down-regulated in radiation-sensitive human RSa cells cultured under gravity-changing conditions, were identified using a PCR-based mRNA differential display method. Exposure of cells to gravity-changing stress was performed by free-fall with a drop-shaft facility or by an airplane-conducted parabolic flight. Among the candidates for gravity-changing stress-responsive genes obtained by the differential display analysis, the lactate dehydrogenase A gene (LDH-A) was confirmed by Northern blotting analysis to exhibit increased expression levels. The gravity-changing stress consisted of a combination of microgravity and hypergravity. However, exposure of the cells to hypergravity produced by centrifuge only slightly affected the LDH-A mRNA expression. Thus, LDH-A was found to be a candidate for the genes which play a role in the cellular response to gravity-changing stress, and mainly to microgravity.
辐射与空间生物学中的一个主要问题是,暴露于重力变化应激下的细胞中基因表达水平是否会发生改变。在本研究中,使用基于聚合酶链反应(PCR)的mRNA差异显示方法,鉴定了在重力变化条件下培养的辐射敏感型人RSa细胞中上调或下调的基因。通过使用落塔设施进行自由落体或通过飞机进行抛物线飞行,使细胞暴露于重力变化应激。在通过差异显示分析获得的重力变化应激反应基因候选物中,通过Northern印迹分析证实乳酸脱氢酶A基因(LDH-A)表达水平升高。重力变化应激由微重力和超重力组合而成。然而,仅通过离心机使细胞暴露于超重力对LDH-A mRNA表达的影响很小。因此,发现LDH-A是在细胞对重力变化应激(主要是微重力)的反应中起作用的基因候选物。