Golczewski J A, Frenkel G D
Dept. of Biological Sciences, Rutgers University, Newark, NJ 07102.
Biol Trace Elem Res. 1989 Apr-May;20(1-2):115-26. doi: 10.1007/BF02919104.
The incorporation of radioactive selenium into cellular proteins and the effect of selenite on proliferation were examined in human (HeLa, HT-29, and IMR-90) and mouse (3T3 and CMT-93) cell lines. Proteins incorporating selenium were detected by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Major polypeptide subunits at 60, 23, 21, 19, and 16 kD were detected in the two tumorigenic and one normal human cell lines. The 23 kD polypeptide migrated to the same position on the gel as the major subunit of human erythrocyte glutathione peroxidase. In the mouse cells, the 60 kD polypeptide was almost entirely absent; four other major selenoproteins were detected, with molecular weights similar to those in the human cells. In both mouse and human cells, the same pattern of selenoproteins was observed irrespective of whether the cells were grown in medium containing 10% fetal bovine serum or in defined medium supplemented with 0.1 or 1 microM selenite, or with 1% serum. The effect of selenite on proliferation of HeLa, HT-29, and CMT-93 cells in medium supplemented with 10% fetal bovine serum and in serum-free medium was examined. At concentrations up to about 1 microM, selenite stimulated proliferation of the human cells slightly in serum-free medium but not in serum-supplemented medium. At concentrations of about 5 microM and higher selenite significantly inhibited proliferation of all cells in both types of media. In CMT-93 cells, this inhibition was greater in serum-free medium, but there were no significant differences in this regard in the human cells. These results demonstrate that selenium is stably incorporated into several polypeptides in human and mouse cells, that there are no significant differences in this regard among several cell lines, and slight differences between human and mouse cells. They further confirm that selenium can have a slight stimulatory effect on cell growth, and a much larger inhibitory effect, depending on its concentration.
在人(HeLa、HT - 29和IMR - 90)及小鼠(3T3和CMT - 93)细胞系中检测了放射性硒掺入细胞蛋白质的情况以及亚硒酸盐对细胞增殖的影响。通过一维十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳检测掺入硒的蛋白质。在两种致瘤性人细胞系和一种正常人细胞系中检测到了60、23、21、19和16 kD的主要多肽亚基。23 kD的多肽在凝胶上迁移到与人红细胞谷胱甘肽过氧化物酶主要亚基相同的位置。在小鼠细胞中,几乎完全没有60 kD的多肽;检测到了另外四种主要的硒蛋白,其分子量与人细胞中的相似。在小鼠和人细胞中,无论细胞是在含10%胎牛血清的培养基中生长,还是在添加了0.1或1 microM亚硒酸盐或1%血清的限定培养基中生长,都观察到相同的硒蛋白模式。检测了亚硒酸盐对在含10%胎牛血清的培养基和无血清培养基中HeLa、HT - 29和CMT - 93细胞增殖的影响。在浓度高达约1 microM时,亚硒酸盐在无血清培养基中对人细胞的增殖有轻微刺激作用,但在含血清培养基中则无此作用。在浓度约为5 microM及更高时,亚硒酸盐显著抑制两种培养基中所有细胞的增殖。在CMT - 93细胞中,这种抑制在无血清培养基中更大,但在人细胞中在这方面没有显著差异。这些结果表明,硒稳定地掺入人和小鼠细胞中的几种多肽中,在几种细胞系之间在这方面没有显著差异,而人和小鼠细胞之间存在轻微差异。它们进一步证实,硒根据其浓度可对细胞生长产生轻微的刺激作用和更大的抑制作用。