Characteristics Of CD44~+ CD24~- ESA~+ Phenotype Cells Of Before And After Neoadjuvant Chemotherapy In Breast Cancer

Objective:The aim of this study is to isolate the CD44+CD24-ESA+phenotype cells in primary breast cancer cells and 1028 cell line and calculate the percentage;To identify the specificity of CD44+CD24-ESA+ maker to breast cancer cells by using the maker separate the subpopulation cells;To know whether neoadjuvant chemotherapy and chemotherapy in vitro have the ability to arise the percentage of CD44+CD24-ESA+ phenotype cells in breast cancer.Methods:1. Identify the CD44+CD24- subpopulation by double-staining immunohistochemistry technique. Calculate the percentage of CD44+CD24- subpopulation in breast cancer patients who have neoadjuvant chemotherapy and have no neoadjuvant chemotherapy respectively.2. The CD44+CD24-ESA+ subpopulation is separated from before and after neoadjuvant chemotherapy cells by flow cytometry.Identify the difference percentage between the before and after neoadjuvant chemotherapy,Then, Get the inter-relations between the CD44+CD24-ESA+ subpopulation and recurrence and distant metastasis in breast cancer.3. The EPI and PAC are added to human breast cancer cell line 1028.The CD44+CD24-ESA+ subpopulation was calculated from the dosing and control groups by flow cytometry.Identify the difference percentage between the two groups.Results:1. Cultured human primary breast cancer cells and established a human breast cancer cell line 1028 successfully.2. Identify the CD44+CD24- subpopulation by double-staining immunohistochemistry technique. Calculate the percentage of CD44+CD24- subpopulation in breast cancer patients who have neoadjuvant chemotherapy and have no neoadjuvant chemotherapy respectively. Discoveried that the percentage of CD44+CD24- cells in neoadjuvant chemotherapy group was significantly higher than the contral group.3. Compare the CD44+CD24-ESA+ subpopulation percentage between before and after neoadjuvant chemotherapy in the same patient by flow cytometry. Found that after neoadjuvant chemotherapy, CD44+CD24-ESA+ subpopulation(6.91±1.44)% was significantly higher than that before neoadjuvant chemotherapy(1.38±0.68)% in needle aspiration specimens. Testified neoadjuvant chemotherapy in breast cancer patients could enriche the percentage of the CD44+CD24-ESA+ subpopulation.4. The CD44+CD24-ESA+ subpopulation is calculated from the dosing and control groups in breast cancer cell line 1028.The percentage of CD44+CD24-ESA+ in adding EPI and PAC groups were (23±5.87)%,(25±6.38)%respectively. While the percentage of CD44+CD24-ESA+ in control group is (6±1.29)%.There was a significant difference(P<0.05).The chemotherapy in vitro can simulate the human body effect and enriche the percentage of the CD44+CD24-ESA+ subpopulation.Conclusions:1. Cultured human primary breast cancer cells successfully and established a human breast cancer cell line 1028 using PCM-2.2. Separate the CD44+CD24-ESA+ subpopulation using CD44+CD24-ESA+ marks by flow cytometry in primary breast cancer cells successfully.3. Identify the CD44+CD24- subpopulation by double-staining immunohistochemistry technique. Calculate the percentage of CD44+CD24- subpopulation in breast cancer patients who have neoadjuvant chemotherapy and have no neoadjuvant chemotherapy respectively. Discoveried that the new adjuvant chemotherapy group CD44+CD24-cells was significantly higher than the control group.4. Cultured human primary breast cancer cells in before and after neoadjuvant chemotherapy in the same patient.From this study,we can see neoadjuvant chemotherapy can kill the tumor cells and the tumor volume decreased.Meanwhile,the percentage of the CD44+CD24-ESA+ subpopulation was significantly increased. Testified that neoadjuvant chemotherapy in breast cancer patients could enriche the percentage of the CD44+CD24-ESA+ subpopulation. Neoadjuvant chemotherapy killed the cancer cells sensitive to chemotherapy, while leaving cancer cells resistant to chemotherapy. Get the inter-relations between the CD44+CD24-ESA+ subpopulation and recurrence and distant metastasis in breast cancer. In the perspective of stem cells to clarify the mechanisms of the development, recurrence and metastasis of breast cancer. And explainde the reasons for chemotherapy resistance in breast cancer patients, Then, provided clues to comprehensive treatment of breast cancer. 5. The EPI and PAC are added to human breast cancer cell line 1028.The CD44+CD24-ESA+ subpopulation is calculated from the dosing and control groups by flow cytometry. The chemotherapy in vitro can simulate the human body effect and enriche the percentage of the CD44+CD24-ESA+ subpopulation from multiple low-dose chemotherapy.