The Effect Of Adriamycin On Expression Of ABCG2、P-gp、ERCC1 In Human Triple-negative Breast Cancer Cells

Background and purpose Breast cancer is one of the most common malignant tumor in female in our country and because of its poor treatment, triple-negative breast cancer seriously threatens health of patients. Adriamycin is commonly used in clinical treatment of breast cancer. But some patients in clinical treatment or after the treatment occur recurrence or distant metastasis and it is easy to develop resistance to other chemotherapy drugs. How to overcome the resistance is the current difficulties in the treatment of triple-negative breast cancer. ATP-binding cassette transporters(ABC transporters) including P-glycoprotein,adenosine triphosphate binding cassette transporter protein superfamily member G2(ABCG2) are members of transmembrane proteins superfamily and are widely distributed throughout the body. The main function of ABC transporters is to utilize the energy of adenosine triphosphate(ATP) binding and hydrolysis to energize the translocation of various substrates(drugs, metabolites and other compounds) across membranes.Because they can help cancer cells protect themselves from cell growth inhibitors, ABC transporters play an important role in drug resistance and is the key to the success of chemotherapy.In order to investigate the mechanism of drug resistance in TNBC patients after treatment with adriamycin, this issue select human breast cancer cell line MDA-MB-231 as the object of our study.The MDA-MB-231 cells were exposed in proper concentration of adriamycin. The expression of ABCG2, P-gp and ERCC1 in cells which subsisted in proper concentration of adriamycin and the tolerance to other chemotherapy drugs was analyzed by a variety of methods.To preliminary analysis of the possible mechanism of breast cancer cells to adriamycin resistance.Method and Materials The MDA-MB-231 cells were exposed in proper concentration of adriamycin. The cells which subsisted in proper concentration of adriamycin was selected as the object of our study. The growth inhibition rate to cisplatin were analyzed by CCK-8 assay. The expression of ABCG2, P-gp and ERCC1 at protein levels in the MDA-MB-231 cells was detected by Western blotting.Results 1 The inhibition rate of adriamycin on MDA-MB-231 cells was on a dose-effect relationship. With increasing the concentration of adriamycin, the growth inhibition rate of MDA-MB-231 cells also increased. 50% inhibition concentration(IC50) of adriamycin for MDA-MB-231 cell line was 1.25μg/ml determined by CCK-8 assay. 1/2 IC50 of adriamycin was selected as follow-up experimental operation. 2 IC50 of cisplatin for cells which subsisted in proper concentration of adriamycin was analyzed by CCK-8 assay. The results showed that the drug-resistance of MDA-MB-231 cells to cisplatin increased as incubation time increasing. 3 Western blot assay displayed that the expression of ABCG2, P-gp and ERCC1 at protein levels in MDA-MB-231 cells exposed in adriamycin after 8-hour and 24-hour was significantly higher than that in non-interventional MDA-MB-231 cells. With increasing the intervention time of adriamycin, the expression of ABCG2, P-gp and ERCC1 at protein levels also increased.Conclusions 1. The expression of ABCG2, P-gp and ERCC1 at protein levels in MDA-MB-231 cells exposed in adriamycin was increased. 2. The expression of ERCC1 at protein levels in MDA-MB-231 cells which subsisted in proper concentration of adriamycin was increased and MDA-MB-231 cells had a certain drug resistance to cisplatin.