ABC family proteins are often involved in multidrug resistance in cancer.34 Among the nine PTX-related ABCs, we found ABCB1 protein becoming the only one that was selectively induced by MUC1, although mRNA levels of ABCC1 and ABCC5 were also elevated by MUC1. (ABCB1). Remarkably, targeted suppression of EGFR or ABCB1 by both shRNAs and inhibitors efficiently reversed chemoresistance. Moreover, co-administration of the inhibitors of MUC1CEGFRCABCB1 with paclitaxel significantly blocked not only tumor growth but also relapse in xenograft mouse model. Our data collectively support a model in which MUC1 induces acquired chemotherapy resistance by upregulating ABCB1 in an EGFR-dependent manner, providing a novel molecular basis of using the EGFR inhibitor in MUC1-positive cancers to prevent chemotherapy resistance. Chemoresistance is one of the important mechanisms responsible for tumor recurrence and poor prognosis in a variety of tumor types.1, 2, 3 Paclitaxel (PTX) is a tubulin-disrupting drug in the management of a wide range of tumors.4, 5, 6 Although studies possess uncovered the mechanisms of PTX resistance in several malignancies, many LX-1031 critical issues remain, warranting further investigation. ATP-binding cassette (ABC) transporters are shown to selectively pump out cytotoxic medicines from cells resulting in multidrug resistance.7 The human being ABC transporter B1 (ABCB1), also known as p-glycoprotein (Pgp), is one of the well-characterized ABC transporters with the broadest substrate specificity. Many chemotherapy medicines for malignancy are substrates for ABCB1, including PTX, vincristine, doxorubicin and etoposide.8, 9 ABCB1 is found overexpressed in malignancy individuals with poor response to chemotherapy.10, 11, 12 To overcome ABCB1-induced chemoresistance, several pharmacological inhibitors have been developed but with limited success in clinic because of toxicities, which is primarily attributed to the critical functions of ABC transporters in various normal tissues in the physiological clearance of catabolites and xenobiotics.13, 14 Mucin 1 (MUC1) is a transmembrane glycoprotein. In normal tissues, MUC1 distributes around the apical surface of luminal epithelial cells and forms a mucinous gel with other mucin members to protect the underlying epithelia.15, 16 However, MUC1 is aberrantly glycosylated and overexpressed in many carcinomas and associated with poor outcomes,17, 18 including cervical cancer19 and lung cancer.20 Abundant evidence indicates oncogenic functions for MUC1, which (1) promotes receptor tyrosine kinases activation and downstream signaling21, 22 (2) attenuates the apoptotic response to Mouse monoclonal to KLHL25 genotoxic and oxidative stress23 and regulates the Wnt/were used as loading control for nuclear and cytoplasmic protein separately. (c) HeLa229 (P) and HeLa229/TR cells were treated with (TR/E) or without (TR) erlotinib (20?data, PTX treatment induced elevated expression levels of MUC1, ABCB1, and marked increase of EGFR nuclear localization in tumor tissues (Physique LX-1031 7c). Of notice were that these effects LX-1031 were only obvious in HeLa229/shCTL tumor but not in HeLa229/shMUC1 tumor (Physique 7c), supporting a MUC1 dependency. TUNEL staining revealed that PTX treatment induced more apoptosis in HeLa229/shMUC1 tumors than that in HeLa229/shCTL tumors (Physique 7d). To examine the contribution of the MUC1/EGFRCABCB1 axis to tumor chemoresistance, we treated the HeLa229/shCTL tumor-bearing mice with PTX in combination with verapamil or erlotinib. Similar to the sensitizing effect of shMUC1, verapamil or erlotinib substantially augmented PTX-induced inhibition of tumor growth (Physique 7e,Supplementary Figures S6A and S6B). Of notice was that there was little difference LX-1031 in body weights of mice within groups of drug alone and combination treatment (Supplementary Physique S6C), indicating that the treatments did not cause significant toxicity. These data collectively support a critical role of the MUC1/EGFRCABCB1 axis in acquired chemoresistance of HeLa229 cells, and moreover that targeting this axis can effectively overcome the chemoresistance. Open in a separate windows Physique 7 Co-administration of the inhibitors of MUC1CEGFRCABCB1 axis and PTX prevents tumor relapse. (a and b) Six-week-old female BALB/c nude mice were subcutaneously injected with 2.5 106 HeLa229/shCTL cells or HeLa229/shMUC1 cells in ventral flanks. When tumor reached approximately 4?mm 4?mm, the mice were injected intraperitoneally with PTX at 40?mg/kg every three days for 15 days. The tumor sizes were measured every 3 days following PTX treatment. The tumor volume was calculated according to the formula: V=length width2/2. The data indicated mean with S.E.M. of six mice in each LX-1031 group. (b) At the 45th day, all mice were killed and tumors were excised and photographed. (c and d) IHC stainings (c) or TUNEL assay (d) of tumor tissue sections were carried out. (e) 2.5 106 HeLa229/shCTL cells or HeLa229/shMUC1 cells were subcutaneously injected in ventral flanks of 6-week-old female BALB/c nude mice. When the tumor reached 4?mm 4?mm, the mice.