miR-218 targets survivin and regulates resistance to chemotherapeutics in breast cancer

Abstract

Multidrug resistance (MDR) remains one of the most significant obstacles in breast cancer treatment, and this process often involves dysregulation of a great number of microRNAs (miRNAs). Some miRNAs are indicators of drug resistance and confer resistance to chemotherapeutic drugs, although our understanding of this complex process is still incomplete. We have used a combination of miRNA profiling and real-time PCR in two drug-resistant derivatives of MCF-7 and Cal51 cells. Experimental modulation of miR expression has been obtained by retroviral transfection. Taxol and doxorubicin IC50 values were obtained by short-term drug sensitivity assays. Apoptosis was determined by flow cytometry after annexin V staining, by caspase 3/7 and caspase 9 activity assays and the levels of apoptosis-related proteins bcl-2 and bax by real-time PCR and Western blot. miR target was studied using transient transfection of luciferase constructs with the 3 untranslated regions (UTR) of target mRNAs. Small interfering RNA-mediated genetic knock-down was performed in MDR cells and its modulatory effect on apoptosis examined. The effect of miRNA on tumorigenicity and tumor drug response was studied in mouse xenografts. miRNA profiling of two drug-resistant breast cancer cell models indicated that miR-218 was down-regulated in both MCF-7/A02 and CALDOX cells. Ectopic expression of miR-218 resensitized both drug-resistant cell lines to doxorubicin and taxol due to an increase in apoptosis. miR-218 binds survivin (BIRC5) mRNA 3-UTR and down-regulated reporter luciferase activity. Experimental down-regulation of survivin by RNA interference in drug-resistant cells did mimic the sensitization observed when miRNA-218 was up-regulated. In addition, resensitization to taxol was also observed in mouse tumor xenografts from cells over-expressing miR-218. miR-218 is involved in the development of MDR in breast cancer cells via targeting survivin and leading to evasion of apoptosis. Targeting miR-218 and survivin may thus provide a potential strategy for reversing drug resistance in breast cancer

MicroRNA-134 targets KRAS to suppress breast cancer cell proliferation, migration and invasion

“…Transfected cells were collected, counted and resuspended in single cell suspension (FBS-free culture medium). Subsequently, 1x10 5 cells were added to the upper chamber and 500 µl culture medium supplemented with 20% FBS, was added into the lower chamber as a chemoattractant. Following a 24 h incubation, any non-migrated cells were carefully removed from the top of the chamber using a cotton swab.…”

“…The etiology of breast cancer remains unclear, although genetic and epigenetic alterations are considered to contribute to tumorigenesis and progression (4). Despite advances in modern therapies for patients with breast cancer, including surgery, radiotherapy, hormonal therapy and various types of chemotherapeutic approaches using targeted and non-targeted drugs, numerous patients with breast cancer respond only transiently to conventional chemotherapy (5,6). A high proportion of these patients eventually exhibit tumor metastasis, which is a major contributor to cancer mortality (5,6).…”

“…Despite advances in modern therapies for patients with breast cancer, including surgery, radiotherapy, hormonal therapy and various types of chemotherapeutic approaches using targeted and non-targeted drugs, numerous patients with breast cancer respond only transiently to conventional chemotherapy (5,6). A high proportion of these patients eventually exhibit tumor metastasis, which is a major contributor to cancer mortality (5,6). Therefore, it is necessary to explore the molecular mechanisms underlying the tumorigenesis and progression of breast cancer in order to develop more effective treatments.…”

Knockdown of microRNA‑29a regulates the expression of apoptosis‑related genes in MCF‑7 breast carcinoma cells

“…Research has demonstrated that miR-218 is involved in breast cancer evasion of apoptosis by targeting survivin (26). This molecule has been targeted for therapy in cancer based on its selective expression in tumors, but not normal tissues (27,28).…”

miR-218 tissue expression level is associated with aggressive progression of gastric cancer

“…miRNA-218 (miR-218) is expressed in many cancers and plays critical roles in carcinogenesis, including in proliferation and apoptosis (Hu et al, 2015;Liu et al, 2015;Ran et al, 2015;Xie et al, 2015;Zhang et al, 2015b). Its function in gastric cancer has also been investigated.…”