Expression And Function Study Of Long Non-Coding RNA SPRY4-IT1 In Bladder Cancers

Background Malignant bladder carcinoma are most common tumor in urinary system,, which threats to human health.Although the diagnosis and treatment of bladder cancer has been made great progress, but there are still many questions to solve;And its incidence is higher and higher year by year,the ratio of recurrence,metastasis and death still remains high, which cause heavy burden to the society,and bladder cancer is one of the most difficult curing cancer diseases. Surgery is suitable for early and metaphase bladder cancer patients, but it relapses easily;Patients with advanced bladder cancer who appear more pelvic lymph node metastasis and so on, could not surgery, with palliative care measures, such as radiation,chemotherapy and biological therapy in patients with high mortality.Therefore, further study on bladder cancer pathogenesis, to explore new therapeutic regimen has an important clinical significance, which especially for bladder cancer patients who are easy recurrence and not sensitive radiation and chemotherapy patients with adanced for palliative care has important significance.Long noncoding RNA’s transcription of this length is more than 200 nucleotides, scientists found that,although lnc RNA is no translation and non-encoding proteins,lnc RNA plays an important role in regulating in human tumor disease progress, closely associated with tumor growth progression and prognosis, which made a new sense for lnc RNA role in diseases such as tumor, completely abandoned the wrong mind in the past: Lnc RNA were transcriptional noise; and due to the number coding RNA molecules is much less than non-coding lnc RNA that the function is a wide range, the expression of lnc RNA are intrinsically related to human diseases, especially tumor disease that caused more and more attention of scientists.SPRY4 introns transcript 1(SPRY4- IT1)(gene ID: 100642175) is a kind of lnc RNA that is relative mature in the current research, and the function or mechanism need further study in tumor diseases.The previous literatures were reported that the abnormal expression of SPRY4- IT1 was closely related to breast, stomach, lung, kidney and brain tumors and other human tumors, which may be a kind of necessary disease factors that were associated with human cancer disease,and SPRY4- IT1 can be used as a new biological indicator of tumor disease detection,which still demands further validation in clinical practice.Objective To investigate SPRY4- IT1 relative expression and its significance in bladder cancer, and to explore SPRY4- IT1 function in normal epithelial cells and bladder cancer cells.Methods Collecting specimens of bladder cancer tissues and its matching in pairs of normal tissues adjacent to carcinoma specimens, to test the relative SPRY4-IT1 expression by real-time q RT-PCR in the specimens and cells of bladder cancer and to detect the correlation between the relative expression of SPRY4-IT1 and clinico-pathological characteristics.The specific small interfere RNA( si-SPRY4-IT1) was transfected into two bladder cancer cell lines respectively,and And overexpression plasmid pc DNA3.1- SPRY4- IT1 was transfected into normal bladder epithelial cells,and the transfection efficiency of si- SPRY4-IT1 and pc DNA3.1- SPRY4-IT1 was assessed by q RT-PCR.Then the CCK-8 assay, MTT assay,migration assay, enzyme-linked immunosorbnent assay( ELISA) and flow cytometry assay were used to detect proliferation, motility and apoptosis in bladder cancer celland normal bladder epithelial cells, etc.Results Compared with the bladder cancer tissue samples adjacent to carcinoma, the SPRY4-IT1 relative expression was obviously up-regulated in 68.33% of bladder cancer tissue specimens(P=0.026 3) and related to bladder cancer pathological classification(P<0.05) and TNM stage(P<0.001) of malignant bladder cancer,no relationship with age and sex etc. At the same time comparison to the normal bladder epithelium SV- HUC- 1, the SPRY4-IT1 relative expression was obviously increased in 5637(P=0.002 29) and T24(P=0.000 12) of two kind of bladder cancer cells. The relative expression of SPRY4-IT1 could be down-regulated by the specific small interference RNA si-SPRY4-IT1( P<0.001). the specific small interference RNA si-SPRY4-IT1 could remarkably inhibit the proliferation( P<0.001),suppress imigration(P<0.01) and also promote apoptosis(P<0.01). The expression of SPRY4-IT1 could be up-regulated by pc DNA3.1-SPRY4-IT1( P<0.001). Pc DNA3.1-SPRY4-IT1 could promote the proliferation( P<0.05), imigration(P<0.001) and inhibite apoptosis(P<0.01).Conclusion SPRY4-IT1 is up-regulated in bladder cancer tissues and cells, which is related with histologic grading, clinical TMN staging in malignant bladder tumor, can accelerate cell growth and participate the process of regulation in bladder cancer,etc.SPRY4-IT1 plays an oncogene role in promoting cell growth of bladder cancer and provide the direction for bladder cancer diagnosis and treatment, the next step of research is focused on how to translate it into clinical application.