Differentiation Induction Effect Of4-Amino-2-Trifluoromethyl-Phenyl Retinate On Lymphoma Cancer Cell And Its Possible Mechanisms

As one of the differentiation inducing agents, all-trans retinoic acid (ATRA) hasgenerally been applied to cure acute promyelocytic leukemia successfully, with the goodfunction of inducing tumor cancer cells to a higher degree of mature. However, theside-effects of ATRA including retinoic acid syndrome and RA resistance limited itsfurther application in the clinical. we reformed the polar group at the end of the carbonchain and modified the cyclohexene ring with all-trans retinoic acid(ATRA) as the leadcompound, synthesized a series of new retinoic acid derivatives which had beendepurated and identified. After screening of the derivatives with structure-activity,4-amino-2-trifluoromethyl-phenyl retinate (ATPR) has powerful activity in inducingcell differentiation. This research select lymphoma cells including mouse lymphatictumor YAC-1cell and human MCL cell Jeko-1as objects, to reaserch the inductiondifferentiation activities of ATPR on lymphoma cancer cells and then explore itspotential mechanisms.Objective: Reaserch the induction differentiation effect of ATPR on lymphoma cancercells and explore its potential mechanisms. Contents:1. To observe the effects of induction differentiation of ATPR in YAC-1and Jeko-1cells.2. To observe the effects of ATPR on RARs in YAC-1and Jeko-1cells.3. To explore the role of PTEN/PI3K/Akt signaling in the process of inductiondifferentiation of ATPR in YAC-1and Jeko-1cells.Methods:1. Using MTT assay to detect the proliferation of YAC-1and Jeko-1cells.Morphologic changes were observed via Wright-Giemsa staining. The positive cell ratiowas assessed by NBT assay. Using FCS to observe the expression of an exclusive cellsurface antigen CD38and the distribution of cell cycle on Jeko-1cells.2. The mRNA and protein expressions of retinoic acid receptors (RARα, RARβ, andRARγ) were detected by RT-PCR and western blotting, respectively.3. The protein expressions of PTEN, pAkt, cyclinD were tested by western blotting inYAC-1cells after treatment; RNA interference technique used to silence PTEN genein Jeko-1cell, then using western blotting to investigated the protein expression of Akt,pAkt, cyclinD.Results:1. ATPR could effectively induce YAC-1and Jeko-1cells indifferentiaion: Theproliferation of YAC-1and Jeko-1cells was inhibited by treatment with ATPR (10-5~10-9mol·L-1) in a dose-and time-dependent manner; YAC-1and Jeko-1cells treatedwith ATPR presented higher degree of mature and differentiation through Wright-Giemsa staining and NBT. The percentage of cells in G0/G1phase wasincreased, and the expression level of the maturation specific cell surface marker CD38increased in Jeko-1cells.2. ATPR combines with RARs to exert induction differentiation effects: the mRNAand protein expression of RARα in YAC-1cells both raised. Likely, the mRNA andprotein expression of RARα and RARr in Jeko-1cells both increased.3. ATPR modulates PTEN/PI3K/Akt signaling to exert induction differentiationeffects: the protein expressions of pAkt, cyclinD were remarkably decreased, while theexpression of PTEN was notably increased in YAC-1cells. After PTEN gene wassilenced selectively for6h, Jeko-1cells were putted in ATPR for72h, western blottingresults diaplayed that the protein expression of p-Akt and cyclinD was increased,compared with the ATPR group without silencing (P<0.01).Conclusion:1. The differentiation of YAC-1and Jeko-1cells could be induced by ATPR.2. ATPR combines with RARs and modulates PTEN/PI3K/Akt signaling pathway toexert induction differentiation function.