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miR-152 down-regulation is associated with MET up-regulation in leiomyosarcoma and undifferentiated pleomorphic sarcoma

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Abstract

Purpose

Highly aggressive adult soft tissue sarcomas (STS), i.e., leiomyosarcomas (LMS) and undifferentiated pleomorphic sarcomas (UPS), present complex genomic anomalies and overall 5-year survival rates of 20 to 40%. Here, we aimed to identify new biomarkers that may be employed to improve the treatment of non-translocation STS patients. We validated 12 miRNAs implicated in tumor development using primary STS samples and selected miR-152 for further analysis in STS-derived cell lines.

Methods

59 primary STS samples (27 LMS and 32 UPS) and 10 matched normal control tissues were included in the study, as well as 3 STS-derived cell lines (HT1080, SW872 and SKLMS1) and a normal control mesenchymal cell line (hMSC). miRNA expression analyses were performed using a TaqMan microRNA Array platform and qRT-PCR (miR-152), respectively. The expression levels of the putative miR-152 targets MET and KIT were assessed using qRT-PCR and immunohistochemistry on tissue microarrays, respectively. In addition, various functional analyses were performed before and after miR-152 transfection into SKLMS1 cells.

Results

We found that 12 pre-selected miRNAs were down-regulated in primary STS tumor samples compared to its normal control samples. A statistically significant miR-152 down-regulation was found to be accompanied by high MET and KIT mRNA levels in both the primary samples and the STS-derived cell lines tested. miR-152 transfection in SKLMS1 cells led to a reduction in KIT and MET mRNA and protein levels which, in turn, was associated with a transient down-regulation of the PI3K/AKT pathway, a transient decrease in cell growth, and a transient increase in both apoptotic and S-phase cells.

Conclusions

Our data indicate that over-expression of MET and KIT in primary STS samples and its derived cell lines is associated with miR-152 down-regulation. This shift may play a role in STS development and, thus, may be used to identify patients at risk. The effect of MET down-regulation on downstream signaling pathways, such as the PI3K/AKT pathway, may provide a basis for the future design of novel STS treatment strategies.

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Acknowledgments

The authors wish to thank Dr. Alba Balladelli for editing the paper and Ms. Cristina Ghinelli for her graphic work. This work was supported by 5‰ donation (Italy). Chiara Novello was supported by a FIRC Triennal fellowship “Mario e Valeria Rindi” (research project n°13748).

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Authors and Affiliations

  1. Laboratory of Experimental Oncology, Rizzoli Orthopaedic Institute, Via di Barbiano 1, 10 40136, Bologna, Italy

    Laura Pazzaglia, Chiara Novello, Amalia Conti, Serena Pollino, Piero Picci & Maria Serena Benassi

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  1. Laura Pazzaglia
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  2. Chiara Novello
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  3. Amalia Conti
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  4. Serena Pollino
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Correspondence to Laura Pazzaglia.

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All authors have read and approved the contents of this manuscript and declare that the work is original and has not been submitted or published elsewhere. The authors also state that there is no potential conflict of interest.

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The research protocol was approved by ethic committees of the Rizzoli Institute where the tumor samples were collected. All patients provided informed consent.

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Pazzaglia, L., Novello, C., Conti, A. et al. miR-152 down-regulation is associated with MET up-regulation in leiomyosarcoma and undifferentiated pleomorphic sarcoma. Cell Oncol. 40, 77–88 (2017). https://doi.org/10.1007/s13402-016-0306-4

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