Preliminary Study Of Circulating Tumor Cells In Recurrent Glioma

Objective:To investigate the relationship between recurrent gliomas and circulating tumor cell(CTC)by detection of circulating tumor cells in peripheral blood of patients with recurrent gliomas by subtraction enrichment-immunofluorescence in situ hybridization(SE-iFISH).Methods:From July 2018 to June 2019,20 patients with recurrent glioma and 10 healthy volunteers were enrolled.We collected the information and pathological types of those patients with recurrent gliomas.Peripheral blood samples were enriched by differential phase enrichment for cells that may be circulating tumor cells in the blood samples.And the enriched cells were treated with chromosome 8 probes,and stained with DAPI,CD45,CD133.Treat cells with abnormal fold multiples of chromosome 8 and DAPI~+and CD45~-as CTC.SPSS 23 was used for statistical analysis.T test was used for measurement data conforming to normal distribution.The Mann Whitney U test was used for measurement data not conforming to normal distribution.Kruskal Wallis H test was used for comparison of statistical differences between different subgroups,with P value<0.05 as statistical significance.Results:In this study,between July 2018 and June 2019,20 patients with recurrent glioma and 10 healthy volunteers were reflected in the present study.CTCs were found in 95%(19/20)of 20 patients with recurrent gliomas.The detection range of CTC was 2-111/6ml blood(median 7.5/6ml blood).The total number of CTC detected was 378,among which 87 were triploid(87/378,23.02%),and 18 patients(18/20,90%).There were 55tetraploids(55/378,14.55%)from 15 patients(15/20,75%).There were 236(236/378,62.43%)super quintuples from 19 patients(19/20,95%).18 CD133~+CTC(18/378,4.76%)were detected from 6 patients(6/20,30%),and CD133~-expression was detected in the remaining 360 CTC(360/378,95.24%).The number of CTC was 62(62/378,16.40%)in the blood samples of who patients with grade II tumor,among which one was CD133~+CTC(1/378,0.26%).68 CTC(68/378,17.99%)were found in the blood samples of who patients with grade III tumors,of which 9(9/378,2.38%)were found in who There were 248 CTC(248/378,65.61%)in the blood samples of patients with grade IV tumors,among which 8 were CD133+CTC(8/378,2.12%);there was no significant difference in the number of CTC in patients with different grades of recurrent gliomas(P=0.558),no significant difference in the number of CD133~+CTC in patients with different grades of recurrent gliomas(P=0.418),triploid in patients with different grades of recurrent gliomas(P=0.207)There was no significant difference in the number of tetraploid(P=0.550)and super pentaploid(P=0.179).Two patients with CD133~+CTC detected in WHO gradeⅣtumor had an out of situ tumor recurrence.We should pay attention to the possible relationship between them.In the healthy volunteers group,only one haploid cell of chromosome 8 was found in one case,which was significantly smaller than that in the patients with recurrent glioma(P<0.05).Conclusions:Through this study,the results show that CTC exists in the peripheral blood of patients with recurrent gliomas.The SE-iFISH technology can be used to detect the sensitivity of CTC in the peripheral blood circulation of patients with recurrent gliomas;different levels and different pathologies have not been found.The relationship between the number of types of recurrent gliomas and the number of CTCs.Compared with low-grade recurrent gliomas,the number of tetraploid CTC and superpentaploid CTC has increased,but further proof is required;Patients with a large number of CTCs and the presence of CD133+CTC may be related to recurrence in situ.Due to the small sample size in this study,further studies with larger sample sizes are needed.