| Objective:With the development of social and economic development,the incidence of type 2diabetes is increasing year by year,and the incidence and mortality of bladder cancer in the world is also increasing.Type 2 diabetes will increase the incidence and mortality of bladder cancer.Type 2 diabetes is an independent risk factor for bladder cancer.Although there are many common risk factors between type 2 diabetes and bladder cancer,its internal mechanism is still unclear.Therefore,it is urgent to find out the reason behind the relationship between diabetes and bladder cancer.Gene chip is a common technique which can be used to detect differential gene expression in different states.The purpose of this study was to detect bladder cancer cells under high and low glucose conditions.Differential gene expression,screening target genes for verification,and through further cytological experiments to understand its possible mechanism,understand the molecular mechanism of type 2 diabetes prone to cancer,help to achieve early diagnosis,provide experimental basis to prevention of bladder cancer and targeted treatment.Methods:1.Gene chip study: bladder cancer T24 cells were cultured in 5.5mmol/L(low glucose)and 25mmol/L(high glucose)glucose concentration,respectively.Affymetrix human Clariom-D Hu was used.Microarray was used to detect differential expression of Circ RNAs,Lnc RNA,mi RNA and m RNA systematically.with multiple of difference ≥ 2 Target genes were screened by bioinformatics targeting and gene changes associated with high glucose.2.Population validation: we reviewed and collected the medical data of all newly diagnosed patients with bladder cancer in the urology department of the second Hospital of Tianjin Medical University.46 patients with bladder cancer were divided into two groups according to whether or not they were complicated with diabetes mellitus: bladder cancer alone group and bladder cancer complicated with diabetes group;At the same time,we collected the clinical data of 23 bladder cance patients before and after with diabetes mellitus,and all the patients were examined by TRIB3 immunohistochemical staining.Correlation analysis of clinicopathological features.3.cell experiment: T24,EJ cells with high glucose and low glucose were tested clone formation test,We used MTT,transwell to detect proliferation,migration level,used RT-PCR,WB to detect the high glucose and low glucose state of T24,EJ cells TRIB3 m RNA and protein level,MTT,Transwell was used to detect the proliferation and migration level of T24 and EJ cells before and after transfection with TRIB3 si RNA.Further study was made on the role of TRIB3 in promoting the proliferation and migration of T24 cells through MAPK/ERK1/2 pathway.Results:1.Among the above genes,18922 genes were up-regulated and 16776 genes were down-regulated.The difference of Circ RNAs,Lnc RNA,mi RNA,m RNA was3203,9355,1571 and 4792 respectively.The biological processes of differential genes mainly focus on endoplasmic reticulum stress,protein transcription,DNA replication and so on.Differential gene cell composition mainly concentrated on the cell surface,cytoplasm,mitochondria and so on.Differential gene molecular function is mainly in fibroblast growth factor receptor binding,white activity,protein binding and other aspects.The differential gene signaling pathway is enriched in the NF-KB,TNF,nod receptor signaling pathway and so on.The signal pathway changes caused by differential gene changes are mainly concentrated in infectious diseases,immune diseases,and neoplastic diseases.TRIB3 was selected as the research object by screening.2.The bladder cancer complicated with diabetes mellitus group was higher than BMI(p=0.024),the number of patients with hypertension was more(p=0.036),the hemoglobin was lower,the fasting blood glucose was higher(P < 0.01),the tumor stage was higher(P < 0.006),the tumor grade was higher(p=0.040),and the TRIB3 was higher(P < 0.001),and there was a positive correlation between TRIB3 and tumor grade and stage(P < 0.016).The higher expression of TRIB3 in bladder cancer patients after than before diabetes mellitus(p=0.032),the tumor stage and grade were higher(P < 0.01),and the fasting blood glucose was higher(P < 0.01).3.The clone formation,proliferation and migration of EJ cells in T24 cells cultured with high glucose and low glucose were significantly enhanced,and the levels of m RNA and protein of TRIB3 in T24 and EJ cells were significantly increased,and those in T24 and EJ cells were significantly higher than those in T24 cells.After transfection with TRIB3 si RNA,the proliferation and invasion level of the cells were detected.The expression of p ERK1/2,ERK1/2,CCND1,MMP-9 were detected before and after TRIB3 interference,and the expression of p ERK1/2,ERK1/2 were significantly changed before and after interfering with TRIB3,The addition of MAPK pathway inhibitor U0126 significantly inhibited the proliferation and migration of T24 cells stimulated by TRIB3.Conclusion:1.By gene chip detection,it was found that the differentially expressed RNA in bladder cancer cells at high glucose concentration.They may be associated with high glucose-induced bladder cancer cell lines.The development of cancer is closely related to the process of migration and invasion,which provides a basis for further screening and progression of bladder cancer under high glucose.TRIB3 is selected as the research object by screening.2.The bladder cancer complicated with diabetes group had higher BMI,higher proportion of hypertension,lower hemoglobin,higher fasting blood glucose,higher tumor grade and stage,and higher expression of TRIB3 compared with the simple bladder cancer group.The tumor tissues of bladder cancer complicated with diabetes mellitus had high expression of TRIB3,stage,higher grade and higher fasting blood glucose at recurrence.3 HG overexpresses TRIB3 can promote the migration and proliferation of bladder cancer cells.TRIB3 promotes the proliferation and migration of bladder cancer cells through MAPK pathway. |
