The Expression Of β-Adrenergic Receptors And The Effect Of β-Blockers On The Proliferation,Migration And Invasiveness Of CAL-27 Cell Line

Objective: To investigate the expression of β adrenergic receptors in CAL-27 cell line,and explore the effect of β-blockers on the proliferation,migration and invasion of CAL-27 cell line.Methods The primary culture oral epithelial cells were obtained by isolation from oral mucosa and identified by Immunofluorescent assay.Immunohistochemistry and Realtime quantitative PCR(qRT-PCR)were used to detect the expression of β-adrenergic receptor in CAL-27 and oral mucosal epithelial cells(labial mucosa,palatal mucosa,gingival mucosa).The CAL-27 cell viability was detected by CCK-8 assay with different concentrations of propranolol and metoprolol.Transwell and cell migrationwas performed to assess invasion and migration ability affected by different concentrations of propranolol and metoprolol.Results Immunohistochemistry showed that the expression of β AR in CAL-27 cells were higher than that in normal oral epithelial cells.qRT-PCR showed that the expression of β2-AR was lower than that in oral epithelial cells,whereas the expression of β1-AR was higher than that of normal epithelial cells.The results of CCK-8 assay showed that when the drug concentration>50μM,the inhibiting effect of propranolol on cell viability is significant(F=19.550,P<0.05).With the rise in the concentration of two drugs,the inhibition effect of CAL-27 cells activity is increasing(P<0.05).The result of scratch-wound assay showed that the migration rate of CAL-27 cells was statistically significantly decreased when the drug concentration was not lower than 10μM(F=7.83,P<0.05).Propranolol was more resistant to migration of CAL-27 cells.With the rise in the concentration of both propranolol and metoprolol,the inhibition effect of migration is increasing(P<0.05).The result of transwell assay showed that the invasion rate of CAL-27 cells was significant decreased when the drug concentration was not lower than 10μM(F=16.11,P<0.05).Propranolol was more resistant to invasion of CAL-27 cells.With the rise in the concentration of two drugs,the inhibition effect of invasion is increasing(P<0.05).Conclusions The expression of mRNA of β1-AR in CAL-27 cells is higher than that of oral epithelial cells,and the expression of mRNA of β2-AR is lower than that of oral epithelial cells.Low concentration of propranolol and metoprolol have no significant effect on the proliferation of CAL-27 cells,but decrease the migration and invasion ability of CAL-27 cells.