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Cancer Cell International

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Open Access 01-12-2017 | Primary research

miR-335 negatively regulates osteosarcoma stem cell-like properties by targeting POU5F1

Authors: Xiaodong Guo, Ling Yu, Zhengpei Zhang, Guo Dai, Tian Gao, Weichun Guo

Published in: Cancer Cell International | Issue 1/2017

Abstract

Background

Evidence is accumulating to link cancer stem cells to the pathogenesis and progression of osteosarcoma. The aim of this study is to investigate the role of miR-335 in osteosarcoma stem cells.

Methods

Tumor spheroid culture and flow cytometry were applied to screen out osteosarcoma stem cells. Real-time quantitative PCR was used to detect the expression level of miR-335 in MG63, U2OS and 143B osteosarcoma stem cells. The relationship of miR-335 expression with osteosarcoma stem cells was then analyzed. Transwell assay and transplantation assay were performed to elucidate biological effects of miR-335 on cell invasion and vivo tumor formation. Western Blot and luciferase assays were executed to investigate the regulation of POU5F1 by miR-335.

Results

The expression of miR-335 in osteosarcoma stem cells was lower than their differentiated counterparts. Cells expressing miR-335 possessed decreased stem cell-like properties. Gain or loss of function assays were applied to find that miR-335 antagonist promoted stem cell-like properties as well as invasion. Luciferase report and transfection assay showed that POU5F1 was downregulated by miR-335. Pre-miR-335 resulted in tumor enhanced sensitivity to traditional chemotherapy, whereas anti-miR-335 promoted chemoresistance. Finally, the inhibitory effect of miR-335 on in vivo tumor formation showed that combination of pre-miR-335 with cisplatin further reduced the tumor size, and miR-335 brought down the sphere formation capacity induced by cisplatin.

Conclusions

The current study demonstrates that miR-335 negatively regulates osteosarcoma stem cell-like properties by targeting POU5F1, and miR-335 could target CSCs to synergize with traditional chemotherapeutic agents to overcome osteosarcoma.

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Title: miR-335 negatively regulates osteosarcoma stem cell-like properties by targeting POU5F1
Authors: Xiaodong Guo
Ling Yu
Zhengpei Zhang
Guo Dai
Tian Gao
Weichun Guo
Publication date: 01-12-2017
Publisher: BioMed Central
Published in: Cancer Cell International / Issue 1/2017
Electronic ISSN: 1475-2867
DOI: https://doi.org/10.1186/s12935-017-0398-6

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