CARMA3 Affects Bladder Tumor Growth Via Metabolic Pathway

Introduction: Bladder cancer is the most common malignant tumor of urinary system,and it is the ninth most common malignant tumor in the world.However,due to the recurrence of bladder and the limited treatment,the treatment burden of bladder is higher than that of other cancers.Further understanding of the mechanism of bladder cancer will help to improve the diagnosis and treatment of bladder cancer and benefit more patients.CARMA3 is highly expressed in many types of tumors,and can promote cell proliferation and metastasis.It is generally believed that CARMA3 can activate NF-?B pathway and promote the expression of a series of downstream oncogenes.Therefore,carma3 may promote the growth of bladder cancer and be a potential therapeutic target.Methods: The expression of CARMA3 was detected by PCR in bladder cancer tissues and cells,and the effects of CARMA3 on proliferation,apoptosis,metastasis and stem cell function were verified in bladder cancer cells.RNA sequencing was used to search for potential downstream regulatory molecules of CARMA3,and PCR and immunoblotting were used to verify them.The specific mechanism of action of CARMA3 on CPS1 was confirmed by NF-?B inhibitors and Ch-IP.The expression of CPS1 was detected by PCR in bladder cancer tissues and cells,and the effect of CPS1 on proliferation and apoptosis was verified in bladder cancer cells.The recovery effect of CPS1 on CARMA3 was determined in cell proliferation and apoptosis.Metabonomics of bladder cancer cells showed the effect of CARMA3 on cell metabolism.The effect of CARMA3 on urea cycle pathway through CPS1 was verified by the determination of metabolites in upstream and downstream.The effect of CPS1 and CARMA3 on nucleic acid metabolism was verified by the effect of nucleic acid metabolites on the proliferation and apoptosis of CPS1 and CARMA3 Results: The expression level of CARMA3 was detected by PCR in bladder cancer tissues and cells,and CARMA3 could promote proliferation,inhibit apoptosis,promote migration and invasion,and promote stem cell function in bladder cancer cells.RNA sequencing was used to find the potential downstream regulatory molecule CPS1 of CARMA3,and PCR and Western blot were used to verify the correlation between the expression of CARMA3 and CPS1.NF-?B inhibitor was used to confirm that the change of NF-?B activity can cause the change of CPS1 expression.Ch-IP confirmed that P50 transcription factor can combine with the promoter region of CPS1 to affect the transcription of CPS1.PCR was used to detect the increased expression of CPS1 in bladder cancer tissues and cells,and to verify the function of CPS1 in promoting proliferation and inhibiting apoptosis in bladder cancer cells Overexpression of CPS1 can restore the inhibition of proliferation and promotion of apoptosis induced by knockdown CARMA3 in cell proliferation and apoptosis.Metabonomics of bladder cancer cells shows that CARMA3 mainly affects the urea cycle related pathway in amino acid metabolism.The metabonomics and metabolites of bladder cancer cells confirmed the correlation between the metabolism function of CARMA3 and CPS1.The addition of nucleic acid metabolites can restore the effect of knock down CPS1 and CARMA3 on inhibiting proliferation and promoting apoptosis.Conclusion: CARMA3 is highly expressed in bladder cancer tissues and cells,and can affect bladder cancer cells through cell proliferation and apoptosis,migration and invasion,and stem cell function,and play a role in promoting the growth of bladder cancer cells.CARMA3 can affect MAPK,TGF?,urea cycle and other key pathways,and regulate the expression of CPS1 through NF-?B1,a transcription factor of NF-?B pathway.CPS1 is highly expressed in bladder cancer and can affect the growth of bladder cancer cells through cell proliferation and apoptosis.At the same time,CARMA3 can affect the proliferation and apoptosis of bladder cancer cells through CPS1.CARMA3 can affect the metabolism pathway related to urea cycle,and affect the metabolism pathway of tumor nucleic acid through CPS1.Moreover,CARMA3 and CPS1 can affect the proliferation and apoptosis of bladder cancer cells through nucleic acid metabolism related metabolites.