Wang Hao, Gui Shu-Yu, Chen Fei-Hu, Zhou Qing, Wang Yuan
Department of Respiratory Medicine, the First Affiliated Hospital, Hefei, Anhui, China E-mail :
Asian Pac J Cancer Prev. 2013;14(12):7265-70. doi: 10.7314/apjcp.2013.14.12.7265.
The present study was designed to investigate the probable mechanisms of synthetic retinoid 4-amino-2-tri-fluoromethyl-phenyl ester (ATPR) inhibition of the proliferation and migration of A549 human lung carcinoma cells.
After the A549 cells were treated with different concentrations of ATPR or all-trans retinoic acid (ATRA) for 72 h, scratch-wound assays were performed to assess migration. Immunofluorescence was used to determine the distribution of CAV1 and RXRα, while expression of CAV1, MLCK, MLC, P38, and phosphorylation of MLC and P38 were detected by Western blotting.
ATPR could block the migration of A549 cells. The relative migration rate of ML-7 group had significantly decreased compared with control group. In addition, ATPR decreased the expression of a migration related proteins, MLCK, and phosphorylation of MLC and P38. ATPR could also influence the expression of RARs or RXRs. At the same time, CAV1 accumulated at cell membranes, and RXRα relocated to the nucleus after ATPR treatment.
Caveolae may be implicate in the transport of ATPR to the nucleus. Change in the expression and distribution of RXRα may be implicated in ATPR inhibition of A549 cell proliferation. The mechanisms of ATPR reduction in A549 cell migration may be associated with expression of MLCK and phosphorylation of MLC and P38.
本研究旨在探讨合成类视黄醇4-氨基-2-三氟甲基苯基酯（ATPR）抑制A549人肺癌细胞增殖和迁移的可能机制。
用不同浓度的ATPR或全反式维甲酸（ATRA）处理A549细胞72小时后，进行划痕试验以评估细胞迁移。采用免疫荧光法测定小窝蛋白1（CAV1）和视黄酸X受体α（RXRα）的分布，同时通过蛋白质免疫印迹法检测CAV1、肌球蛋白轻链激酶（MLCK）、肌球蛋白轻链（MLC）、P38的表达以及MLC和P38的磷酸化水平。
ATPR可阻断A549细胞的迁移。与对照组相比，ML-7组的相对迁移率显著降低。此外，ATPR降低了迁移相关蛋白MLCK的表达以及MLC和P38的磷酸化水平。ATPR还可影响视黄酸受体（RARs）或视黄酸X受体（RXRs）的表达。同时发现，经ATPR处理后，CAV1在细胞膜上聚集，RXRα重新定位到细胞核。
小窝可能参与了ATPR向细胞核的转运。RXRα表达和分布的变化可能与ATPR抑制A549细胞增殖有关。ATPR降低A549细胞迁移的机制可能与MLCK的表达以及MLC和P38的磷酸化有关。
