The Detection And Analysis Of Like-circulating Tumor Stem Cells From The Peripheral Blood Of Patients With Nasopharyngeal Carcinoma

Objecitves: Detect the content of cancer stem cells before and after the treatment of thepatients with nasopharyngeal carcinoma,and analysis of their expression levels in patientswith clinical stage, eiffcacy and prognosis, and then analysis its relationship with tumorrecurrence and metastasis.Methods: Extract10ml peripheral blood from healthy volunteers and our hospitalpatients with nasopharyngeal carcinoma for ifrst diagnosed （before and after treat-ment），and then extracted their peripheral blood mononuclear cells. Use the anti-CD45monoclonal antibody, anti-Epcam monoclonal antibody,anti-CK monoclonal anti?body,anti-CD44monoclonal antibody and anti-CD133monoclonal antibody to labelthe sample. CTC is the CD45-/Epcam+/CK+cells,CTSC is CD45-/Epcam+/CD44+orCD45-/Epcam+/CD133+cells.Flow cytometry detection of CTC and CTSC’s content,and analysis the correlation of expression levels before and after treatment in pati?ents with clinical staging.Results:1.Only one case of10healthy volunteers have CTC；CTSC were not detected inthe healthy population.2.Beforetreatment the CTC detection rate is72%（36/50cases）, CD45-/Epcam+/CD44+is68%（34/50cases）, the CD45-/Epcam+/CD133+is40%（20/50cases）; CTC detection rate after treatment decreased to60%（30/50cases）,CD45-/Epcam+/CD44+reduced to58%（29/50cases）,CD45-/Epcam+/CD133+reduced to30%（15/50cases）.3.Beforetreatment,No-i staging of NPC patients:the rate of CTC <5is90.9%（20/22cases）,is9.1%（2/22cases）; N_2-3 staging, the rate of CTC <5and is thesame （14/28cases）, the difference is statistically signiifcant （P=0.002）. N_0-1staging,CD45-/Epcam+/CD44+cells detected in82.1%（23/28cases） patients and do notdetect in17.9%（5/28）. N_2-3 staging,CD45-/Epcam+/CD44+cells from patients with donot detect CD45-/Epcam+/CD44+cells in patients is each of50%（11/22patients）; distribution has a signiifcant difference （P=0.016）. This indicates that the number ofpatients with CTC and CD45-/Epcam+/CD44+cells for N staging have a positivecorrelation.4.After treatment, the staging of T1-2： CTC <5accounted for92.0%（23/25cases），^5accounted for8%（2/25cases）; T_3-4 staging,＜5accounted for56%（14/25cases）, accounted for44%（11/25cases）; and the difference has statistically signiifcant （P=0.004）. Similarly, in the N staging has the same results as theT stage, the difference is also statistically signiifcant （P=0.002）. For T stageand N stage,CD45-/Epcam+/CD44+cell number showed the same rule with the CTC，that is, with T and N staging increases, CD45-/Epcam+/CD44+cells detectableamount is increased, and the difference has statistically signiifcant （P=0.045，T staging; P=0.002，N stage）.5.Patients before and after treatment in a variety of cell number display t-hree trends: increasing, reducing and unchanged,in which the largest part is reduc?ing，but no correlation with the clinical indicators.Conclusions:1. In the peripheral blood of patients with nasopharyngeal carcinomat-here are circulating tumor cells and cancer stem cells,and use the lfow cytometrycan detect and analysis them.2.Before treatment, the patient’s CTC and CD45-/Epcam+/CD44+cells’ level and N staging have correlation.3.After treatment, the patient’s CTC and CD45-/Epcam+/CD44+cells’ level and T、N staging have correlation.4.CD133as nasopharyngeal cancer stem cell markers in the peripheral blood of patients can be detected,but the detection rate is lower than the CD45-/Epcam+/CD44+cells before and after treatment, and do not have a correlation with the patient’s clinical indicators.