The Detection of Circulating Tumor Cells (CTCs) in Patients With Breast Cancer Undergoing Cryosurgery Combined With DC-CIK Treatment

Study Overview
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Brief Summary
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Circulating tumor cells (CTCs) have the potential to provide a surrogate for'real-time biopsy' of tumor biological activity. Enumeration and molecular characterization of CTCs in breast cancer could play an important role in diagnosis, predicting the risk for tumor recurrence, and providing novel target therapy biomarkers. In view of these facts, the investigators wanted to demonstrate the value of multiparameter flow cytometry in detecting human tumor cells of breast cancer in normal peripheral blood after cryosurgery with or without dendritic cell(DC)-cytokine-induced killers(CIK) treatment, and the investigators also compared the specificity with reverse transcriptase polymerase chain reaction (RT-PCR) method.

Detailed Description
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1 day before and 2 days after cryosurgery with or without DC-CIK treatment，approximately 17-mL ethylene diamine tetraacetic acid(EDTA)-blood was drawn by vein puncture from patients with breast cancer and healthy volunteers. The blood of the healthy volunteers will be used to evaluate the sensitivity and specificity and as negative control cells. To avoid contamination with skin cells, 2 mL blood will be discarded before the study samples will be taken.Briefly, the mononucleate cells will be separated from the blood over Ficoll- Paque for 20 min with 1800g at 4℃. The interface cells will be removed and washed, and the red blood cells(RBCs) will be removed using a lysis buffer followed by a repeated wash. The mononuclear cells will be counted and aliquot for RT-PCR and multiparameter flow cytometry on the basis of at least 2-3×106 cells for each methodology. The cell pellet will be resuspended in phosphate-buffered saline for multiparameter flow cytometry and in Trizol reagent for RT-PCR. Aim : Identification of CTCs may lead to better diagnosis and prognosis and could help to choose an adequate therapy.

Official Title
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The Detection of Circulating Tumor Cells (CTCs) in Patients With Breast Cancer Undergoing Cryosurgery Combined With DC-CIK Treatment

Conditions
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Neoplastic Cells, Circulating

Intervention / Treatment
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* Other: Flow cytometry (FCM)
* Other: RT-PCR


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Age:18-75 Karnofsky performance status >60 Diagnosis of breast cancer based on histology or the current accepted radiological measures. Classification tumor,nodes,metastasis-classification(TNM) stage: Ⅱ,Ⅲ,Ⅳ Will receive cryosurgery and/or DC-CIK treatment Life expectancy: Greater than 3 months Patients' routine blood test, liver function and kidney function have no obvious abnormalities Ability to understand the study protocol and a willingness to sign a written informed consent document Exclusion Criteria: Patients with other primary tumor except lung cancer History of coagulation disorders or anemia

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 75 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Control<br>Use Flow cytometry (FCM) and RT-PCR to test peripheral blood mononuclear cells(PBMCs) from healthy volunteer. | Other: Flow cytometry (FCM)<br>* Use FCM to test PBMCs/CTCs from volunteers/patients.<br>Other: RT-PCR<br>* Use RT-PCR to test PBMCs/CTCs from volunteers/patients.<br>|
| Cryosurgery group<br>Use Flow cytometry (FCM) and RT-PCR to test CTCs from patients who received cryosurgery only, 1 day before and 2 days after the cryosurgery. | Other: Flow cytometry (FCM)<br>* Use FCM to test PBMCs/CTCs from volunteers/patients.<br>Other: RT-PCR<br>* Use RT-PCR to test PBMCs/CTCs from volunteers/patients.<br>|
| DC-CIK treatment group<br>Use Flow cytometry (FCM) and RT-PCR to test CTCs from patients who received DC-CIK treatment only, 1 day before and 2 days after the DC-CIK treatment. | Other: Flow cytometry (FCM)<br>* Use FCM to test PBMCs/CTCs from volunteers/patients.<br>Other: RT-PCR<br>* Use RT-PCR to test PBMCs/CTCs from volunteers/patients.<br>|
| Cryosurgery with DC-CIK treatment group<br>Use Flow cytometry (FCM) and RT-PCR to test CTCs from patients received cryosurgery and DC-CIK treatment both, 1 day before and 2 days after the cryosurgery with DC-CIK treatment. | Other: Flow cytometry (FCM)<br>* Use FCM to test PBMCs/CTCs from volunteers/patients.<br>Other: RT-PCR<br>* Use RT-PCR to test PBMCs/CTCs from volunteers/patients.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| The number of Circulating Tumor Cells (CTCs) |  | Up to 6 months | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Breast Neoplasms, Circulating Tumor Cells