Chemosensitivity Of Primary Breast Cancer Cells Before And After Neoadjuvant Chemotherapy With TAC Scheme

Objective:The aim of this study is to master the culture method of primary breast cancer cells in vitro; To find the feasibility to detect chemosensitivity by Collagen gel droplet culture drug-sensitivity test (CD-DST); To analyse the change of chemosensitivity to Docetaxel, Pirarubincin, Epirubicin, Navelbine and Cisplatin between before and after neoadjuvant chemotherapy with TAC scheme in primary breast cancer cells.Methods:1.291Patients who have no neoadjuvant chemotherapy but have an operation are defined as before neoadjuvant chemotherapy group and27patients who have it but not received pathologic complete remission with TAC scheme(DOC75mg/m2+THP40mg/m2+CTX600mg/m2, q3w) are defined as after neoadjuvant chemotherapy group. Two gruops of specimens are both collected form Januray2011to Januray2012. Primary breast cancer cells are obtained from two groups and cultured in vitro.2. Primary breast cancer cells are embedded in artificial extracellular matrix type I collagen gel and cultured by three-dimensional model which is similar to cells microenvironment in vivo. They are contacted suitable concentration (calculated based on the drug concentration curve) of chemotherapeutic drugs (Docetaxel, Epirubicin, Pirarubin, Navelbine, platin) and are stained. The accurate evaluation of drug sensitivity is made by Scion Image System. The result is a ratio between the Optical density value of group treated by drugs (T) and that of blank control group (C). By definition, primary breast cancer cells are sensitive to drugs when the ratio is less than or equal fifty percent. While the ratio is greater than fifty percent, they are defined as drug resistance. Single drug effciency is a ratio between the number of sensitivity and the total patients.3. All the data were analyzed by SPSS17.0statistical software. Count date between groups was compared by χ2test.P<0.05is defined as statistically significant difference. Results:1. Human primary breast cancer cells were cultured successfully and the growth relatively presented a stable state after two weeks from obtained from tumor tissue. Primary breast cancer cells showed shuttle adherent growth and were mixed with a small amount of interstitial cell which was polygonal extension growth.2. The CD-DST with the advantage of high overall predicting value and a comparatively small number of cells (3×103cells/drip), that expanded the range of application and made up for the inadequacy of other technologies. The CD-DST had the feasibility of clinical application.3. Before neoadjuvant chemotherapy with TAC scheme, the single drug effciency from high to low in the order is Docetaxel (50.9%), Pirarubincin (48.5%), Epirubicin (42.6%), Cisplatin (29.2%) and Navelbine (23.7%).The single drug effciency of docetaxel and anthracycline was obviously higher than that of cisplatin and navelbine. The single drug effciency between docetaxel and anthracycline have no significant difference (P>0.05). The single drug effciency between cisplatin and navelbine have no significant difference (P>0.05). In elderly breast cancer (age≥65years) group, the single drug effciency of docetaxel (χ2=9.202,P=0.002) is low. The low histopatholo-gical grade (grade Ⅰ-Ⅱ) and invasive lobular carcinoma were not sensitive to anthracycline(P<0.05).4. After neoadjuvant chemotherapy with TAC scheme, the single drug effciency from high to low in the order is Cisplatin (48.1%), Docetaxel (33.3%)=Navelbine (33.3%), Pirarubincin (22.2%)=Epirubicin (22.2%). The single drug effciency of cisplatin is obviously higher than that of pirarubin and epirubicin χ2=3.979, P=0.046). There is no significant difference between other groups (P>0.05).5. After neoadjuvant chemotherapy with TAC scheme, the single drug effciency of pirarubincin (χ2=3.979, P=0.046) and epirubicin (χ2.156,.P=0.041) were significantly reduced. While, that of cisplatin is higher (χ2=4.156, P=0.009). There is no significant difference in docetaxel and navelbine (P>0.05). Conclusions:1. The experiment successfully cultured human primary breast cancer cells obtained from fresh tissues of breast cancer.2. The CD-DST was a good chemosensitivity assay method and could detect sensitivity of drugs and guide clinical to individualy choose sensitive drugs.3. After neoadjuvant chemotherapy with TAC scheme, the single drug effciency of anthracycline was significantly reduced, this showed drug resistance. However, the single drug effciency of cisplatin is higher. For breast cancer after neoadjuvant chemotherapy with TAC scheme, the sequential chemotherapy is priority to cisplatin such as TP scheme and NP scheme.