Application Of Glycolytic Marker HK2 For Detection Of Pleural Effusion

ObjectiveThe cytoglycolytic marker HK2 was used to label highly metabolically active cells in pleural effusion to clarify its value in the identification of malignant tumor cells in pleural effusion.To compare the advantages and disadvantages of the identification of hypermetabolic tumor cells based on HK2 and the identification of malignant pleural effusion by traditional cytopathological methods,and to reveal the significance of the combined application of HK2 marker and cytology in improving the sensitivity and specificity of cytological diagnosis of serosal effusion.Methods1.The data of 81 patients and 83 pleural effusion samples form Jun 2020 to Feb 2021 were collected in Zhejiang Cancer Hospital.Each specimen was diagnosed by histopathology or exfoliative cytology.Morphological characteristics and immunocytochemical stain of each specimen were analyzed and compared under the microscope.The benign and malignant pleural effusion and the primary source of malignant pleural effusion were determined by comprehensive diagnosis based on liquid-based cytology,cell block and immunocytochemical staining.2.The data of 79 patients and 81 pleural effusion samples form Jun 2020 to Feb 2021 were collected in Zhejiang Cancer Hospital.Each specimen was diagnosed by histopathology or exfoliative cytology.At the same time,EpCAM/CK/CD45/DAPI was used to label the suspected target cells in the pleural effusion,and then the glycolytic marker HK2 was used to confirm the target cells.All the cells can be rapidly imaged and analyzed with multiple channels with a scanning fluorescence microscope,and suspected tumor cells with high expression of HK2 were identified.3.The suspected tumor cells in part of the samples labeled with HK2 were taken out by microoperation for single-cell whole genome amplification and second-generation sequencing,and their copy number variation(CNV)was analyzed.4.Comparison of the advantages and disadvantages of identifying hypermetabolic tumor cells based on HK2 markers and traditional cytopathological methods for malignant pleural effusion in 81 samples.Results1.83 cytological samples were positive(46.99%),negative(28.92%),and atypical(24.09%).There was no statistical difference between the positive detection rate and the primary tumor and tumor type(p>0.05).2.In 81 cases of high metabolic tumor cells detected by HK2,the positive rate of HK2 diagnosis was 96.30%,and the cytological sample was 48.15%,indicating a significant statistical difference between the two groups(p<0.01).The sensitivity of tumor cells with high expression of HK2 was much higher than that of cytology,especially in lung cancer and digestive system tumors(p<0.01).There were also significant differences in the diagnosis of adenocarcinoma and squamous cell carcinoma(p<0.01).3.Four samples from different sources were randomly selected,of which 1 case was cytologically positive,2 cases were cytologically negative,and the other cases were cytologically atypical,all of which were positive after the diagnosis of HK2.CNV confirmed that the suspected tumor cells in the 4 samples had obvious and consistent copy number variation compared with the primary tumor,suggesting the high specificity of HK2 detection.Conclusion1.Traditional cytological diagnosis of malignant tumor cells while has high specificity,but the sensitivity is relatively low,which is the difficult simple morphology to determine types and sources of the organization,and easily affected by treatment.Combined with cell block and immunocytochemistry staining,although improve the organization types and sources of diagnostic rate to some extent,the increase sensitivity is relatively limited.2.The sensitivity of tumor cells with high expression of HK2 was significantly higher than that of cytology,but the source and type of tumor could not be further defined.3.The detection of tumor cells with high expression of HK2 combined with liquid-based cytology,cell block morphology and immunocytochemistry for tissue localization can improve the sensitivity.At the same time,it can also identify the type and origin of tumor cells and tissues,which can make good for deficiency.