MiR-886-5p Inhibition Inhibits Growth and Induces Apoptosis of MCF7 Cells

Lei-Lei Zhang,Jiang Wu,Hong Jing,Qiang Liu,Zhu-Lei Sun,Yan Zhang

doi:10.7314/apjcp.2014.15.4.1511

Abstract

To explore the molecular mechanisms of miR-886-5p in breast cancer., we examined roles in inhibiting growth and migration of MCF-7 cells. MiR-886-5p mimics and inhibitors were used to express or inhibit MiR-886-5p, respectively, and MTT and clone formation assays were used to determine the survival and proliferation. Hoechst 33342/ PI double staining was applied to detect apoptosis. The expression of caspase-3, caspase-8, caspase-9, MT1-MMP, VEGF-C and VEGF-D was detected by Western blotting, and the levels of MMP2 and MMP9 secreted from MCF-7 cells were assessed by ELISA. MCF-7 cell migration was determined by wound healing and Transwell assays. We found that the growth of MCF-7 cells was inhibited upon decreasing miR-886-5p levels. Inhibiting miR-866-5p also significantly induced apoptosis and decreased the migratory capacity of these cells. The expression of VEGF-C, VEGF-D, MT1-MMP, MMP2, and MMP9 was also found to be decreased as compared to controls. Our data show that downregulation of miR-886-5p expression in MCF-7 cells could significantly inhibit cell growth and migration. This might imply that inhibiting miR-886-5p could be a therapeutic strategy in breast cancer.

Highlights

MicroRNAs are small noncoding RNAs that function as tumor suppressors or oncogenes by binding to the 3’-untranslated region (3’-UTR) of target mRNAs to modulate protein expression (Bartel, 2004; Zamore, 2005; Shyu et al, 2008)
The expression of caspase-3, caspase-8, caspase-9, MT1-MMP, VEGF-C and VEGF-D was detected by Western blotting, and the levels of MMP2 and MMP9 secreted from MCF-7 cells were assessed by ELISA
The results showed that the miR-886-5p inhibitor was able to induce apoptosis in MCF7 cells (Figure 2)

Summary

Introduction

MicroRNAs (miRNAs) are small noncoding RNAs that function as tumor suppressors or oncogenes by binding to the 3’-untranslated region (3’-UTR) of target mRNAs to modulate protein expression (Bartel, 2004; Zamore, 2005; Shyu et al, 2008). Bioinformatics and other in silico analyses have identified that an estimated one-third of all human genes are regulated by microRNAs (Lu, 2005; Volinia et al, 2006) These miRNA molecules participate in tumorigenesis as either oncogenes or tumor suppressors. To explore the molecular mechanisms of miR-886-5p in breast cancer., we examined roles in inhibiting growth and migration of MCF-7 cells. Results: We found that the growth of MCF-7 cells was inhibited upon decreasing miR-886-5p levels. Conclusions: Our data show that downregulation of miR-886-5p expression in MCF-7 cells could significantly inhibit cell growth and migration. This might imply that inhibiting miR-886-5p could be a therapeutic strategy in breast cancer

Methods

Results

Conclusion