全反式维甲酸和亚硒酸钠对HL-60细胞Rb基因的RNA表达及其产物磷酸化的影响  

Effects of all-trans retinoic acid and sodium selenite on RNAof retinoblastoma gene (Rb) of human myeloid leukemia cells(HL 60) and phosphorylation of its gene product

作　　者：刘新光[1,2] 于树玉[1,2] 

机构地区：[1]广东医学院医用生化研究所 [2]中国医学科学院肿瘤研究所生化室

出　　处：《癌症》1998年第6期435-437,464,共4页Chinese Journal of Cancer

摘　　要：目的：观察全反式维甲酸（ＲＡ）和亚硒酸钠对ＨＬ６０的Ｒｂ基因的ＲＮＡ及其产物磷酸化的联合效应。方法：以ＨＬ６０细胞为实验对象，利用狭线杂交、Ｗｅｓｔｅｒｎｂｌｏｔｉｎｇ和流式细胞仪进行研究。结果：狭线杂交实验表明ＲＡ和硒单独或联合都对Ｒｂ基因的ＲＮＡ无明显影响。Ｗｅｓｔｅｒｎｂｌｏｔｉｎｇ结果显示：ＲＡ和硒可影响该细胞ＲＢ蛋白磷酸化与去磷酸化状态。在第４天０．１μｍｏｌ／ＬＲＡ可使去磷酸化ＲＢ蛋白增加４７％；５．８μｍｏｌ／ＬＮａ２ＳｅＯ３可使磷酸化ＲＢ蛋白减少４９％，对去磷酸化ＲＢ蛋白仅轻度增加。联合后去磷酸化ＲＢ蛋白明显增加，较对照组增加了１２７％，明显高于两单独组。实验中ＲＢ蛋白磷酸化状态与细胞分化和细胞周期关系与文献报道一致。结论：ＲＡ和硒联用对ＨＬ６０细胞的ＲＢ蛋白产物去磷酸化作用比单独作用更强。Objective: Effects of all trans retinoic acid (RA) and sodium selenite (Na2SeO3) on RNA of retinoblastoma gene (Rb) of HL 60 cell line and phosphorylation of its gene product was studied. Methods: Slot hybridization, westerm blotting and flow cytometric analysis were undertaken. Results: Slot hybridization of RNA demonstrated that neither single or combined use of RA and sodium selenite had no significant effects on Rb gene. Western blotting analysis showed that RA and sodium selenite influenced phosphorylation of retinoblasoma protein in HL-60 cells. 4 days after treatment of 0.1 μmol/L RA, cells expressed hypophosphorylation RB protein majorly, increased 47% that of control group. In the case of 5 8 μmol/L Na2SeO3, the amount of the phosphorylated RB protein decreased some 49% than that of control while the hypophosphorylated RB protein increased a little. Expression of hypophosphorylaed RB protein was much higher than that of the phosphorylated RB protein after combined treatment, the amount of the hypophosphorylated RB protein incerased 127% than that of control. In the experiment, the relationships of RB protein phosphorylation state and the results of the cell differentiation and cell cycle were according to the previous reports. Conclusion: Combined effects of retinoic acid (0 1 μmol/L RA) and sodium selenite (5.8 μmol/L Na2SeO3) on dephosphorylation of RB protein were much stronger than either of two chemicals during HL 60 cell proliferation and differentiation.

关 键 词：全反式维甲酸 亚硒酸钠 HL60细胞 白血病 

分 类 号：R733.710.5[医药卫生—肿瘤] R730.53[医药卫生—临床医学]