The Effect Of Beclin1 On STX17-Mediated Autophagy In Cervical Cancer SiHa Cells

Objective:To explore the expression level of autophagy-lysosomal key protein STX17 in cervical cancer SiHa cells infected with human papillomavirus 16,and to explore the effect of Beclin1 on autophagy-lysosomal key protein STX17-mediated autophagy in cervical cancer SiHa cells.Methods:The m RNA and protein expression levels of STX17 in SiHa cells and C33 A cells were detected by quantitative real-time polymerase chain reaction and western blotting.Beclin1 was stably overexpressed in SiHa cells by lentiviral transfection technology.SiHa cells in blank control group,SiHa-NC cells in negative control group,and SiHa-Beclin1 cells in overexpression group were set up.Quantitative real-time polymerase chain reaction and western blotting were used to detect the m RNA and protein expression levels of STX17 in SiHa,SiHa-NC and SiHa-Beclin1 cells.The proliferation activity of SiHa,SiHa-NC and SiHa-Beclin1 cells was detected by CCK8 assay;the migration ability of C33 A,SiHa,SiHaNC and SiHa-Beclin1 cells was determined by cell scratch assay.Graphpad Prism 9.0statistical software was used for statistical analysis,t-test was used when two independent samples were homogenous of variance,LSD-t test was used for multiple comparisons between multiple groups,and the test level was α=0.05.Results:(1)The results of quantitative real-time polymerase chain reaction showed that compared with C33A cells,the relative expression level of STX17 m RNA in SiHa cells was lower than that in C33A cells(P<0.0001);compared with SiHa-NC cells,the relative expression levels of STX17m RNA in SiHa-Beclin1 cells was higher than that of SiHa-NC cells(P=0.0005);compared with SiHa cells,the relative expression level of STX17 m RNA in SiHa-Beclin1 cells was higher than that in SiHa cells(P=0.0005).(2)Western blotting results showed that compared with C33 A cells,the relative expression level of STX17 protein in SiHa cells was lower than that in C33 A cells(P=0.0087);compared with SiHa-NC cells,the relative expression level of STX17 protein in SiHa-Beclin1 cells was higher(P=0.0236);compared with SiHa cells,the relative expression level of STX17 protein in SiHa-Beclin1 cells was higher(P=0.0330).(3)CCK8 experiment results showed that compared with SiHa-NC cells,there was no significant difference between SiHa-Beclin1 cells and SiHa-NC cells after 24 hours of stimulation(P=0.5571);after 48 hours of stimulation,the proliferation activity of SiHa-Beclin1 cells reduced(P=0.0160);after 72 hours of stimulation,the proliferation activity of SiHa-Beclin1 cells was significantly weakened(P< 0.0001);after 96 hours of stimulation,the proliferation activity of SiHa-Beclin1 cells was significantly decreased(P< 0.0001).Compared with SiHa cells,there was no significant difference between SiHa-Beclin1 cells and SiHa cells after 24 hours of stimulation(P=0.5341);after 48 hours of stimulation,the proliferation activity of SiHa-Beclin1 cells reduced(P=0.0101);After 72 h,the proliferation activity of SiHa-Beclin1 cells was significantly weakened(P<0.0001);after 96 h of stimulation,the proliferation activity of SiHa-Beclin1 cells was significantly decreased(P<0.0001).(4)Cell scratch experiment results indicated that compared with C33 A cells,the cell migration capacity of SiHa cells was significantly enhanced at 24 h and 48h(P<0.0001);compared with SiHa-NC cells,the cell migration capacity of SiHa-Beclin1 cells was reduced at 24h(P=0.0198),the migration ability of cells at 48 h was significantly weakened(P=0.0017);compared with SiHa cells,the migration ability of SiHa-Beclin1 cells was attenuated at 24h(P=0.0239),and the migration ability of cells at 48 h was significantly decreased(P<0.0001).Conclusion:(1)The expression level of STX17 was lower in cervical cancer SiHa cells,suggesting that autophagy is inhibited.(2)After Beclin1 overexpression,the expression level of STX17 increased in cervical cancer SiHa cells,suggesting that Beclin1 promotes STX17-mediated autophagy in cervical cancer SiHa cells.