| Objective:To master the Flow Cytometry of Cytokeratin 19(CK19).We have chosen the popular and cheap method of lymphocyte separation medium and Flow Cytometry to detect the CK19.After detection we analyse the leval of CK19 in peripheral blood in patients with RCC. According to the leval of CK19,we evaluate the clinical significance of CK19 in the peripheral biood among renal cell carcinoma.Methods:First of all we have chosen 32 healthy donors as control group.At the same time we have chosen 51 patients of renal cell carcinoma. We have founded a method of lymphocyte separation medium to enrich and then to detect CD45-CK19+ by Flow Cytometry between control group and test group. Second we have analysed the leval of CD45-CK19+ among test group as recording to different ages,genders,grades,stagings and so on.we have follwed with these patients to get the information of basical siutuation,location of metastastis.All patients of mRCC received the targert thrapy of sorafenib.Finally we have follwed up these mRCC 3 to 12 months.We have measured the diamiters of all metastasis before and after treatment.By the guidline of Recist of solid tumors we have evaluate the therapeutic effect,like CR,PR,SD,PD.Furthermore we have detected the leval of CK19 after 6 monthes since receiving targeted therapy.The results were analysed refer to the outcomes of international expriments about CD45-CK19+.The mumber of CD45-CK19+ in control group is below 60 per 100 thousands. If number of CK19 is above that leval,it is called positive.The marker of CTC were detected from the peripheral blood samples of 51 patients with RC,32 cases of healthy control. the blood samples of RCC were also collected after treatment.At last we use SPSS 13.0 to analyse the level of CK19 of different clinical stagings and different gradings.Results:5 cases of 32 in control group are positive,the rate of CK19 is 15.63%.35 cases of 51 in test group are positive,the rate of CK19 is 68.63%.The differences of two groups have stastical meaning(P<0.01).There are 24 cases of Ti,10cases of T2,8 cases of T3,9 cases of T4 with renal cell carcinoma.The positive rate of CK19 in different stages are 54.17%(13/24),60%(6/10),75%(7/8),100%(9/9). There were stastical meaning between different stages (P<0.05), The differences of T1-T2 and T3-T4 also have stastical meaning(P<0.05).If diveded by histodifferentiation there are 33 cases of G1,and 10 cases of G2 and 8 cases of G3,the rate of CK19 are 63.63%(21/33),70%(7/10) and 85%(7/8). There were stastical meaning between different groups (P<0.05), The differences of G1 and G2-3 also have stastical meaning(P<0.05). The rate of mRCC among CK19+ were high than that of CK19-(P<0.05).We have followed up 12 mRCC.The number cases of mRCC from positive and negetive CK19 are 9 and 3.At the end of expriment the number of CR,PR,SD,PD are 1,2,5,4.There is no diffrence between CK19 and effciency of target therapy among mRCC(P>0.05).Conclusion:We have tried to detect the expreesion of CK19 by Flow Cytometry.The trial has proved that this way can tell expression of CK19 form peripheral blood,and has some clinical meaning.The results have showed that the expression of CK19 is different among two groups,the rate of positive in low in healthy group, while high in RCC. Futhermore in advaced RCC is more high. By this exsperiment it is proved that the leval of CK19 in peripheral blood can be told,This way can forecast the prognosis of RCC. To some extent the prognosis of mRCC has correlate with CK19 theoretically, but no statistical meaning was foud between the two factors. In a word the leval of CK19 by FCM can disguise the leval of CTCs in peripheral blood.This method is worth to study... |
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