| Objective:This study investigated the relationship between chromosome ploidy of peripheral blood and pleural fluid neoplasm cells in advanced NSCLC and clinical features,chemotherapy efficacy,survival and distant metastasis of patients,providing a basis for accurate evaluation of NSCLC.Methods:58 patients with NSCLC with malignant pleural effusion confirmed by pathology from April 1,2014 to May 31,2017 in the Department of Respiratory,Subei People's Hospital,including 35 cases of adenocarcinoma and 23 cases of squamous cell carcinoma,AP regimen and TP regimen were used for 2 cycles respectively.The pleural effusion before treatment and the peripheral blood of some patients before and after treatment were collected.Tumor cells were enriched by Subtraction Enrichment(SE)and chromosome ploidy was identified and differentiated by immunofluorescence staining-immunosing-FISH(iFISH).To analyze the copy number characteristics of chromosome 8 in pleural fluid and peripheral blood tumor cells of patients with advanced NSCLC,and further study the correlation between chromosome ploidy and clinical features,therapeutic efficacy,survival and distant metastasis.Results:1.Tumor cells are classified into triploid,tetraploid and polyploid(copy number?5)based on chromosome 8 copy number.Before treatment,52 patients were detected by SE-iFISH in the pleural effusion of 58 patients with advanced NSCLC.The positive detection rate was 89.66%(52/58).Among the peripheral blood of 18 patients,16 were found to have circulating tumor cells(CTCs),and the positive detection rate was 88.89%(16/18).No significant correlation was found between pathological type,gender,age,smoking history,PS score,and tumor cell detection rate;2.After 2 cycles of standard chemotherapy,the number of aneuploid CTCs decreased from 7.75±6.74(range,0-29)to 4.19 ± 2.81(range 0-9),and the difference was statistically significant(P=0.030).Further analysis,after the end of 2 cycles of chemotherapy,7 of the 16 patients with positive CTCs were partially relieved(PR),6 were stable(SD),and 3 were disease progression(PD).The number of aneuploid CTCs in patients with PR after treatment was significantly decreased from 10.14±9.23(range,2-29)to 2.29±1.98(range,0-6),and the difference was statistically significant(P=0.048).In PD/SD patients,the number of aneuploid CTCs decreased slightly from 5.89 ±3.52(range 0-11)to 5.44 ± 2.65(range,2-9),and the difference was not statistically significant(P=0.77);3.Further analysis of CTCs chromosome 8 ploidy revealed that a total of 124 CTCs were detected in 16 of the 18 patients with NSCLC before treatment.According to the frequency of CTCs of different chromosome 8 copy numbers,the pre-treatment triploid,tetraploid and polyploid CTCs were calculated to be 56.25%,56.25%and 87.5%,respectively.Comparing the frequency of CTCs with different copy number of chromosome 8 before and after treatment,the results showed that the frequency of tetraploid and polyploid CTCs decreased after treatment,and the frequency of tetraploid CTCs decreased from 56.25%to 37.5%,respectively.The frequency of polyploid CTCs decreased from 87.5%to 62.5%,while the frequency of triploid CTC increased from 56.25%to 75.0%.According to the different chemotherapy effects of patients,patients were divided into three groups:the frequency of triploid CTCs in PR patients was from 42.86.%(before treatment)increased to 57.14%(after treatment);the frequency of tetraploid CTCs decreased from 57.14%(before treatment)to 28.57%(after treatment);the frequency of polyploid CTCs decreased from 100%(before treatment)to 42.86%(after treatment);the frequency of triple CTCs in SD patients increased from 66.67%(before treatment)to 83.33%(after treatment);the frequency of tetraploid CTCs decreased from 50.0%(before treatment)to 33.33%(after treatment);The frequency of ploidy CTCs decreased from 83.33%(before treatment)to 66.67%(after treatment);the frequency of triploid CTCs in PD patients increased from 66.67%(before treatment)to 100%(after treatment);the frequency of tetraploid CTCs did not occur.The change was 66.67%(before and after treatment);the frequency of polyploid CTCs was from 66.67%(treatment)Before treatment)increased to 100%(after treatment);4.The correlation analysis between the counts of aneuploid CTC and the prognosis of patients after treatment showed that there was no correlation between the triploid and tetraploid CTC counts and PFS and OS after treatment,P>0.05.Analysis of the relationship between polyploid CTC counts and PFS and OS after treatment showed that PFS and OS in patients with?2 polyploid CTCs after treatment were significantly shorter than those in<2 polyploid CTC patients[3(95%CI,2.542-3.458)months vs 4(95%CI,3.633-4.367)months,P=0.018;5(95%CI,2.228-7.772)months and 8(95%CI,5.783-10.217)months,P =0.002];5.According to the difference in chromosome 8 copy number of pleural effusion tumor cells,pleural fluid tumor cells and CTCs are divided into triploid tumor cells,tetraploid tumor cells and polyploid tumor cells.According to the proportion of atypical aneuploid tumor cells in 52 patients with positive pleural effusion cell detection,the patients were divided into three groups,one of which had a proportion of three and four tetraploids,50%,not included.The patients were mainly tetraploid,including 24 people with distant metastasis,19 with no distant metastasis,and 12 with tetraploid,including 2 with distant metastasis.There were 10 people without distant transfer;there were 15 people with polyploids,including 13 with distant metastasis and 2 with no distant metastasis.There were significant differences in distant metastasis between patients with triploid,tetraploid and polyploid(P<0.05).Further analysis showed that there was a statistically significant difference in distant metastasis between polyploid-based patients and non-polyploid patients,with P=0.000 and an OR of 26.250.Conclusions:1.The detection rates of pleural fluid and peripheral blood tumor cells in advanced non-small cell lung cancer by SE-iFISH system were 89.66%and 88.89%,respectively.No significant correlation was found between tumor cell detection rate and pathological type,gender,age,smoking history,and PS score,2.The change in the number of aneuploid CTCs is related to the therapeutic response,and monitoring the number of aneuploid CTCs has the potential to predict therapeutic effects.3.CTCs ploidy analysis is associated with chemosensitivity and drug resistance,with triploid CTCs showing persistent resistance to paclitaxel(PTX)or cisplatin(DDP)-based chemotherapy,tetraploid CTCs and polyploidy.CTCs may be associated with secondary resistance.Triploid CTCs counts may help predict the effects of chemotherapy.4.CTC ploidy in patients with advanced NSCLC has a certain correlation with survival.After 2 cycles of treatment,the PFS and OS of patients with?2 polyploid CTCs after treatment are lower than<2 polyploid patients,indicating more after treatment.Ploidy CTC predicts poor PFS and OS in patients with advanced NSCLC.5.Compared with patients with more triploid and tetraploid tumor cells,patients with higher proportion of polyploid tumor cells in pleural effusion before treatment of advanced non-small cell lung cancer are more likely to have distant metastasis. |
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