Expression And Significance Of Acetyl Coenzyme A Synthetase 2 In Renal Carcinoma

Renal cell carcinoma(RCC)is one of the most common malignancies of the adult kidney.Among the RCC histological subtypes,clear cell RCC(ccRCC)is the commonest,accounting for more than 80%of cases.Despite recent efforts in multimodal treatment approaches,the prognosis of patients with RCC remains poor,mainly due to delayed diagnosis and a relatively high incidence of metastasis.It is difficult to predict the clinical outcome for RCC because of heterogeneity of the molecular phenotype.Therefore,there is an urgent need to identify novel diagnostic and therapeutic targets in RCC.Acetyl coenzyme A synthetase 2(ACSS2)is widely expressed in many malignant tumors,but the expression and mechanism of acetyl coenzyme A synthase 2 in renal cell carcinoma is still unknown.Renal metastasis accounted for 5%of the total metastatic cancer in the male,accounting for 4%of the female,17%of the patients were diagnosed as locally advanced stage,16%had distant metastasis when diagnosed,and the 5 year survival rate of distant metastasis patients was only 12%.The recurrence rate of the patients received surgical treatment was up to 40%.In addition,renal cancer is not sensitive to chemotherapy and radiotherapy,and is not sensitive to some immune agents,such as INF-alpha and IL-12.However,some molecular or cellular markers have been found in recent years,leading to targeted therapy for renal metastasis,such as RTK and mTOR enzyme inhibitors.The progression of renal cancer has risen from 5 months to 15 months due to the emergence of targeted therapy.However,there are few completely effective treatments,so RCC is still unpredictable and the prognosis is poor.In cancer,the phosphatidylinositol 3 kinase/AKT(PI3K/AKT)pathway is activated by mutation of gene and the number of replicas(CNAs).The catalytic subunit P110 alpha of PI3K is the most common subunit of catalytic subunit.Gene mutation can lead to the activation of PI3K/AKT pathway,and then lead to a series of multiple kinases and multiple branches involved in the multiple reactions,which provide multiple targets in the treatment of tumors.Therefore,inhibition of PI3K,AKT,mammalian rapamycin targeting gene(mTOR)and the activation of other components have became the direction of cancer treatment.Part ? The relationship between expression of ACSS2 and progression,pathological features and prognosis of renal cell carcinomaObjectiveTo study the relationship between expression of ACSS2 and progression, pathological features and prognosis of renal carcinoma.MethodsUsing immunohistochemical method to perform the expression of ACSS2 in 198 human renal cell carcinoma samples;using qRT-PCR,western blotting and other methods to test the expression of ACSS2 in cell lines(HK-2,ACHN,Caki-1,786-0,and 769-P)and tissue samples of 21 cases from patients with renal cell carcinoma at the transcriptional level;using a variety of statistical methods to study the correlation between the expression and prognosis of ACSS2 with renal cell carcinoma.ResultsThe results of immunohistochemistry showed that the expression of ACSS2 in the RCC tissues was higher than para-cancer tissues;qRT-PCR results showed that ACSS2 mRNA was high expression in Caki-1,786-0,769-P cells,low expression was observed in HK-2 and ACHN cells,the expression of ACSS2 mRNA in renal cell carcinoma tissues was higher than that in the adjacent tissues.The results of western blotting showed that the protein expression of ACSS2 in the cancer tissues is higher than that in the para-cancer tissues.Statistical analysis showed that tumors with high ACSS2 expression were more likely to have advanced T stage,advanced AJCC stage,and high UISS risk category shorter RFS and OS.ConclusionThe expression level of ACSS2 in cancer tissues is higher than that in adjacent tissues.The high expression of ACSS2 indicates that tumors with high ACSS2 expression were more likely to have advanced T stage,advanced AJCC stage,and high UISS risk category shorter RFS and OS.Part ? The effect of the expression of ACSS2 on cell proliferation,migration and invasionObjectiveTo determine the effects of ACSS2 on biological behavior such as proliferation,migration and invasion of renal cell carcinoma cells.MethodsLv-siACSS2 and Lv-siNC were transfected into cell lines to study the expression of ACSS2.The MTT,wound healing experiment,and the Transwell test were used to verify the proliferation,migration and invasion ability of the cells after transfection.ResultsLv-siACSS2 and Lv-siNC were transfected into RCC 786-0 and 769-P cell lines,and the expression of ACSS2 at mRNA and protein levels was reduced.MTT results showed that cell proliferation in group Lv-siACSS2 was significantly inhibited compared with group Lv-siNC.The wound healing results showed that the scratch closure of ACSS2 silencing cells was affected compared with the control group.Compared with the Lv-siNC group,the invasion and metastasis of 786-0 and 769-P cells in group Lv-siACSS2 were also inhibited.RT-qPCR and western blotting experiments showed that the expression level of ACSS2 in native ACHN cells was low,and it was not affected by Ad-NC carrier infection.ACHN expression in Ad-ACSS2 infected cells was 3 times higher than that in ACHN cells.MTT experiments showed that the proliferation of ACHN cells in group Ad-ACSS2 was higher than that in group Ad-NC.The wound healing results showed that the migration ability of Ad-ACSS2 cells was increased compared with the control group(Ad-NC).Transwell results showed that the migration and invasion of Ad-ACSS2 cells increased.ConclusionACSS2 promotes the proliferation,invasion and migration of renal cell carcinoma cells.Part ? The effect of ACSS2 on the PI3K/AKT pathway of renal carcinomaObjectiveTo detect the inhibition of PI3K/AKT after ACSS2 silencing.MethodsThe expression of PI3K protein and p-AKT protein was detected by western blotting.ResultsWestern blotting analysis showed that silencing of ACSS2 could significantly inhibit the expression of PI3K and p-AKT proteins in 786-0 and 769-P cells.ConclusionACSS2 may play the role of oncogene in renal cell carcinoma through PI3K/AKT signaling pathway.