Expression And Function Of CircHIPK3 In Bladder Cancer

Bladder cancer(bladder cancer,BC)is one of the most common malignant tumors in the genitourinary system,which seriously threatens the life and health of the patient.With the improvement of people's living standards,the increasing life expectancy of the population year by year,changes in lifestyle,eating habits and medical standards,coupled with the increasing pollution of the environment,the incidence of bladder cancer in our country is increasing year by year and younger trend.Early diagnosis and treatment of bladder cancer,will help to improve patient survival.At present,most studies focus on the regulation of protein factors or signaling pathways,while less on the post-transcriptional translation and epigenetic studies of genes.There are few reports on the role of non-coding RNA,especially circRNA,in bladder cancer.Circular RNAs(circRNAs)are a group of circular RNAs formed by covalently closing,usually at the 3 'and 5' ends of an RNA molecule.Because circRNAs do not have 5 'or 3' ends,they are resistant to exonuclease-mediated degradation and are presumed to be more stable than most linear RNAs in cells.CircRNA is abundant in eukaryotic cells at a high degree conservative,structurally stable,with a certain degree of organization,timing and disease-specific.Numerous studies show that circular RNA plays an important regulatory role in the development of cancer.It has been found that many circular RNAs have a positive or negative regulatory relationship with their parental genes.CircHIPK3 is abnormally expressed in many kinds of cancerous tissues,but its expression in bladder cancer and its relationship with its parental HIPK3 gene are largely unclear.HIPK3 is a serine / threonine protein kinase that plays a negative regulatory role in apoptosis by phosphorylation of JUN and RUNX2,which may also act by enhancing androgen receptor(AR)-mediated transcriptional activation.JUN,also known as transcription factor AP-1,which mediates XIAP upregulation of Cyclin D1 expression.In this study,by detecting the RNA expression of circHIPK3 and HIPK3 in bladder cancer and adjacent tissues,we determined the expression of circHIPK3 in interstitial tissue and its relationship with the clinicopathological features of bladder cancer.And examined the relationship between HIPK3 and Cyclin D1.Provide a theoretical basis for the role of circHIPK3 in bladder cancer.Objective: To investigate the expression of circular RNA circHIPK3 and its parental gene HIPK3 and its role in bladder cancer.Method:1 the total RNA and protein were extracted from the tissues and the adjacent of the patients with bladder cancer radical surgery.2 Using qRT-PCR to detect the expression of circHIPK3 and HIPK3 in bladder cancer and adjacent tissues,and the results were statistically analyzed.3 Statistical analysis of circHIPK3 and HIPK3 expression and bladder cancer clinicopathological features of the relationship..4 The expression of HIPK3 and Cyclin D1 protein was detected by Western blot.5 Pearson's correlation analysis was performed using Graphpad prism5 to analyze the linear correlation between circHIPK3 and its parental genes HIPK3 and HIPK3 and Cyclin D1,respectively.Results:1 circHIPK3 and HIPK3 mRNA in bladder cancer and adjacent tissuesCompared with the control group,the expression of circHIPK3 in bladder cancer tissues was significantly decreased(P<0.01),while the expression of HIPK3 mRNA in bladder cancer tissues was significantly increased(P<0.05).The differences in the expression of circHIPK3 and HIPK3 in bladder cancer tissues(25.53% and 76.60%,respectively)were statistically significant(P<0.05).2 circHIPK3 and its parental gene HIPK3 mRNA expression in a linear correlation.There was a negative correlation between the expression of circHIPK3 and its parental HIPK3 mRNA in bladder tissues(R=-0.6984,P=0.0004).3 circHIPK3 and HIPK3 expression in clinicopathological features of bladder cancerIt was found that the expression of circHIPK3 and HIPK3 was not significantly different between individuals of different ages and sex(P>0.05),but negatively correlated with the morphological grade of cancer(P<0.05).In TNM clinical stage,the expression of circHIPK3 in T2 and above patients Significant downregulation,HIPK3 expression was higher than the T2 level(P<0.05).The expression of circHIPK3 was significantly down-regulated in the samples of lymph node metastasis,and the expression of HIPK3 was higher than that in non-metastatic individuals(P<0.05).4 HIPK3 and Cyclin D1 were significantly upregulated in bladder cancer tissues.The expression of HIPK3 and Cyclin D1 protein in bladder cancer tissues was significantly up-regulated compared with that in paracancerous tissues,while the expression of ?-actin was not significantly different between the cancer tissues and paracancerous tissues.Western blot results were gray-scale scans and statistical analysis,as shown in Figure 3 B and C,HIPK3 and Cyclin D1 protein levels in bladder cancer increased significantly(P<0.05).5 There was a linear correlation between the expression of HIPK3 and Cyclin D1 in bladder cancer tissues.There was a linear correlation between the expression of HIPK3 and Cyclin D1 protein in bladder tissue(R = 0.4377,P = 0.0472).Conclusion: The expression of circHIPK3 was significantly decreased in bladder cancer(P<0.01),while the expression of HIPK3 mRNA was significantly increased in bladder cancer(P<0.05).Their expression rates were25.53% and 76.60%,respectively.There was a linear negative correlation between the expression of circHIPK3 and HIPK3 in bladder cancer.There was no significant difference in the expression of circHIPK3 and HIPK3 between individuals of different ages and genders,but negatively correlated with themorphological grade of cancer,TNM clinical stage and lymph node metastasis.The protein levels of HIPK3 and Cyclin D1 were up-regulated in cancerous tissues,and there was a linear positive correlation between them in bladder tissues.