The Effect Of Deacetylase SIRT6 On The Proliferation,Migration Of Renal Cancer Cells

Renal cell carcinoma is one of the most common and fatal tumors of the urinary system.Sirtuins belongs to an evolutionarily highly conserved family of NAD+-dependent deacetylases,which carries a variety of cellular functions and is related to proliferation,DNA repair,mitochondrial energy homeostasis and antioxidant activity.Mammals express seven Sirtuins(SIRT1-7),which have different subcellular localization.Changes in Sirtuins expression are crucial in a variety of diseases,including metabolism-related diseases,diabetes,tumors and aging-related diseases.As a member of the Sirtuins family,there are few studies on SIRT6 in renal cell carcinoma,and the specific mechanism is not clear.Therefore,the purpose of this study is to explore the effect of SIRT6 on the proliferation and migration of renal cell carcinoma cells and whether it can regulate the PI3K/AKT signal pathway in this process.MethodWe embedded sections of renal cell carcinoma and its corresponding paracancerous tissues obtained by surgical resection in patients with renal cell carcinoma.Immunohistochemical staining of SIRT6 was used to determine the expression of SIRT6 in renal cell carcinoma and paracancerous tissues.At the same time,the expression of SIRT6 in human renal cancer cell lines ACHN,786 muro and normal renal tubular epithelial cell HK-2,was detected by Western blotting.In order to determine the effect of interfering SIRT6 on the migration of ACHN cells,we used three kinds of synthetic si RNA fragments to interfere with SIRT6,and selected one of the most efficient si RNA fragments for follow-up experiments.At the same time,WB was used to detect the expression of SIRT6 gene and to determine the effect of SIRT6 on the acetylation of downstream target gene H3K9.The migration ability of ACHN cells was determined by transfection of si RNA,and scratch test and Transwell test were performed on the ACHN 48 hours after transfection of si RNA.At the same time,in order to further explore the effect of SIRT6 on renal cell carcinoma,ACHN cells were treated with OSS＿128167,a specific inhibitor of SIRT6,and the effect of OSS＿128167 on the expression and activity of SIRT6 was detected by WB.At the same time,after ACHN cells were treated with OSS＿128167 for 24 hours,MTT assay was used to detect the effect of OSS＿128167 on the proliferation of ACHN cells.Finally,WB was used to detect the changes of PI3K/AKT signal pathway in ACHN cells treated with OSS＿128167.ResultsThe expression of SIRT6 in renal cell carcinoma was significantly higher than that in paracancerous tissues.At the same time,human renal cancer cell lines ACHN,786 muro and human normal renal tubular epithelial cell HK-2 were selected and the expression of SIRT6 in the cells was detected.The results of Western Blot showed that the expression of SIRT6 in renal cancer cell lines ACHN and 786 Mel O was significantly higher than that in human normal renal tubular epithelial cells HK-2.After transfection of SIRT6 si RNA into ACHN cells,the acetylation level of histone H3K9,the target protein downstream of SIRT6,was significantly increased,while the total protein expression of histone H3 had no significant change.The results of scratch assay showed that the migration ability of ACHN cells was significantly decreased after SIRT6-si RNA transfection,and the migration ability of ACHN cells was significantly decreased after SIRT6 si RNA transfection.Transwell assay showed that the migration ability of ACHN cells decreased significantly after SIRT6 si RNA transfection.The results of Western Blot showed that compared with the control group,the expression of acetylated H3K9 protein increased gradually in ACHN cells with the increase of OSS＿128167 concentration(50 μM,100 μM,200 μM),but had no effect on the expression of SIRT6 protein and H3 total protein.The results of MTT showed that the proliferation activity of renal cancer cell line ACHN decreased significantly with the increase of concentration of OSS＿128167(50 μM,100 μM,200 μM)for 24 hours.After ACHN cells were treated with different concentrations of OSS＿128167(50 μM,100 μM,200 μM),the expression of p-PI3 K and p-AKT protein decreased gradually with the increase of concentration,while the expression of PI3 K and Akt protein did not change significantly.ConclusionSIRT6 may be an unfavorable factor in renal cell carcinoma.The protein can promote the proliferation and migration of tumor cells,and the biological behavior of renal cell carcinoma can be blocked by OSS＿128167,a specific inhibitor of SIRT6.At the same time,we also identified the downstream signal pathway PI3K/AKT which may be regulated by OSS＿128167 in renal cell carcinoma.