| Objective:To investigate antitumor effects of salidroside on salivary adenoid cystic carcinoma.Methods:Human salivary adenoid cystic carcinoma cells SACC-2 were selected as objects for in vitro experiment and cultured in vitro for 24 hours,and the CCK-8 method was used to observe the inhibition of salidroside(0.125 ? g / ml,0.25 ? g / ml,0.5 ? g / ml,1 ? g / ml,2 ? g / ml,4 ? g / ml,8 ? g / ml)on SACC-2 cell proliferation and calculate the IC50 value of salidroside;morever,the effects of salidroside on the invasion and migration of SACC-2 by Transwell test and scratch experiments were observed;TUNEL detection was used to reflect the apoptosis of SACC-2 cells;and Western Blot experiment was used to observe changes in the expression levels of autophagy-related protein LC3 and apoptosis-related proteins Bcl-2 and Bax in SACC-2 to further investigate the effects of salidroside at different concentrations on the apoptosis of SACC-2 cells in vitro.In vivo experiments,the subcutaneous tumor model of salivary adenoid cystic carcinoma cells was established in nude mice.The groups were set to saline group,5-Fu group,salidroside group and combination drug group(5-Fu+salidroside)four groups,HE staining,immunohistochemical staining and TUNEL detection were used to reflect the changes of tumor cell morphology,proliferation and apoptosis after salidroside treatment,and to further explore the mechanism of salidroside regulating autophagy-related protein LC3 and apoptosisrelated protein Bcl-2,Bax expression in vivo,and then affecting SACC-2 cell apoptosis.Results:1.CCK-8 experiment showed that gradient concentration of salidroside could inhibit the proliferation and growth of SACC-2 cells to different degrees,and the inhibitory effect gradually increased with the increase of salidroside concentration,and its IC50 value was 5.07 ± 0.91?g / ml.2.Transwell test and scratch test showed that,compared with the control group,salidroside at 5?g / ml could significantly reduce the number of perforated cells,prolong the healing time of the scratch,and reduce the migration and invasion ability of SACC-2 cells.3.TUNEL detection could observe that,with the increase of salidroside concentration,the blue-green fluorescence intensity increased gradually,as well as the apoptosis level of SACC-2 cells.Western Blot experiment showed that compared with the control group and the 2.5ug / ml salidroside group,the expression levels of Bcl-2 and Bax in the 5ug / ml salidroside group were reduced significantly.The ratio of Bcl-2 / Bax and the LC3-II / I increased significantly.4.In vivo studies of HE staining showed that the tumor cells in 5-Fu treated group,salidroside treated group and combined drug group had morphological changes,decreased nuclear heteromorphism,and lighted nuclear staining,in which the combination treatment group had the shallowest nuclear staining and the smallest nuclear atypia;immunohistochemical staining showed that after salidroside or / and 5-Fu treatment,the expression level of Ki-67 in tumor cells decreased,and the cell proliferation ability was weakened.The Ki-67 of the cells in the combination group was the least,with the lowest positive expression level.5.The results of immunohistochemical staining in vivo showed that compared with the saline group,the positive expression level of anti-apoptotic protein Bcl-2 in the combined drug group was significantly reduced(P <0.01),while the positive expression level of pro-apoptotic protein Bax was significantly increased(P <0.05);In addition,compared with the saline group,the expression level of autophagy-related protein LC3 in the combined drug group was also significantly increased(P <0.05).Conclusion:1.Salidroside can inhibit the proliferation of SACC-2 cells.2.Salidroside can inhibit the migration and invasion of SACC-2 cells.3.The inhibitory effect of salidroside on SACC-2 cell proliferation is achieved by promoting apoptosis,and its mechanism of action is related to the mitochondrial pathway of apoptosis and cell autophagy. |
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