| Background:Tumor metastasis is the leading cause of death in cancer patients.Recent studies have shown that when the tumor is at the stage of precancerous lesions,tumor cells have invaded into the circulatory system,some of which can form micrometastasis.Therefore,the early diagnosis of cancer and early acceptance of effective treatment can significantly improve the prognosis of patients,which also can extend the survival of patients.The current study suggests that the key factor of tumor recurrence is the potential of tumor cell invasion.Circulating tumor cells(CTCs)are related to the process of tumor metastasis,the relevant process is called epithelial mesenchymal transition,which is the corresponding changes in tumor epithelial cells.It can decrease the binding power between the tumor cells,which enhanced the tumor migration and infiltration,which promote cells to fell off from the primary disease and the development of a distant metastasis.CTCs carry the information of primary tumor,CTCs detection can be used as "liquid biopsy",and becacuse of the convenience of the inspection method,microtrauma and safety,CTCs can reflect the biological characteristics of the primary tumor,which also plays an important role in the tumor diagnosis and treatment process.CTCs detection as a "liquid biopsy" in the early stage of the tumor can guide the adjuvant therapy,so as to achieve the purpose of individual tumor therapy.CTCs detection is of related to distant metastasis or micrometastasis when detected,so CTCs detection can be used as early detection of the tumor.At the same time,CTCs can also provide the relevant biological information of the tumors,monitoring detection CTCs changes of patients,which can prolong the survival time and improve the quality of life for patients.Related studies have found that CTCs detection in non-small cell lung cancer,breast cancer,colorectal cancer,kidney cancer and other malignant tumors have a good application,which can effectively assess the prognosis of the disease.Objectives:In this paper,we establish an effective method for the detection of circulating tumor cells.The method is combined with density gradient centrifugation and immunomagnetic beads method,we apply it in clinical trials,which can dynamic monitor the CTCs changes of advanced hepatocellular carcinoma(HCC)and colorectal cancer liver metastasis(CRLM)We use CTCs to evaluate the efficacy of cryoablation and survival analysis.Methods:1.A serial dilutions of different cell lines were spiked into healthy human blood samples to simulate the blood samples of caner patients.The blood samples were enriched and the recovery of the cells was calculated to verify the efficacy of the method.2.The dynamic changes of peripheral blood CTCs in 47 patients with advanced HCC and 55 patients with CRLM were analyzed by flow cytometry and real-time quantitative PCR.The survival of patients was also analyzed.Results:1.For different cell lines,the number of detected CTCs increased with the increased tumor cell concentration,the best correlation between the hepatocellular carcinoma cell lines and colorectal cancer cell lines are HepG2 and CX-1,which are of great significant correlation.2.Varying numbers of HepG2,CX-1 cells were spiking into peripheral blood of healthy volunteers to verify the recovery rate of CTCs in this method,and the recovery rate was significantly correlated with the numbers of spiking cells.3.The number of CTCs was significantly decreased at 7 days and 30 days after cryoablation in 47 patients with advanced HCC(P <0.01).4.The Ct valuce of CTCs markers increased after cryoablation in 47 patients with advanced HCC.2-Ct values were less than 1,indicating that the expression of tumor marker reduced after cryotherapy.Statistical analysis showed significant statistical significance(P <0.01)5.Kaplan-Meier survival analysis in 47 patients with advanced HCC underwent cryoablation showed that the survival rate of patients with CTCs increased significantly shorter than that of CTCs decreased(p <0.05).6.The number of CTCs was significantly decreased at 7 days and 30 days after cryoablation in 55 patients with CRLM(P <0.01).7.The Ct valuce of CTCs markers increased after cryoablation in 55 patients with CRLM.2-Ct values were less than 1,indicating that the expression of tumor marker reduced after cryotherapy.Statistical analysis showed significant statistical significance(P <0.01)8.Kaplan-Meier survival analysis in 55 patients with CRLM underwent cryoablation showed that the survival rate of patients with CTCs increased significantly shorter than that of CTCs decreased(p <0.05).Conclusions:1.CTCs enrichment assay is an effective and sensitive method for detecting CTCs from peripheral blood of tumor patients.2.The number of CTCs detection after cryosurgery decreased significantly,and the expression level of related tumor mRNA decreased significantly3.Patients with decreased numbers of CTCs have longer survival than those with an increased numbers of CTCs... |
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