4-氨基-2-三氟甲基苯基维甲酸酯对MCF-7细胞增殖和分化的影响及其机制研究

摘要

目的 研究4-氨基-2-三氟甲基苯基维甲酸酯(4-amino-2-trifluoromethyl-phenyl retinate,ATPR) 对人乳腺癌MCF-7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MCF-7细胞后,绘制细胞生长曲线,分析细胞增殖情况;瑞氏-吉姆萨染色法观察细胞形态学改变;酶联免疫法(ELISA法)检测粘蛋白 (mucin 1,MUC-1) 活性;RT-PCR法检测维甲酸受体(RARα、RARβ、RARγ)、维甲酸受体诱导基因1 (RRIG1)、雌激素受体(ERα、ERβ) mRNA的表达;Western blot法检测RARα、RARβ、RARγ蛋白的表达.结果 ATPR明显抑制MCF-7细胞增殖,且随浓度和时间增加而逐渐增强;镜下观察ATPR作用72 h后MCF-7细胞形态趋向正常细胞分化;ELISA结果显示ATPR明显降低MCF-7细胞培养上清MUC-1浓度(P<0.05);ATPR作用MCF-7细胞72 h后,RARβ、RRIG1、ERβ表达增强(P<0.05),RARγ表达下调(P<0.05),RARα和ERα表达则无明显变化.结论 ATPR可明显抑制MCF-7细胞增殖并诱导其分化程度增高,其机制可能与调节维甲酸受体和雌激素受体平衡,并上调RRIG1表达有关.%Aim To investigate the effects of 4-amino-2-trifluoromethyl-phenyl retinate ( ATPR ) on the proliferation and differentiation of human breast cancer MCF-7 cells and its possible mechanisms. Methods MCF-7 cells were treated with ATPR at different concentrations , cell proliferation was assessed by cell growth curve. Morphologic changes were observed by inverted phase contrast microscope following Wright-Giemsa staining. The differentiation maker mucin 1 ( MUC-1 ) was measured by enzyme linked immunosor-bent assay ( ELISA ). The mRNA expression of retinoic acid receptors ( RARα , RARβ , and RARγ ) , estrogen receptors ( ERα and ERβ ), and retinoid receptor-induced gene-1 ( RRIG1 ) in different groups were detected by Reverse Transcriptase-Polymerase Chain Reaction ( RT-PCR ). The protein expression of retinoic acid receptors ( RARα, RARβ, and RARγ ) were detected by Western blot. Results The proliferation of MCF-7 cells was distinctly inhibited by treatment with ATPR and was in a dose- and time-dependent manner.MCF-7 cells treated with ATPR showed higher degree of mature and differentiation. Compared with that of the control and vehicle ( alcohol ) group, the MUC-1 concentration of cells treated with ATPR was significantly decreased ( P < 0. 05 ), and the expression of RARβ, RRIG1 and ERβ were remarkably increased ( P < 0. 05 ) while the expression of RARγ was notably decreased ( P < 0. 05 ) in cells treated with ATPR. There was no significant difference of RARα and ERα expression between groups. Conclusions ATPR could significantly inhibit the proliferation and induce differentiation of human breast cancer MCF-7 cell, the mechanism of which might be involved with its function in mediating the balance of retinoic acid receptors and estrogen receptors and increasing the expression of RRIG1.