Expression And Significance Of Autophagy-associated Protein STX17 In Cervical Cancer SiHa Cells

Objective:(1)To explore the correlation between HPV and autophagy-lysosomal key protein STX17 in cervical cancer SiHa cells.(2)To explore the effect of STX17 on autophagy,proliferation and invasion of cervical cancer SiHa cells.Methods:The mRNA and protein expression levels of STX17 in cervical cancer C33 A cells and SiHa cells were detected by qRT-PCR and Western blot respectively,and their differences in expression were analyzed;After cervical cancer SiHa cells were transfected by lentivirus,the control group was set as SiHa cells without any treatment(Control group),the experimental group consisted of SiHa cells infected with STX17 overexpressing empty vector lentivirus(SiHa-NC group)and SiHa cells infected with STX17 lentivirus over-expressed(SiHa-OE group).The mRNA expression levels of autophagy-lysosome-related factors LAMP-1 and LAMP-2 were detected by qRT-PCR;the protein expression levels of autophagy-lysosome-related factors LAMP-1and LAMP-2 were detected by Western blot;The proliferation activity of SiHa cells was detected by CCK-8 method;Scratch test was used to analyze the migration ability of Siha cells at 24 h and 48h;Graph Pad Prism9 software was used for statistical analysis,and t test was used between two independent samples with normal distribution and homogeneity of variance;one-way ANOVA was used for comparison between multiple groups,and LSD-t test was used for multiple comparison between groups,with α=0.05.Results:(1)qRT-PCR test: Compared with C33 A cell line,the expression level of STX17 mRNA in SiHa cell line was lower(P<0.0001);Compared with the Control group,there was no significant difference in LAMP-2 mRNA expression level in the SiHa-NC group(P=0.1114);Compared with the Control group and the SiHa-NC group,the expression level of LAMP-2 mRNA in the SiHa-OE group was increased(P=0.0044,P<0.0001);(2)Western Blot test: Compared with C33 A cell line,the expression level of STX17 protein in SiHa cell line was decreased(P=0.037);Compared with the Control group,there was no significant difference in the expression levels of LAMP-1 and LAMP-2 in the SiHa-NC group(P=0.996,P=0.5155);Compared with the Control group and the SiHa-NC group,the LAMP-1 protein expression levels in the SiHa-OE group were increased(P=0.0274,P=0.0303);The expression level of LAMP-2 protein was increased(P=0.0109,P=0.0408);(3)CCK-8 test: Compared with the Control group,the proliferation ability of the SiHa-NC group at 24 h,48h,72 h,and 96 h had no significant change(P=0.9984,P=0.9431,P=0.4779,P=0.0872);Compared with the Control group and the SiHa-NC group,the proliferation ability of the SiHa-OE group at 24 h was lower(P=0.6414,P=0.6748),the proliferation ability at 48 h was lower(P=0.0303,P=0.0133),the proliferative ability at 72 h was lower,with statistical difference(P<0.0001,P<0.0001),the proliferation ability at 96 h was lower(P<0.0001,P<0.0001);(4)Scratch test: Compared with the control group,the migration ability of the SiHa-NC group had no significant change at 24 h and 48h(P=0.9998,P=0.7883);Compared with the Control group and the SiHa-NC group,the 24 h migration ability of the SiHa-OE group was lower(P=0.0002,P=0.0002),the migration ability at48 h was lower(P=0.0001,P=0.0004).Conclusion:(1)Compared with C33 A cells,the expression level of STX17 in SiHa cells was lower,suggesting that HPV infection inhibited the fusion of autophagy lysosomes in cervical cancer cells;(2)In SiHa cells overexpressing STX17,the expression of LAMP-1/2 was higher and the cell proliferation and migration were decreased,suggesting that STX17 may down-regulate the expression of LAMP1/2,inhibit the fusion of autophagosome and lysosome,and finally promote the occurrence and development of cervical cancer.