Establishment Of Circulating Tumor Cell Sorting Method And Its Application In Prostate Cancer

Objective:Design and establishment a microfluidic chip enrichment platform and optimize the sorting efficiency,so that it can be used for the CTC isolation from peripheral blood specimens of prostate cancer,and to evaluate the application value of CTC in the diagnosis of prostate cancer.Methods:1.Design and manufacture a microfluidic chip,and test the chip by tumor cells standard models with gradient concentration to adjust the parameters of the microstructure and optimize the sorting efficiency of the chip.2.Use the immunomagnetic bead method to verify the sorting efficiency of the microfluidic chip for CTC,and use immunofluorescence technology and flow cytometry to identify the captured CTC.3.Carry out CTC sorting and biological identification on clinical samples,count the results of CTC sorting and identification,and analyze the correlation between CTC count levels and cancer stage or metastasis degree of prostate cancer patients.Results:1.Established a microfluidic chip enrichment platform,and the sorting purity is close to 100% for the tumor cells standard models,which is conducive to the subsequent analysis of tumor cells.2.The sorting rate of the density gradient tumor cell standard model can exceed80% at an appropriate density,and the identification results of the captured CTC by immunofluorescence technology and flow cytometry are consistent.3.The level of CTC in peripheral blood of patients with prostate cancer was significantly higher than that of patients with prostate hyperplasia.The number of CTC detections increased significantly with the increase of prostate cancer stage and prostate cancer metastasis degree.Conclusion:An effective CTC sorting method has been established,which can be used for the isolation and detection of CTC in clinical samples;the level of CTC count and the expression of its surface markers have important clinical application value in assessing the progression of prostate cancer.