The Effect Of HMGB1Inhibitor On The Invision And Migration In Human Salivary Gland Adenoid Cystic Carcinoma Cell Line And Study Of Their Mechanisms

Objective: To observe the effect of HMGB1inhibitor-Sodium Butyrate on the invisionand migration of human salivary gland adenoid cystic carcinoma cell line in vitro and toinvestigate the underlying mechanisms.Methods:1.The cultured ACC-2and ACC-M cells were treated with differentconcentrations of Sodium Butyrate for24h, detected the viability rate of the cells bymethyl thiazolyl tetrazolium(MTT) assay, and observed the morphological changes byordinary optical inverted microscope.2.The drug’s influence on invision and migrationon ACC-2and ACC-M by Transwell experiment.3.Detected the protein and mRNAexpression of HMGB1and TLR4by Western-blot and RT-PCR assay, and the relationshipbetween TLR4expression and HMGB1was investigated.Results:1.Compared with control group, Sodium Butyrate could inhibit the proliferationof ACC-2and ACC-M cells significantly at the concentrations of0.625mM、1.25mM、2.5mM、5mM、10mM, all the effect were in a dose-dependent manner(P<0.05).2.Thefive group Sodium Butyrate could inhibit the invision and migration of ACC-M cells byTranswell experiment(P<0.05), but only2.5mM、5mM and10mM concentrations groupdrugs could inhibit the migration of ACC-2cells. The influence of Sodium Butyrate on theinvision of ACC-2cells have no significant difference(P>0.05).3.The results ofWestern-blot and RT-PCR showed the five group Sodium Butyrate could dose-dependentlydown-regulate the protein and mRNA of HMGB1and TLR4in ACC-M cells(P<0.05),while2.5mM、5mM and10mM concentrations group could down-regulate the protein andmRNA of HMGB1and TLR4(P<0.05). Correlation analysis displayed: reduced expressionof TLR4protein is positively correlated with that HMGB1protein was inhibited, the resultis statistically significant(r=0.810，P<0.05).Conclusion: The expression of HMGB1plays an important role in the biologicalbehavior of SACC in vitro. Inhibiting the expression of HMGB1could reduce the invision and migration of ACC-M, also the invision of ACC-2cells at a high concentration ofSodium Butyrate. TLR4maybe involved in the regulation of HMGB1for SACC cellsbiological behavior in vitro. This study provided a preliminary experimental evidence forthe screening of HMGB1inhibitor analogues in clinic therapy of SACC.