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目的:研究全反式维甲酸(all trans retinoic acid,ATRA)对人胃腺癌细胞(SGC-7901)细胞周期阻滞的诱导作用并探讨其机制。方法:采用四甲基偶氮唑盐(MTT)方法检测ATRA对SGC-7901增殖的抑制;流式细胞术检测细胞周期,Westernblot方法检测不同浓度ATRA处理后的SGC-7901细胞中Akt、p-Ak(tSer473)、p-Ak(tThr308)、p-GSK-3β(Ser9)和cyclinD1的表达情况。结果:10-9～10-5mol/L的ATRA作用SGC-7901细胞48h,能显著抑制细胞增殖,其抑制率分别为10.2%±0.5%、15.3%±0.5%、17.0%±0.7%、28.4%±1.0%和36.9%±0.7%;G1期细胞比率随着ATRA浓度的增加而增加,呈明显的G1期阻滞;Westernblot检测显示ATRA对细胞中Akt蛋白的表达没有明显影响,两种p-Akt蛋白的表达显著下调,ATRA显著降低细胞中cyclinD1和p-GSK-3β的表达。结论:ATRA可能通过抑制磷酸化Akt蛋白表达而减少p-GSK-3β的表达,从而减少cyclinD1的表达量,进而诱导SGC-7901细胞发生G1期阻滞。
Objective: To investigate the induction of all-trans retinoic acid (ATRA) on the cell cycle arrest of human gastric adenocarcinoma cell line SGC-7901 and its mechanism. Methods: The inhibition of ATRA on the proliferation of SGC-7901 cells was detected by MTT assay. The cell cycle was detected by flow cytometry. The expressions of Akt, p-Akt in SGC-7901 cells treated with different concentrations of ATRA were detected by Western blot. Akt (tSer473), p-Ak (tThr308), p-GSK-3β (Ser9) and cyclinD1. Results: SGC-7901 cells treated with 10-9 ~ 10-5mol / L ATRA for 48h significantly inhibited cell proliferation with the inhibitory rates of 10.2% ± 0.5%, 15.3% ± 0.5%, 17.0% ± 0.7%, 28.4 % ± 1.0% and 36.9% ± 0.7% respectively. The percentage of cells in G1 phase increased with the increase of ATRA concentration and showed obvious G1 arrest. Western blot analysis showed that ATRA had no significant effect on the expression of Akt protein in cells. Akt protein expression was significantly down-regulated, ATRA significantly reduced the expression of cyclinD1 and p-GSK-3β. CONCLUSION: ATRA may reduce the expression of p-GSK-3β, and thus reduce the expression of cyclinD1, and thus induce G1 arrest in SGC-7901 cells by inhibiting the expression of phosphorylated Akt protein.