| Background Renal cell carcinoma,accounting for approximately 90% of adult kidney tumors,is one of the most common malignancies in the urinary system.As the most common pathological type of renal cell carcinoma,renal clear cell carcinoma accounts for about 80% of renal cell carcinoma.In terms of the current technical means,the early diagnosis of renal clear cell carcinoma is still difficult for the cells of 30%-50% of patients have already metastasized at the time of diagnosis.Due to the complex biological characteristics of renal clear cell carcinoma,and its insensibility to radiotherapy and chemotherapy,radical or partial nephrectomy now is the main means for the treatment of renal cell carcinoma.Although the surgical result is effective,20%-40% of patients still have local recurrence or distant metastasis after the surgery.Patients both in stage III and IV renal clear cell carcinoma have poor prognoses for the 5-year survival rate after treatment is only 50% and 10%,respectively.Therefore,the study on the development and metastasis mechanism of renal clear cell carcinoma will be beneficial to the clinical diagnosis and treatment of renal clear cell carcinoma.Ketone bodies are the intermediates produced by fatty acids in the decomposition and oxidation of hepatocytes,including acetoacetate,beta-hydroxybutyrate and acetone.The ketone body is produced by the liver and then transported to and utilized in the heart,kidney,brain and skeletal muscles.As an important organ for the utilization of ketone bodies,the kidney has a strong ability of ketone metabolism.And ketone bodies,the important product of metabolism in the body,are closely related to the occurrence and development of tumors.ACAT1,BDH2 and HMGCL have been reported as key genes in ketone metabolism in various tumor studies,but are seldom mentioned in renal clear cell carcinoma.Therefore,this article will explore the expression of key ketone metabolism genes such as ACAT1,BDH2 and HMGCL as well as their biological functions in renal clear cell carcinoma.Objective:To explore the transcriptional expression levels of ACAT1,BDH2 and HMGCL genes in renal clear cell carcinoma,and their regulation on ketone metabolism as well as the effects of abnormal expression of ACAT1,BDH2 and HMGCL on the malignant biological behavior of renal clear cell carcinoma.Methods:1.The relationship between the m RNA transcription levels and clinical parameters of ACAT1,BDH2 and HMGCL in renal clear cell carcinoma is analyzed by bioinformatics method using GCBI platform,TCGA database and GEPIA website.2.The transcription and expression of ACAT1,BDH2 and HMGCL in renal clear cell carcinoma c DNA microarray and renal clear cell carcinoma tissue microarray are verified by real-time fluorescent quantitative PCR and immunohistochemical staining,respectively. 3.The overexpressing ACAT1,BDH2 and HMGCL of exogenous renal clear cell carcinoma cell lines is constructed.What’s more,by conducting CCK-8experiment to draw growth curve as well as doing,2D cell migration assay and3 D cell invasion assay,the biological effects of ACAT1,BDH2 and HMGCL genes on the malignant tumors in the renal clear cell carcinoma are detected.4.The ketone body content in the exogenously overexpressing ACAT1,BDH2 and HMGCL cells of renal clear cell carcinoma cell lines is detected.5.By treating the renal clear cell carcinoma cell line with exogenous ketone body,the effect of exogenous ketone body on the proliferation ability of renal clear cell carcinoma cell line is verified in CCK-8 assay.Results:1.The expression levels of ACAT1,BDH2 and HMGCL genes are down-regulated in renal clear cell carcinoma.By analyzing bioinformatics data,it is discovered that ACAT1,BDH2,and HMGCL are down-regulated in renal clear cell carcinoma tissues compared to that of in the normal kidney tissues.Moreover,the expression levels of ACAT1,BDH2 and HMGCL are related to the survival and prognosis of renal clear cell carcinoma.Furthermore,by verifying the transcription levels of ACAT1,BDH2 and HMGCL genes in renal clear cell carcinoma c DNA microarray with RT-q PCR,it is found that the transcriptional expression levels of ACAT1,BDH2 and HMGCL genes in renal clear cell carcinoma are lower than that of in adjacent tissues,and the difference is of statistical significane.Furthermore,immunohistochemical staining of tissue microarray is used to compare 90 cases of renal clear cell carcinoma and 90 cases of adjacent tissues.And it is found that the expression levels of ACAT1,BDH2 and HMGCL in cancer tissues are significantly lower than that of in adjacent tissues,and the difference is of statistical significance.The results show that the expression of ACAT1,BDH2 and HMGCL genes are down-regulated in renal clear cell carcinoma.Moreover,their expression levels are related to some clinical parameters.2.Exogenous overexpression of ACAT1,BDH2 and HMGCL genes inhibit the malignant biological ability of renal clear cell carcinoma cells.Through the experiment of CCK-8 cell proliferation assay,cell scratch healing assay and Transwell invasion assay,it is found that exogenous overexpression of ACAT1,BDH2 and HMGCL can inhibit the proliferation and invasion of renal clear cell carcinoma as well as the migration ability of exogenous overexpression of ACAT1 and BDH2.It indicates that ACAT1,BDH2 and HMGCL genes can inhibit the malignant biological behavior of renal clear cell carcinoma cells.3.Exogenous ketone body β-hydroxybutyric acid inhibits the proliferation of renal clear cancer cells.Exogenous overexpression of ACAT1 and HMGCL genes can increase the content of ketone body in renal clear cell carcinoma cells by detection of theβ-hydroxybutyric acid detection kit.It is found by CCK-8 detection that the adding of exogenous ketone β-hydroxybutyric acid can inhibit the proliferation of renal clear cell carcinoma cells,and its inhibitory intensity gradually increases with the increase of β-hydroxybutyric acid concentration.This dependence shows that ACAT1,BDH2 and HMGCL genes can alter the production of β-hydroxybutyrate by jointly regulating ketone metabolism,thereby affecting the proliferation of renal clear cell carcinoma.Conclusion:1.The transcription and expression of ACAT1,BDH2 and HMGCL genes are down-regulated in renal clear cell carcinoma,and their expression levels are related to the prognosis of the patients;2.Overexpression of ACAT1,BDH2 and HMGCL can inhibit the proliferation ability and invasion of renal clear cell carcinoma.Overexpression of ACAT1 and BDH2 can inhibit the migration of renal clear cell carcinoma.3.Overexpression of ACAT1 and HMGCL can increase the content ofβ-hydroxybutyrate in renal clear cell carcinoma cells.4.Exogenous ketone body β-hydroxybutyric acid inhibits the proliferation ability of renal clear cancer cells. |
PDF Full Text Request