Detection And Significance Of EpCAM And CK19 In Circulating Tumor Cells Of Non-Small Cell Lung Cancer

Objective To detect the mRNA level of EpCAM and CK19 in circulating tumor cells of patients with non-small cell lung cancer by real-time quantitative PCR,and to analyze the clinical value of the EpCAM and CK19 in the early diagnosis,curative effect monitoring and prognosis evaluation of NSCLC.Methods EpCAM and CK19 cDNA were obtained from non-small cell lung cancer cell line H2009 by RT-PCR method.EpCAM was linked with pc-DNA3.1vector and CK19 was linked with pGEM-T vector respectively to construct the two-target gene normalization plasmid as positive standard templates for fluorescence quantitative PCR detection.The successful positive standard templates were serially diluted at 10~7,10~6,10~5,10~4,10~3,10~2,and 10~1 copies/?l respectively to generate a standard curve for real-time PCR.From July 2016 to July 2017,100 patients diagnosed with non-small cell lung cancer,were enrolled in Department of Thoracic Surgery,Beijing Chest Hospital,Affiliated to Capital Medical University.All the patients met the enrollment criteria and planned to undergo surgical treatment.At the same time,40 patients with lung benign disease and 40 healthy volunteers were entrolled as control.Total RNA from PBMC was extracted and cDNA was obtained by reverse transcription.The mRNA of EpCAM and CK19 in NSCLC patients and control group were detected by probe fluorescence quantitative PCR.Results 1)EpCAM and CK19 in NSCL patients were significantly higher than control(P=0.00),and the difference was statistically significant;2)The mRNA level of EpCAM and CK19 were related to tumor stage.More EpCAM and CK19represented more worse of tumor.In stage I NSCLC,CK19 was significantly higher than EpCAM(P=0.002).3)in stage I squamous-NSCLC,CK19 was significantly higher than EpCAM(P=0.017);4)There was a correlation between CK19 and EpCAM in NSCLC patients.Conclusion 1)circulating tumor cells may be used as an important evaluation indicator for early diagnosis of patients with NSCLC.2)Detecting CTCs'CK19 and EpCAM by real-time quantitative PCR may become a simple and feasible method for early screening and diagnosis of NSCLC and the difference of CK19 and EpCAM can determine the pathological type of NSCLC,which provide a certain reference value for the choice of clinical treatment.