RUNT-associated Transcription Factor 2 Interactes With Stearyl CoA Desaturase 1 And Activates Wnt/β-catenin Signaling Pathway To Promote The Progression Of Clear Cell Renal Cell Carcinoma

Introduction:Renal cell carcinoma(RCC)is considered as a kind of common malignant tumor from urinary system,after bladder cancer and prostate cancer on aspect of incidence,with a proportion of 4.18% in whole adult malignancy and of 21.82% in urinary malignancy.RCC accounts for about 300,000 new cancer cases all over the world annually,contributing to almost 100,000 deaths in every year.Clear cell renal cell carcinoma(cc RCC),as the most common histologic subtype,is a tumor regarded as originate from epithelial cells in the proximal convoluted tubule of the nephron.Eventually,around50% of cc RCC patients would develop metastases.Although in last few years the systemic treatment of metastatic cc RCC has come a long way,the patients’ 5-year survival rate with metastatic cc RCC turns out to be lower than 10%.Thus,for improvement of the prognosis of patients with cc RCC,effective biomarkers along with brand-new therapeutic targets are needed to recognize.Previous studies have demonstrated that Runt-associated transcription factor 2(RUNX2)is a main transcription factor in osteoblast and chondrocyte.RUNX2 is related to a variety of tumors,particularly tumor invasion and metastasis,while the expression and molecular mechanisms of RUNX2 in cc RCC keep to be determined.Stearyl Co A desaturase 1(SCD1),an endoplasmic reticulum fatty acid desaturase,transfers saturated fatty acids to monounsaturated fatty acids,being expressed highly in numerous malignancies.Abnormal activation in Wnt/β-catenin signaling pathway is taken as being able to increase the malignancy of human cancers of all kinds.There is evidence that Wnt/β-catenin signaling pathway plays a dominant role in cc RCC growth together with metastasis.Although prior research has indentified a few methods that could be used in the metastatic cc RCC,such as tyrosine kinase inhibitor and m TOR inhibitor,new potential biomarkers and therapeutic method are remained to be explored.In summary,clinical information collection,biological information statistics,and vivo experiments were used to explore the role of RUNX2 and SCD1 in cc RCC and how they promote the occurrence and development of cc RCC through the Wnt/β-catenin signaling pathway.Methods:1.The expression of RUNX2 and SCD1 in 120 pairs of cc RCC tissues was detected After surgical resection,120 pairs of paired cc RCC tissues were collected,and finally histamin was extracted through tissue grinding and cracking.Finally,the expression levels of RUNX2 and SCD1 in 120 pairs of paired cc RCC tissues were detected by Western blot.2.The effect of RUNX2 on the proliferation and invasion of cc RCC cells was tested in vivo.The target gene RUNX2 was knocked out and overexpressed in cancer cells by lentivirus transfection,and the final knockdown efficiency and overexpression were determined.Migration,invasion and proliferation experiments were conducted to verify the effects of RUNX2 on the migration,invasion and proliferation of cc RCC cells.The effects of RUNX2 on the Wnt/β-catenin signaling pathway were detected by Western blot.3.Explore the regulatory mechanism of RUNX2 and SCD1The protein level of SCD1 was detected by Western blot in the stable RUNX2 knockout cells and control cells.IP assay was used to verify the binding of RUNX2 and SCD1 proteins.4.The effect of RUNX2 on the proliferation of cc RCC cells through SCD1 was verified by the response experiment.The recurrent expression of SCD1 was detected by transwell assay,EDU cell proliferation assay and clonogenesis assay.Results:1.As revealed by Western blot,protein level of RUNX2 and SCD1 was prominently higher in tumor tissues than that in the adjacent normal tissues.Next,the interaction between the protein expression of RUNX2 and SCD1 in 120 cases of cc RCC was tested using a linear correlation analysis,and the expression of RUNX2 was correlated positively with protein level of SCD1.2.RUNX2 promotes the development of cc RCC through the Wnt/β-catenin signaling pathwayTranswell,EDU proliferation experiments and clonal formation experiments showed that RUNX2 knockdown reduced the function of migration,invasion and proliferation,and decreased the expression of β-catenin protein.Meanwhile,when RUNX2 was overexpressed,the corresponding migration,invasion and proliferation functions were increased,and the expression level of β-catenin protein was increased.3.RUNX2 binds to SCD1 protein and affects the ubiquitination degradation of SCD1proteinWestern blot results showed that the protein level of SCD1 decreased after RUNX2 knockout.IP assay showed that RUNX2 binds to SCD1 proteins,and the half-life of SCD1 decreases correspondingly after RUNX2 is knocked out.4.SCD1 can partially restore the role of RUNX2 in cc RCC progression After SCD1 was overexpressed in cancer cells,Western blot results showed that the expression of β-catenin protein was increased,and the corresponding function of proliferation,invasion and migration was reversed.Conclusions:RUNX2 and SCD1 play an important role in the development of cc RCC.RUNX2 binds to SCD1 proteins and promotes the progression of cc RCC through the Wnt/β-catenin signaling pathway.Moreover,RUNX2 can maintain the stability of SCD1 protein by affecting the ubiquitination degradation of SCD1.