Rémy-Heintz N, Duperray C, Bali J P
Laboratoire de Biochimie des Membranes, INSERM CJF92-07, Faculté de Pharmacie, Montpellier.
C R Seances Soc Biol Fil. 1993;187(5):678-88.
The growth of the human gastric cancer cell line HGT-1 is regulated by a gastrin-like peptide through an autocrine process. In order to analyse the mechanism of action of this peptide, a study at different steps of the cell cycle was considered; so, a blocking of this cell line by thymidine or hydroxy-urea was studied by cytofluorimetry. In normal growth conditions in 10% FCS medium, 57% of the cells were in G0/G1 phase and 31% in S phase. A treatment with 2 mM hydroxy-urea followed by 4 hours in 10% FCS medium led to 85% of the cells in S phase. By successive treatments with thymidine and hydroxy-urea followed by 1 hour in 10% FCS medium, 2 peaks of S phase corresponding to 86% of the cells were observed; after 24 hours, cells were distributed as found for the unconfluent cell line, whatever the treatment. On the other hand, the thymidine kinase activity of unconfluent cells which was relatively elevated as compared to other cell lines (278 mU/mg protein), was increased by synchronisation with hydroxyurea followed by 1 hour in 10% SVF medium (338 mU/mg protein); after 8 hours in 10% FCS medium, this activity decreased at the value observed for cells treated with thymidine followed by 1 hour in 10% FCS medium (214 mU/mg protein). In conclusion, a synchronisation either by thymidine or by hydroxy-urea, led to a blocking of the HGT-1 cell line at different steps of the cell cycle, leading to a better knowledge of its autocrine growth regulation.
人胃癌细胞系HGT - 1的生长受一种胃泌素样肽通过自分泌过程调控。为分析该肽的作用机制,考虑了在细胞周期不同阶段进行研究;因此,通过细胞荧光测定法研究了用胸苷或羟基脲对该细胞系的阻断作用。在含10%胎牛血清(FCS)培养基的正常生长条件下,57%的细胞处于G0/G1期,31%处于S期。用2 mM羟基脲处理后在含10% FCS培养基中培养4小时,导致85%的细胞处于S期。通过连续用胸苷和羟基脲处理,随后在含10% FCS培养基中培养1小时,观察到对应86%细胞的2个S期峰;24小时后,无论处理方式如何,细胞分布与未汇合细胞系相同。另一方面,与其他细胞系相比未汇合细胞的胸苷激酶活性相对较高(278 mU/mg蛋白),经羟基脲同步化处理后在含10%血清替代物(SVF)培养基中培养1小时,该活性增加(338 mU/mg蛋白);在含10% FCS培养基中培养8小时后,该活性降至用胸苷处理后在含10% FCS培养基中培养1小时的细胞所观察到的值(214 mU/mg蛋白)。总之,用胸苷或羟基脲进行同步化处理,导致HGT - 1细胞系在细胞周期的不同阶段被阻断,从而更深入了解其自分泌生长调控。