| BackgroundExcept for cardiovascular and cerebrovascular diseases,tumor is the main cause of death,which can significantly affect the quality of life and increase the economic burden of patients.Each year,there are about 750,000 new patients with hepatocellular carcinoma(HCC)around the world and more than 600,000 patients died of the disease,which makes HCC as the second leading cause of cancer deaths worldwide.The number of new HCC patients in China has accounted for half of the global number,which makes HCC as the second major malignant tumors.Therefore,HCC has caused the joint concern of doctors and patients.Since the high recurrence rate after hepatectomy or TACE,the overall 5-year survival rate was low.Therefore,early detection and evaluation of the therapeutic effect and prognosis of HCC are particularly important.At present,traditional clinical pathological parameters such as imaging and pathology examination combined with tumor staging have been widely adopted to evaluate the progress and prognosis of HCC.However,when some tiny residual lesions or distant metastases can not be identified by imaging,these indicators would fail to evaluate the development of HCC.To some extent,these indicators would delay the diagnosis and treatment of HCC and they are not sensitive indicator.With the limited method of treatment and high recurrence rate and mortality,new testing methods are urgently needed to identify the response of therapy and evaluate the disease progression of HCC.Early in the formation and growth of a primary tumor,cells are released from the primary tumor and/or metastatic sites into the bloodstream.These tumor cells are called circulating tumor cells(CTCs).And CTCs was hardly detected in the peripheral blood of non-tumor patients.Since most CTCs would apoptosis or engulfed by macrophages,the presence of CTCs does not necessarily indicate the formation of metastatic lesions.And epithelial-mesenchymal transition(EMT)in CTCs refers to the adherent epithelial cells migrate to mesenchymal state.Aberrant activation of EMT has been implicated in the process of tumor metastasis,based on studies with human cancer cell lines and mouse models.CTCs can be separated in a non-invasive way,and can thus be used as a "liquid biopsy" to follow patients over time.These CTCs can provide vital information for a better understanding of tumor biology and tumor cell dissemination.Their molecular characterization offers the unique potential to better understand the biology of metastasis and resistance to established Therapies.PurposeWe adopted a new strategy to explore the relationship between the EMT process of CTCs and hepatocellular carcinoma(HCC).Furthermore,we intend to illustrate the potential diagnostic value of CTCs of distinct phenotypes in HCC.MethodsThe clinical data of 33 HCC patients and 10 healthy volunteers were collected retrospectively.By using the optimized CanPatrol CTC enrichment technique,patient blood samples of about 5 ml were collected,and CTCs were identified and characterized.The first step of this detection process was to isolate CTCs via a filter-based method;then,an RNA in situ hybridization(RNA-ISH)technique based on the branched DNA signal amplification technology was used to classify the CTCs according to EMT markers.The relationships between HCC CTCs and clinical characteristics were analyzed.Results(1)CTCs were detected in all 33 HCC patients.No CTCs were found in the blood of healthy volunteers.The mean age of the patients was 53.76(range,31 to 75)years,and the average tumor diameter was 7.63(range,1.7 to 17)cm.The average number of CTCs was 17.84(range,2 to 81),while the mean number of epithelial,epithelial-mesenchymal-mixed and mesenchymal CTCs was 0.82(range,0 to 5,4.59%),13.82(range,1 to 75,77.41%)and 3.21(range,0 to 21,17.99%),respectively(the details are shown in Tables 2 and 3).(2)The number of epithelial CTCs was related to tumor size(r = 0.456,p = 0.008),epithelial-mesenchymal mixed CTCs were related to tumor number(r = 0.421,p =0.015),and mesenchymal CTC was associated with metastasis and BCLC stage(r =0.375,p = 0.017;r=0.395,P=0.023).There was no significant correlation between CTC number and other clinicopathological factors,such as age,serum AFP level or cirrhosis.Conclusions(1)The optimized CanPatrol CTC enrichment technique has high sensibility.(2)Epithelial-mesenchymal-mixed CTCs seem to play an important role in EMT transition in HCC,mixed CTCs might be a vital factor for intrahepatic metastasis,and mesenchymal CTCs had the potential to be a predictor of extrahepatic metastasis. |
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