Effect Of4-Amino-2-Trifluoromethyl-Phenyl Retinate On The Differentiation And The Expression Of Retinoid Acid Receptors And Retinoid X Receptors Of Human Solid Tumor Cells

Objective: To investigate the proliferation inhibition and differentiationinduction activities of4-amino-2-trifluoromethyl-phenyl retinate（ATPR） inhuman esophageal cancer cells ECA-109, human gastric cancer cellsMGC80-3, human pancreatic cancer cells PANC-1, human cervical cancercells HeLa, human neuroblastoma cells SH-SY5Y.Explore the mechanisms ofATPR partly.Methods:1..Cell proliferation was assessed by MTT assay before and after thetreatment with ATPR(10^-4、10^-5、10^-6、10^-7、10^-8、10^-9mol/L) in vitro. Morphologicchanges were observed after Giemsa staining using inverted phase contrastmicroscope. ALP and LDH were measured by spectrophotometer for enzymeactivity; SCC, CA199and CA125were measured by ELISA; NSE wasmeasured by Immunocytochemistry; The cell cycle was analyzed by flowcytometry.2. Set up the control group （adding volume DMEM）, the ancholo group（containing0.05%ethanol）, positive control group of ATRA (10^-5mol/L), and ATPR of the experimental group (10^-5mol/L), The expression of RARα、RARβ、RARγ、RXRα、RXRβ and RXRγ mRNA treatment with ATPR for72h weredetected by RT-PCR and compare the differences in MGC80-3and SH-SY5Y.3. The expression of RARα、RARβ、RARγ、RXRα、RXRβ and RXRγ mRNAafter the treatment with ATPR(10^-5mol/L)for0、1、3、5、7d were detected byRT-PCR in MGC80-3and SH-SY5Y.Results:1.The growth of ECA-109、MGC80-3、PANC-1、HeLa and SH-SY5Y cells wereinhibited after the treatment with ATPR(10^-4、10^-5、10^-6、10^-7、10^-8、10^-9mol/L)in vitro. The inhibitory effect enhanced when the dose increasing. The phanerodepressant effect appeared at48hours, but it is significanter after72hours.Cell morphous was observed to be mature in inverted microscope after thetreatment with ATPR(10^-5、10^-6、10^-7、10^-8、10^-9mol/L) in vitro. SCC of ECA-109decreased（P<0.05）; ALP and LDH activity of MGC80-3suppressant（P<0.05）;PANC-1with the content of CA199decreased; CA125of HeLadecreased（P<0.05）; while the expression of NSE of SH-SY5Yincreased（P<0.05）. G0/G1-phase cells were significantly increased whileS-phase cells were decreased with the elevation of ATPR(10^-5、10^-6、10^-7、10^-8、10^-9mol/L) in vitro. Cell cycle progression was blocked in the G0/G1-phase.2. Compared with the control group, the expression of RARα、RARβ wereincreased in MGC80-3; The expression of RARα and RARβ were increasedwhile the expression of RXRα was decreased in SH-SY5Y cells after treat withATPR(10^-5mol/L) for72h.3. The expression of RARα、RARβ、RXRγ were increased in MGC80-3; Theexpression of RARα and RARβ were increased while the expression of RXRαwas decreased in SH-SY5Y cells after treat with ATPR(10^-5mol/L) for0、1、3、5、7d. Conclusion:1. ATPR(10^-4、10^-5、10^-6、10^-7、10^-8、10^-9mol/L) could inhibit the proliferationand induce differentiation tumor cells in some degree in vitro. But thedifferentiation of PANC-1is still need further study.2. Receptors RARα、RARβ and RXRγ may had closely relation of growthinhibition and differentiation treat with ATPR on MGC80-3gastric cancer cellline. Receptors RARα、 RARβ and RXRα may had closely relation of growthinhibition and differentiation on SH-SY5Y cells. But their exact relationshipsstill need to be further confirmed.