Effect Of PRRX1 On Autophagy Of Salivary Adenoidcystic Carcinoma Cells

Objective:To investigate the effect of paired related homeobox1(PRRX1)on autophagy of salivary adenoidcystic carcinoma(SACC)cells and provide a new idea for the study of the mechanism of targeted therapy for salivary adenoidcystic carcinoma.Methods:Salivary adenoidcystic carcinoma cells were transfected with PRRX1 overexpressing lentiviral vector and empty lentiviral vector.Salivary adenoidcystic carcinoma cells were cultured in medium containing purinomycin for 2 weeks.Stable transfected cell lines were screened out.The experimental cells were divided into three groups: PRRX1 overexpressed lentivirus transfection group,empty virus transfection negative control group and untransfected blank control group.Western blot was used to detect the expression level of PRRX1,microtubule-associated protein 2 light chain 3(LC3-Ⅱ)and autophagy associated protein Beclin1 in cells of the three groups.The autophagosome of the three groups of cells was observed by transmission electron microscopy(TEM).The fluorescence intensity of Beclin1 in the three groups of cells was detected by cellular immunofluorescence method.All experiments were repeated for more than 3 times.SPSS17.0 software was used to analyze the data.The results were expressed as mean ± standard deviation.T test was used for comparison between groups.P<0.05 was considered statistically significant.Results:Compared with the blank control group and the negative control group,the expression of PRRX1 protein in the PRRX1 overexpression group group was higher,and the difference was statistically significant(P<0.05).Cells in each group were observed by TEM.It was found that multiple autophagosomes could be observed in the blank control group and the negative control group,while only a few autophagosomes could be observed in the PRRX1 overexpression group.Western blotting results showed that protein expression of autophagy marker(LC3-Ⅱ/Beclin1)was lower in PRRX1 overexpressed group than in blank control group and negative control group(P<0.05).The results of cellular immunofluorescence showed that the fluorescence intensity of Beclin1,the autophagy marker,was lower in the PRRX1 overexpressed group than in the blank control group and negative control group.Conclusion:In this study,we used lentiviral vector transfection technology to construct stable PRRX1 overexpressed salivary adenoidcystic carcinoma cell line,which confirmed that PRRX1 overexpression can inhibit the autophagy of salivary adenoidcystic carcinoma cells.However,the effect on the clinicopathological features and prognosis of salivary adenoidcystic carcinoma cells remains to be further studied.Our study provides a new idea for the study of the mechanism of targeted therapy for salivary adenoidcystic carcinoma from the perspective of autophagy.