| Background: Breast cancer is one of the most frequently diagnosed cancer and the leading cause of cancer death among women worldwide. Despite conventional therapies(surgery, chemo-radiotherapy and hormonal therapy) have improved the survival rates in breast cancer patients, the chances of recurrence and metastasis are inevitable. Cancer stem like-cells(CS-like cell, also known as stemloids) attract much more attention owing to their contribution to therapy resistance, cancer metastasis and relapse in breast cancer, which makes it a potential target for novel cancer therapeutic strategy. Flubendazole is widely used as a safe and efficacious anthelmintic drug for gastrointestinal parasites control in human, rodents and ruminants. Flubendazole, a member of benzimidazole families, has the typical benzimidazole moiety but with an added fluorine atom as the major structural, which makes it different from other benzimidazoles. Recent studies reported that flubendazole suppresses cell proliferation in vitro, delays tumor formation in xenograft models and displays preclinical activity by inhibiting tubulin polymerization in leukemia and lymphoma. In addition, flubendazole increased efficiency of traditional chemotherapy drugs for intestinal cancer. Moreover, CS-like cell related traits are contributing to the cancer cell proliferation and drug resistance. Therefore, we demonstrated a novel effect of flubendazole target breast CS-like cells.Objective: 1: We sought to explore whether flubendazole can inhibit the proliferation of breast cancer cells. 2: To explore whether flubendazole target CS-like cell. 3: To explore whether flubendazole inhibit cancer stem cell properties, such as promoting the differentiation, inhibiting migration and invasion of breast cancer cells. In conclusion, our studies want to uncovered a remarkable effect of flubendazole on suppressing breast CS-like cells.Methods: 1. We confirmed the cytotoxic effect of flubendazole to breast cancer cell lines MDA-MB-231, BT-549, MCF-7 and SK-BR-3 by cell counting assay and MTT assay. 2.To evaluated whether flubendazole inhibited tumorigenicity of breast cancer cell line MDA-MB-231 in vivo by using a xenograft tumor model. 3.We analyze the proportion of CS-like cell subpopulation in MDA-MB-231 and epi-MCF-7 cells based on the expression of CD44 and CD24(CD44high/CD24 low configuration) the expression after incubated with flubendazole by flow cytometry analysis. 4. We subsequently tested whether flubendazole inhibited anchorage-independent sphere formation. MDA-MB-231 and BT-549 cells were exposed to indicated doses of flubendazole(0, 0.125 and 0.25 μM) for 7 days. 5. We tested the expression of self renewal genes such as c-myc, oct4, sox2, nanog and cyclinD1 in MDA-MB-231 cells by western-blot assay. 6. To explore whether flubendazole induces breast cancer cell differentiation, we performed Oil Red O staining in cancer stem-like cell enriched MDA-MB-231 cells before and after flubendazole treatment(0.125 μM, 3 weeks). 7. Western-blot was used to check the differentiation markers expression.Results: We demonstrated a novel effect of flubendazole on breast cancer stem-like cells. Flubendazole inhibited breast cancer cells proliferation at both dose- and time-dependent manners and delayed tumor growth in xenograft models by intraperitoneal injection. Importantly, flubendazole reduced CD44high/CD24 low subpopulation and suppressed the formation of mammosphere and the expression of self-renewal related genes including c-myc, oct4, sox2, nanog and cyclinD1. Moreover, we found that flubendazole induced cell differentiation and inhibited cell migration. Consistently, flubendazole reduced mesenchymal markers(β-catenin, N-cadherin and Vimentin) expression and induced epithelial and differentiation marker(Keratin 18) expression in breast cancer cells.Conclusion: In conclusion, our findings uncovered a remarkable effect of flubendazole on suppressing breast cancer stem-like cells, indicating a novel utilization of flubendazole in breast cancer therapy. |
PDF Full Text Request