| ObjectiveBladder cancer is a common malignant tumor in the genitourinary system.It is clinically divided into non-muscle-invasive bladder cancer and muscle-invasive bladder cancer.Non-muscle invasive bladder cancer is prone to recurrence,which brings a substantial economic and psychological burden to patients.After infiltrating muscles,it is easy to invade and metastasize,endangering the patients’ life.The underlying mechanism of non-muscle-invasive bladder cancer progressing to muscle-invasive bladder cancer is currently unclear.There is increasing evidence that glycosylation is involved in the malignant progression of tumors.In addition,the traditional intravesical perfusion method for non-muscle-invasive bladder cancer after surgery has not been significantly improved for many years,and patient compliance is poor.Therefore,analyzing the malignant progress of non-muscle-invasive bladder cancer,revealing the key events,exploring the specific role and mechanism of glycosylation in the malignant progression of bladder cancer,simultaneous development of a new intravesical drug delivery system,is an essential issue to curb the malignant progress of bladder cancer and improve the overall survival rate of bladder cancer patients.MethodsIn this study,lectin staining and flow cytometry were used to characterize the N-glycosylation level of bladder cancer cell lines.Combined with flow sorting to select cells with high N-glycosylation level,through Transwell invasion assay,wound-healing assay,sphere formation assay,clone units formation assay,to explore the effect of N-glycosylation on the invasion,migration,and stemness of bladder cancer cells.Subsequently,immunohistochemistry was conducted to characterize the N-glycosylation status of the mouse bladder carcinoma in situ model.Through bioinformatics analysis,we discovered the key glycosylation gene ST3GAL5 in the malignant progression of bladder cancer(T1G3 to T2 transformation).We then revealed the specific role of ST3GAL5 in the malignant progression of bladder cancer through gain and loss of function experiments,and ST3GAL5 downstream pathway activator and inhibitors were utilized to explore the mechanism.For the development of a novel drug delivery system for intravesical perfusion,magnetic Iron oxide was complexed to the surface of yeast cells by tannic acid,while a metal-organic framework was then synthesized in situ on the surface to form a cell robot capable of being driven under an external magnetic field.The prepared cell robots were characterized by laser confocal microscopy,flow cytometry,scanning electron microscopy,energy dispersive X-ray spectroscopy,and dead-live staining,followed by studies of their motility under rotating magnetic fields and in vitro evaluation of the killing of bladder cancer cell lines by Adriamycin.ResultsThe experiment found that cells with different N-glycosylation levels in the cell lines cultured in the 2D state did not have significant differences in invasion,migration,and sternness.Still,at the 3D level,that is,in the mouse bladder cancer model,high N-glycosylation levels cells are indeed at the forefront of muscles,indicating their high invasion and migration ability.Next,bioinformatics analysis data showed that ST3GAL5 was significantly negatively correlated with malignant progression of bladder cancer.Overexpression of ST3GAL5 can inhibit the invasion and migration of bladder cancer cells,and interference with ST3GAL5 can promote the invasion and migration of bladder cancer cells.GSEA,cell line immunofluorescence,and tissue microarrays showed that ST3GAL5 was significantly positively correlated with ID2 expression.ID2 pathway activator and inhibitor experiments show that ST3GAL5 partially exerts its function by affecting the BMP4-Smad 1/5/8 pathway.The yeast-based magnetron cell robot was developed to provide excellent motility under the manipulation of an external rotating magnetic field and to maintain motion in different biological fluids.In vitro killing evaluation showed that the motile drug-delivered cell robot was significantly more effective in killing cells than the static drug delivery system.ConclusionIn summary,this study initially characterized the N-glycosylation status of different bladder cancer cell lines and studied its effect on the function of bladder cancer cells.The protective role of the glycosylation gene ST3GAL5 in the malignant progression of bladder cancer and part of the mechanism for exercising the protective function was determined,which provided new potential targets for the diagnosis and treatment of bladder cancer,and improved the network for glycosylation to regulate the malignant progression of tumors.The new cell-based motion drug delivery system accelerates the chance of drug binding to cells,improves treatment efficiency,shortens treatment time,has potential applications,and can improve patient compliance.Figure 38 Table 1 Reference 102... |
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