Resveratrol Regulated ARHI Expression In Bladder Cancer And Its Potential Influence In JAK-STAT3 Signaling

Background and Objectives:Bladder cancer,as the most common malignancy of urinary tract,is the fourth most common cancer and is the ninth leading cause of death from cancer among men in the United States.The ratio of men to women that develop bladder cancer is approximately 3:1.It is estimated that bladder cancer will account for 74,960 new cases of cancer and 15,580 cancer-related deaths in the United States during 2014.In China during 2014,the incidence of bladder cancer was 6.61/100,000,the mortality rate was 2.53/100,000,showing an increasing trend year by year.About 60% of patients with bladder cancer are 65 years old.Bladder cancer is a malignant tumor of the urinary tract which develop in the bladder mucosa.Cancer types are mainly divided into urothelial cancer,squamous cell carcinoma and adenocarcinoma.Approximately 90% of bladder cancers were urinary tract epithelial cancer(transitional cell carcinoma,TCC),3%~7% were squamous cell carcinoma,and 2% were adenocarcinoma.Around 70%-85% of TCCs are superficial,also known as non-muscle invasive bladder cancer(NMIBC),including Ta,T1,Tis phase.Transurethral resection of the bladder tumor(TURBT)is the standard treatment for NMIBC in clinical.However,50%~70% of patients will relapse and 10%~30% of the recurrent tumors will develop into muscle invasive bladder cancers.The remaining 15%~30% of transitional cell carcinoma are muscle invasive bladder cancer,including T2-T4 phase.The prognosis of patients with muscle invasive bladder cancer is very poor,nearly 50% of the patients will die within 5 years.In order to prevent and reduce the recurrence of bladder cancer and its further deterioration,chemotherapy and immunotherapy are widely used for treatments of non-muscle invasive bladder cancer.However,such treatments often have limited efficacy and adverse side effects.Therefore,it is significant to seek more effective and non-toxic approach for treatment of transitional cell carcinoma.Resveratrol(Res),a natural polyphenolic compound,has high medicinal value and non-toxic.Which has been found in berries,grapes,peanuts and so on.Resveratrol possesses multiple biological activities,such as anti-aging,anti-inflammatory,antitumor,antioxidant.Present studies have demonstrated that resveratrol has inhibitory effect on ovarian cancer,gastric carcinoma,medullary wilms tumor,breast cancer and other cancer cell growth.So resveratrol is widely concerned all of the world whether it can be used as a new anticancer drug.However,the anti-tumor mechanisms of resveratrol are varied.For instance,regulating Wnt,JAK/STAT3,Notch signaling pathways.Our studies have already confimed that resveratrol can inhibit the growth of bladder cancer cells by regulating the JAK/STAT3 signaling pathway.ARHI(aplasia ras homologue member I)is a kind of maternal imprinted gene,which is located on human chromosome 1p31 and encodes a 26 KDa protein.According to the studies,ARHI can play the role of tumor suppressor in the process of tumor through many ways.including regulating JAK/STAT3 signaling pathway and cell autophagy.However the expression of ARHI and its related tumor suppressor mechanism in bladder cancer has not been reported.Therefore,this study will be based on the study of our lab,to further explore the expression of ARHI and ARHI-related tumor suppressor pathway in bladder cancer.Methods:Using immunohistochemical method for detecting the expression change of ARHI between bladder cancer tissue and adjacent normal tissue.Different sensitivities of human bladder cancer cell line EJ and T24 was cultured and collected after resveratrol treatment for 48 hours.And then RT-PCR,Western-blotting and ICC were used to detect the expression of ARHI and its related protein factors.Results:1)IHC results showed that in the Bladder cancer tissues the positive rate of ARHI is 64%(16/25),and the negative rate is 36%(9/25),and in the adjacent relatively normal tissues of cancer the positive rate of ARHI is 100%(5/5).From the view of expression level,the expression of ARHI in the Bladder cancer are negative(36%)and weakly positive(64%),and in the adjacent tissues of cancer,the expression of ARHI are weakly positive(20%)and intense positive(80%),Meanwhile,there are statistical significance(P<0.01).2)After treated by resveratrol for 48 hours in Bladder cancer,the expression level of ARHI was up-regulated.3)After treated by resveratrol,p-STAT3,VEGF and c-myc were down-regulated at the same time in EJ and T24 cell lines.Besides,nuclear translocation of p-STAT3 was inhibited apparently.Conclusions:1)Compared with the paracancerous tissue,the expression of ARHI declined in the Bladder cancer tissues,which might be associated with the occurrence of tumors.2)Resveratrol can improve ARHI expression in bladder cancer,which showed that resveratrol may inhibit the growth of Bladder cancer by regulating the expression of ARHI.3)After treated by resveratrol in EJ and T24 cell lines,the expression of ARHI was up-regulated,but p-STAT3,VEGF and c-myc were down-regulated.It showed that resveratrol may decrease nuclear translocation of p-STAT3 via up-regulating the expression of ARHI,then influence the expression of downstream proteins like VEGF and c-myc.