The Effect Of All Trans Retinoic Acid (ATRA) And Its Derivatives ATPR On Migration Of Gastric Adenocarcinoma Cell Line BGC823and Its Mechanism

Objective To explore the effects of all-trans retinoic acid (ATRA) and its derivatives4-amino-2-trifluoromethyl-phenyl retinate (ATPR) on migration of human gastricadenocarcinoma cell line BGC823and its probable mechanism.Methods After treated with different concentrations of ATRA and ATPR, MTTassay was performed to measure the proliferation of gastric cancer BGC823cells. Lightmicroscope was used to observe morphologic changes. The effects of ATRA and ATPRon the migration of BGC823cells were analyzed by wound healing assay. The effect ofATRA and ATPR on the colony formation rate of BGC823cells was measured by platecolony formation assay. Claudin-18location were detected by immunofluorescence inBGC823cells. The mRNA levels of MLCK were analyzed by Real-time quantitativePCR in BGC823cells. The expression of RARβ, myosin light chain kinase (MLCK)and phosphorylation of myosin light chain (MLC) proteins were detected by westernblot in BGC823cells.Results ATRA can inhibit the proliferation of BGC823cells, but the inhibition rateis low, with the increase of concentration, the inhibitory effect increased gradually. Theproliferation of BGC-823cells was inhibited significantly compared with ATRA group.The relative migration rate of ATPR group had decreased significantly compared withATRA group. Furthermore, Claudin-18located on the cell surface were obviouslyincreased compared with control group, this effect of ATPR were more obvious thanATRA. Real-time qPCR showed that ATRA and ATPR can reduce MLCK mRNA level of BGC823cells, and the effect of ATPR were more obvious than ATRAwhich were statistically significant. In addition, Western blot results showed that ATPRand ATRA could down regulated the expression of MLCK protein, when BGC823cellswere treated by ATPR with48h, the expression of RARβ and phosphorylation of MLCprotein down regulated obviously than treated by ATRA.Conclusions Our data suggest that ATPR had a better inhibition on the proliferationand migration of gastric cancer BGC823cells than ATRA, its probable mechanism wasassociated with the down regulation of expression of MLCK and phosphorylation ofMLC protein involving RARβ/MLCK pathway.