Investigation Of The Inhibitory Effects Of Antiparasite Drug Mebendazole To Chemoresistant Acute T Lymphocytic Leukemia Cells

Acute T-lymphocytic leukemia(T-ALL)is a type of highly invasive malignant hematologic disease,which is known for its poor prognosis.The exact pathogenetic mechanism of T-ALL is still unclear.Existing chemotherapeutic drugs generally have multiple side effects,and their effects can be affected by drug resistance.Studies have shown that more than 55%of T-ALL patients have Notchl activating mutations.Therefore,there is an emerging need to identify new drugs with low toxicity,which can overcome drug resistance of T-ALL.Previous studies indicated that antiparasitic drug mebendazole(MBZ)had inhibitory effects on a variety of tumor cells.In this work,we firstly discovered that MBZ could inhibit T-ALL cells growth and viability,which were measured by the MTS assay and Annexin V-FITC/PI flow cytometric staining.Then,fluorescence microscopic experiments identified that MBZ inhibited the polymerization of tubulins in T-ALL cells,which induced cell cycle arrest in the G2/M phase and activated the Caspase apoptosis pathway.RT q-PCR experiment provided evidence indicating that the expression of ABCB1 mRNA in CEM/C1 cells was 17 times higher than that in CCRF-CEM cells.We also found that the 24 h and 48 h MBZ treatments had different effects on the relative expression of P-glycoprotein(P-gp)mRNA in T-ALL cells.The results of Western Blot and RT q-PCR experiments showed that MBZ significantly knocked down the expressions of Notchl and its downstream transcription factors c-Myc and Hesl.To further validate the results obtained from the in vitro experiments,we established a murine model of T-ALL in immunodeficient nude mice with CEM/C1 cells.The BALB/c nu/nu mice were treated by intraperitoneal injection of MBZ(100 mg/kg).Peripheral blood was collected from the tail vein for Wright-Giemsa staining.T cell-specific surface marker CD4 was examined using flow cytometry.It was found that the T lymphocytes in the peripheral blood of the MBZ-administrated group were fewer than the control group and the expression of CD4 was decreased,indicating that growth of inoculated CEM/C1 cells was inhibited in vivo.The spleen and liver were removed for HE staining,which further proved that MBZ had therapeutic effects to the chemoresistant T-ALL models in vivo.Finally,Western Blot and RTq-PCR experiments proved that MBZ inhibited the expression of Notchl signaling pathway in vivo.Collectively,this study demonstrated that MBZ was a potent inhibitor to chemoresistant T cell acute lymphoblastic leukemia cells.