Study On Bladder Cancer Cell Isolation And Molecular Analysis

Bladdr cancer is a kink of common malignant cancer in urinary system. Because of its important role in bladdr cancer suverliance, clinical non-invasive diagnosis methods have been widely investigated. In this work, in order to imporove the cell isolation effect and detection sensitivity, a cell capture-PCR-MCE method was developed. This developed method was valid for tumor cell isolation and flexible molecular characterization, possessing potential applications in biomedical researches. The contents of this research are as follows:(1)The methods for tumor cell isolation from body fluid were introduced. The history and new development of non-invasive bladder cancer diagnosis assays were reviewed. The significance, argument of the paper were expounded.(2) It was possible for non-invasive diagnosis of bladder cancer because tumor cells were exfoliated into urine when tumorigenesis. In this part, polycarbonate membrane and nickel microsieve were utilized to isolate tumor cells from urine samples with high efficiency. Moreover, cells isolated on the membrane were retrieved by a simple and fast dissolving procedure with dichloromethane. This filtration device achived high cell recovery rate over85%within20min, which was superior to traditional centrifugation method. Filter type, fixatives and flow rate for tumor cell isolation were investigated in this part. This developed filtration method was of low cost, high speed and environmentally friendly, and made it theoretically possible for clinical diagnosis.(3)Somatic mutations and hypermethylation of promoter CpG islands have been confirmed to be the main processes for the transformation of a normal cell into cancer and presented as promising biomarkers for BC detection. In this part, we combined somatic mutations (HRAS, FGFR3, TERT) and methylations (ALX4, RUNX3, SALL3and MT1A) into one panel for the development of multi-makers assay for non-invasive diagnostic and prognostic intervention. After PCR amplification, target fragments were separated by MCE with high-throughput and accuracy. This developed method was of low cost, high speed and sensitivity, which has potential application in early tumor diagnosis or prognosis.