| ObjectiveBladder cancer is the most common malignancy of the genitourinary system at present.Its 5-year survival rate is high,but it is very easy to recur.Although regular cystoscopic examination,repeated bladder perfusion therapy and transurethral resection of bladder tumor(TURBT)can effectively control cancer cells,prevent recurrence of bladder cancer and improve the prognosis of patients,it will greatly increase the cost of treatment and seriously reduce the quality of life of patients,bringing severe burden to patients and their families.Current studies have suggested that epithelial-mesenchymal transformation(EMT)plays an important role in tumor progression and prognosis.However,studies on the regulatory genes and corresponding mechanisms in the EMT process of bladder cancer cells are still not perfect,and there are few reports on specific drugs to reverse EMT.Given single-celled sequencing technology in determining the unique advantages of molecular subgroup of bladder cancer cells,by sequencing single-celled looking for differentially expressed genes in bladder cancer,clarify its in bladder function and molecular mechanism in the process of EMT,for screening treatment,evaluation of treatment effect,improving the prognosis of patients with bladder cancer and improve survival rate has important significance.MethodsThe differentially expressed genes in the carcinoma and adjacent tissues of 4 patients with invasive bladder cancer were screened by single cell sequencing.These genes were further grouped by EMT score to screen out the key genes that play an important role in the EMT process of bladder cancer cells.Survival and prognosis were analyzed using TCGA database.Finally,it was determined that MTMR2 gene was not only differentially expressed in bladder cancer tissues,but also could regulate the EMT process and affect the prognosis of patients.Q-PCR was used to detect MTMR2 expression in bladder cancer cell lines such as SV,SW780,5637,MGH,UMUC-3,EJ and T24.Through the detection of MTMR2 in urinary exosomes of bladder cancer and normal people,the role of MTMR2 in the diagnosis of bladder cancer was analyzed.Immunofluorescence technique and immunohistochemical staining were performed in paraffin sections of bladder cancer to verify the expression level of MTMR2 at the protein level.The effects of MTMR2 gene on the movement and invasion ability of bladder cancer cells were verified by knockout and over expression of MTMR2 gene by using scratch healing experiment,matrix glue chamber migration and invasion experiment and other commonly used experimental methods in cell biology.Results372 differentially expressed genes in bladder cancer were identified by single-cell sequencing.After further grouping these genes by EMT score,45 genes were found to be closely related to the EMT process of bladder cancer.Survival and prognosis analysis using TCGA database showed that the high expression of MTMR2 gene could significantly shorten the disease-free survival of patients(HR 1.8,1.8P<0.05),and it could be used as an independent prognostic factor to evaluate the prognosis of bladder cancer patients.The expression of MTMR2 in urine and tissue samples of bladder cancer patients was significantly increased by q-PCR and immunohistochemical staining.The cell biology experiment also proved that the high expression of MTMR2 could significantly promote the migration,invasion and tumor stem cell characteristics of bladder cancer cells.ConclusionMTMR2 gene is not only differentially expressed in patients with bladder cancer,but also a key gene that regulates the EMT process of bladder cancer.It can be used as an independent prognostic factor to affect the movement and invasion of bladder cancer cells.The study of MTMR2 may provide new targets for individualized treatment of bladder cancer.Figure 7 Table 1 Reference 97... |
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