Research Of SPHK1 Expression In Bladder Cancer And Prognosis Of This Disease

BackgroundBladder cancer is the most common type malignant tumor in the Department of Urology clinical work, with high morbidity and mortality. At present,mainly methodes to its therpay include surgery, radiotherapy and chemotherapy.But the effect is not ideal, recurrence and progress after treatment is common, and its prognosis is poor. Clinical data analysis shows that from 2007 to 2012, 44 large urology center in China, a total of 14260 cases of bladder cancer in patients with recurrence after more than 70% of the patients receiving treatment transfer diffusion, 30% of the recurrent tumor malignant degree increased earlier. As molecular targeted therapy in tumor treatment is more and more attention, look for bladder cancer molecular targets to slow its recurrence diffusion transfer is of great significance.With the deepening study of the lipid group, found that scabbard fat can play a role of regulation of signaling pathways. Such as ceramide(ceramide, Cer), sphingosine(sphingosine, Sph), they can regulate intracellular signaling pathways caused cell cycle arrest and promoting apoptosis; Sphingosine 1 phosphate(sphingosine 1 phosphate, S1P) can promote cell survival. In the cells and promote apoptosis of Cer/Sph and promote the survival of S1 P between dynamic balance, the balance determines the fate of the cells, its action is called "sheath" fat- rheostat(sphingolipid- rheostat). Sphingosine kinase(sphingosine kinases, Sph Ks) is one of the important enzymes, the balance will promote apoptosis of Cer/Sph S1 P converted to promote survival. Thus inhibition of sphingosine kinase 1(sphingosine kinase 1, Sph K1) activity can not only reduce the S1 P that promote survival, at the same time also can increase to promote apoptosis and periodic block Cer/Sph, can be used as a means for the treatment of tumor. This study make Sph K1 as a target.Explore SphK1 in its role in the development of bladder urothelial carcinoma and its mechanism.To provide theoretical basis for make Sph K1 as a molecular target to the diagnosis and treatment of bladder urothelial carcinoma.Materials and methods1. The cultivation of the cell level BIU-87 cell lines: for PMA SphK1 activator; DMS for SphK1 inhibitors; 0.9% NaCl reagent blank to bladder cancer cell lines in SphK1 activation BIU-87groups; SphK1 inhibiting group; Blank control group. With CCK 8 to measure each cell group’s proliferation.TranswellBoyden compartment model is used to test each BIU-87 cell group’s relative hitrate and mobility of change; Western-blot method is used to test each cell group’s Sph K1, ERK1/2 and p- ERK1/2 protein levels; With ELISA method to detect each cell culture supernatant of MMP- 2, MMP- 9, and the protein content of u PA and VEGF expression,Use qRT- PCR detection each groups of cell`s, SphK1,MMP-2 and MMP-9 and uPA mRNA level; Using Matrigel three-dimensional culture method to observe the cell ’s the vascular mimicry(vasculogenic mimicry, VM) forming ability.Results1.SphK1 activator can promote BIU- 87 cell invasion and migration, at the same time it can significantly enhance cell SphK1, ERK1/2 and p-ERK1/2 protein expression, and promote the MMP-2 and MMP- 9 and u PA protein secretion. Promote the formation of the three-dimensional cultivation of tubular VM. Significantly enhance VEGF protein expression. SphK1 inhibitors inhibit BIU- 87 cell invasion and migration, and inhibition of SphK1, ERK1/2 and p-ERK1/2 protein expression, and inhibition of MMP-2 and MMP-9 and u PA protein secretion; Three-dimensional training could not form in the tubular VM; Weakened obviously VEGF protein expression.2.All of the 37 pairs of bladder cancer tissues and adjacent normal tissue adjacent to carcinoma in measurement of SphK1 m RNA expression by q RT-PCR.The expression in bladder cancer tissues was significantly higher than in normal tissue adjacent to carcinoma express. In 153 cases of bladder cancer pathology biopsy Sph K1 protein expression in bladder cancer pathological stage(P=0.045), tumor pathological grading(P<0.001) significantly correlated. Through mapping Kaplan Meier survival curve showed that patients with high expression of SphK1 5-year survival rate was significantly decreased Objective(P<0.001). And it can be showed that high expression of SphK1 is an independent poor prognostic factors for this kind of disease by means of Multivariate Cox regression analysis.ConclusionSphK1 can promote bladder cancer proliferation, invasion, migration and inhibit the apoptosis of cell, inducing the formation of VM. Its mechanism may be through the activation of signal ERK1/2 to promote MMP- 2 and MMP- 9 and associated u PA protein secretion, further strengthen the baldder cancer cell migration ability and promote the expression of VEGF and play a role. We can make Sph K1 as a molecular target to the bladder urothelial carcinoma`s diagnosis and treatment.