The Effect Of SOCS1 On The Biological Behavior Of Laryngeal Cancer Cells Through The Activation Of Autophagy By Acetylation Modification Of NAT10

Objective To demonstrate the expression of NAT10,SOCS1 and autophagy and apoptosis-related molecules in laryngeal cancer cells,to elucidate in depth the activation of autophagy after NAT10-mediated SOCS1 acetylation,and to further analyze the correlation between NAT10 and the autophagic pathway.Methods Western Blot was used to detect the background expression of NAT10 in multiple strains of laryngeal cancer cells,and the laryngeal cancer cell line with the highest background expression was selected for subsequent experiments.Lentivirus infection was used to knock down NAT10,and the proliferation,migration and apoptosis levels of laryngeal cancer cells were detected by CCK8,cell scoring and apoptosis assay.The expression levels of autophagy and apoptosis-related molecules(P62,LC3 BII,Bcl-2,BAX,c-Caspase3)and the expression level of suppressor of cytokine signalling(SOCS1)were detected using Western Blot.Pathway activator assay was performed using autophagy signalling pathway activator(MG132),and changes in the expression levels of SOCS1 and autophagy signalling pathway proteins(P62,LC3BII)and apoptosis levels were detected using Western Blot and flow cytometry to confirm whether NAT10 is involved in the autophagic pathway.Results Western Blot results showed that compared to HOK cells,NAT10 expression was decreased in laryngeal cancer cells Hep2,while expression was increased in TU686 and TU212 cells,with more pronounced upregulation in TU686,and TU686 cells were selected for subsequent experiments.CCK8 results showed that compared to the control group(sh Ctrl group),NAT10 knockdown group(sh NAT10 group)TU686 cells had significantly reduced proliferation ability.The results of cell scratch assay showed that the migration ability of sh NAT10 group was significantly reduced compared to sh Ctrl group.The results of apoptosis assay showed that the number of apoptosis was significantly increased in the sh NAT10 group compared to the sh Ctrl group.western Blot results showed that the expression of SOCS1 protein was significantly increased,the expression of autophagy-related protein P62 was significantly increased and the expression of LC3 B II protein was significantly decreased in the sh NAT10 group compared to the sh Ctrl group;the expression of apoptosis proteins Bax and c-C aspase3 expression was significantly increased,while anti-apoptotic protein Bcl-2 protein expression was significantly decreased.After pathway activator experiments using MG132,Western Blot results showed that SOCS1 protein expression was significantly higher in the sh NAT10 group compared to the sh Ctrl group,while autophagy-related protein P62 expression was significantly higher,while LC3 B II protein expression was significantly lower.In contrast,after knockdown of NAT10 with the addition of autophagy activator MG132,SOCS1 and P62 protein expression was reduced and LC3 B II protein expression was significantly increased in the sh NAT10 group + MG132 group compared to the sh NAT10 group.Flow cytometry results showed that the number of apoptotic cells was significantly reduced after the addition of MG132 compared to the sh NAT10 group.The results demonstrated that MG312 could act on the autophagic signalling pathway involving NAT10,which plays a key role in the development of laryngeal carcinoma by mediating the ac4 c acetylation modification of SOCS1 and activating the intracellular autophagic pathway leading to apoptosis.Conclusion In laryngeal cancer cells,the RNA ac4 C acetylation modifying enzyme NAT10 was highly expressed,and NAT10 induced autophagy activation by mediating ac4 c acetylation modification of SOCS1 m RNA and suppressing SOCS1 protein levels.