4-氨基-2-三氟甲基苯基维甲酸酯通过PTEN/PI3K/Akt抑制YAC-1细胞增殖和诱导其分化  

Effects of 4-amino-2-trifluoromethyl-phenyl retinate on proliferation and differentiation of YAC-1 cells via PTEN /PI3K /Akt

作　　者：彭晓清[1] 陈飞虎[1] 葛金芳[1] 汪楠[1] 潘春晓[1] 雷静[1] 

机构地区：[1]安徽医科大学药学院,安徽合肥230032

出　　处：《中国药理学通报》2014年第1期60-65,共6页Chinese Pharmacological Bulletin

基　　金：国家重大新药创制专项(No 2009ZX09103386)

摘　　要：目的研究新型维甲酸衍生物4-氨基-2-三氟甲基苯基维甲酸酯（4-amino-2-trifluoromethyl—phenylretinate，ATPR）对鼠淋巴瘤YAC-1细胞增殖和分化的作用及其可能机制。方法不同浓度的ATPR作用鼠淋巴瘤YAC-1细胞后，MTT检测细胞生长抑制率；LDH法检测细胞毒性；瑞氏-吉姆萨染色法检测细胞形态；NBT检测细胞分化阳性率；RT—PCR和Westernblot法检测维甲酸受体mRNA和蛋白表达变化；Westernblot法检测FFEN／P13K／Akt和cyclinD的蛋白表达。结果ATPR（10^-5、10^-6、10^-7、10^-8、10^-9mol·L^-1）用药后72h明显抑制YAC-1细胞增殖，其抑制作用随浓度和时间增加而逐渐增强；随着药物浓度升高，ATPR对YAC-1细胞毒性增强；ATPR（10^-5～10^-9mol·L^-1）作用72h后可诱导YAC-1细胞形态呈现高分化趋势，且NBT阳性率升高；ATPR（10^-5mol·L^-1）体外用药72h可诱导RARα的mR—NA和蛋白表达升高，但RARβ和RARγ表达无明显变化；而且可诱导PTEN表达升高并下调p-Akt和cyclinD的蛋白表达。结论ATPR可明显抑制YAC-1细胞增殖并诱导其分化，其机制可能是与RARα结合后参与调节PTEN／P13K／Akt信号通路，从而抑制cyclinD的过表达。Aim To investigate the effects of 4-amino- 2-trifluoromethyl-phenyl retinate （ATPR） on the pro- liferation and differentiation of mouse lymphoma cancer YAC-1 cells and its possible mechanisms. Methods YAC-1 cells were cultured and treated with ATPR at different concentrations. The proliferation of YAC-1 cells was evaluated using 3- （ 4,5-Dimethyhhiazol-2- yl） -2, 5- diphenyltetrazolium bromide （MTT）, and the cell toxicity was detected by LDH （lactate dehydro- genase） assay. Morphological changes were observed via Wright-Giemsa staining. The positive cell ratio was assessed by NBT （Nitrotetrazolium Blue chloride）as- say. The mRNA and protein expressions of retinoic acid receptors （RARα, RARe, and RARe） were de- tected by reverse transcriptase-polymerase chain reac- tion （RT-PCR） and Western blot, respectively. The protein expressions of FFEN, Akt, p-Akt, cyclinD were detected by Western blot. Results The prolifer- ation of YAC-1 cells was inhibited by treatment with ATPR （10-5 - 10-9 mol ·L-l）, and the inhibition was increased as the concentration had increased. The best inhibitory effect reached the peak at 72 h. Cell toxicity was increased as the concentration had in- creased. YAC-1 cells treated with ATPR（ 10-5 - 10-9 mol · L-l）presented higher degree of mature and dif- ferentiation through Wright-Giemsa staining and NBT. The protein expressions of p-Akt, cyclinD were re- markably decreased （ P 〈 0. 05 ）, while the expression of PTEN and RARα were notably increased （P 〈 0. 05 ）, with no significant difference in the expressions of RARe, RARα and Akt among groups. Conclusion ATPR could significantly inhibit the proliferation and induce differentiation of mouse lymphoma YAC-1 cells, the mechanism of which might be involved in its ability in mediating the expression of retinoic acid receptors, and regulating the PTEN/PDK/Akt signaling pathway and cyclin D expression eventually.

关 键 词：4-氨基-2-三氟甲基苯基维甲酸酯 淋巴瘤 细胞增 殖 细胞分化 YAC-1细胞 维甲酸衍生物 PTEN P13K AKT 

分 类 号：R-332[医药卫生] R329.24