摘要:
目的:观察4-氨基-2-三氟甲基苯基维甲酸酯(4-anino-2-trifluoromethyl-phenyl retimate, ATPR)对人乳腺癌细胞株MDA-MB-231抑制增殖诱导分化作用,探讨其可能的作用机制。方法体外培养人乳腺癌细胞株 MDA-MB-231, MTT检测细胞增殖,绘制细胞生长曲线,瑞氏-吉姆萨染色观察细胞形态变化,酶联免疫法检测粘蛋白MUC-1活性,流式细胞术检测细胞周期,实时荧光定量PCR法和Western blot法检测维甲酸受体(retinoic acid receptors,RAR) RARα、RARβ、RARγ和维甲类受体( retinoid X receptors, RXR ) RXRα、RXRβ、RXRγ基因和蛋白的表达。结果 ATPR 能够抑制MDA-MB-231细胞的增殖,具有浓度-时间依赖性,染色后镜下观察 MDA-MB-231细胞生长密度降低,形态趋于正常。ELISA结果显示,ATPR作用后明显降低MDA-MB-231细胞培养上清中MUC-1的浓度;流式细胞术结果显示,MDA-MB-231细胞中G0/G1期表达量增加,S期表达量减少,细胞阻滞在G0/G1期比例增加。 q-RT-PCR和Western blot结果显示, ATPR作用后, RARγ的mRNA和蛋白表达水平降低, RXRs mRNA和蛋白水平无明显变化。结论 ATPR可以抑制人乳腺癌细胞株MDA-MB-231增殖并诱导其分化,其机制可能与RARγ的表达有关。%Aim To investigate the effect of 4-Amino- 2-Trifluoromethyl-Phenyl Retinate on human breast cancer cells MDA-MB-231 and the possible mecha-nisms. Method Human breast cancer MDA-MB-231 cells were incubated with different concentrations of ATPR in vitro. MTT assay was performed to measure the proliferation of MDA-MB-231 . Cell growth curves were made by counting cells and morphologic changes were observed by Wright-Giemsa staining. The differ-entiation marker mucin-1 ( MUC-1 ) was measured by enzyme linked immunosorbent assay ( ELISA ) . Cell cycle was examined by Flow cytometry ( FCM ) . The expression of retinoic acid receptors ( RARs) and reti-noid X receptors ( RXRs ) were detected by Western blot and Quantitative real-time PCR (q-RT-PCR),re-spectively. Results Compared with solvent group, ATPR could inhibit the proliferation of MDA-MB-231 cells in a time-and dose dependent manner and induce the maturing and normality of morphology. The express of MUC-1 was significantly decreased, and the progres of cell cycle was blocked in the G0/G1-phase. The ex-pression of RARγ was decreased. Conclusions AT-PR could inhibit proliferation and induce differention of MDA-MB-231cells, it′s associated with RARγ.