Molecular Neurobiology

, Volume 54, Issue 10, pp 8162–8169 | Cite as

SIRP Alpha Protein Downregulates in Human Astrocytoma: Presumptive Involvement of Hsa-miR-520d-5p and Hsa-miR-520d-3p

  • Ravindra Pramod Deshpande
  • Y.B.V.K Chandra Sekhar
  • Manas Panigrahi
  • Phanithi Prakash Babu
Article
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Abstract

Astrocytomas are the most common brain tumors with poor survival in malignant forms. Signal regulatory protein alpha (SIRP alpha) is a transmembrane protein expressed on immune cells and macrophages and is reported to modulate tumor cell phagocytosis. In the present study, we investigated the involvement of miR-520d-5p and miR-520d-3p in regulation of SIRP alpha expression. Here, we report mRNA and protein expression profile of SIRP alpha in 39 surgically resected human astrocytoma tissue samples and 14 control brain tissue samples. Transcript expression pattern was studied by real-time PCR while Western blotting and immunohistochemistry were used to evaluate protein expression. Expression profile of miR-520d-5p and miR-520d-3p was studied by real-time PCR. Computational prediction was employed to analyze the binding of miR-520d-5p and miR-520d-3p for SIRP alpha mRNA. It is evident from preliminary investigation that SIRP alpha transcripts are expressed in control brain tissues, increased in low-grade (grade II) tumor tissues, and decreased with further grade progression (P < 0.05). SIRP alpha protein was moderately expressed in control brain tissues but under-expressed in low- and high-grade tissue samples (P < 0.05). Immunohistochemistry results further confirmed Western blot outcomes. Computational prediction supplemented with 3′ and 5′UTR targeting analysis and correlation studies reveals that hsa-miR-520d-5p (P = 0.028, R 2 = 0.94) (95 % CI 0.15 to 0.99) and hsa-miR-520d-3p (P = 0.027, R 2 = 0.94) (95% CI 0.17 to 0.99) may be the putative microRNAs involved in regulation of SIRP alpha protein expression. Real-time PCR expression profile depicts that mature form of both miRNAs is significantly overexpressed in low-grade (GII) tumor tissue samples compared to control and high-grade (GIII and GIV) tissue samples. MiR-520d-5p and miR-520d-3p were found with expression pattern similar to SIRP alpha transcripts. We show that SIRP alpha protein is under-expressed in low and high grades of astrocytoma patients’ tissue samples. Control brain tissues were found to be positive with SIRP alpha protein expression. Real-time PCR expression analysis confirms that miR-520d-5p and miR-520d-3p expression levels were significantly correlated with SIRP alpha transcripts in control, low-grade, and high-grade tissue samples. Computational prediction further evidenced for binding sites of these miRNAs on 3′ and 5′UTR of SIRP alpha transcripts. Taken together, we predict that miR-520d-5p and miR-520d-3p may be having role in the regulation of under-expressed SIRP alpha protein expression.

Keywords

Astrocytoma Glioblastoma SIRP alpha miRNA miR-20d-5p miR-520d-3p 
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Notes

Acknowledgements

The authors acknowledge KFRC, KIMS for the ethical permission, Dr. M. Sailaja, Head, Department of pathology, KIMS for diagnosis and histopathology of tissues and National Brain Repository, Brain Bank of NIMHANS, Bengaluru, India for the control samples.

Compliance with Ethical Standards

Funding

RDP thankful to Department of Biotechnology (DBT-India) (Award no: DBT JRF/2011-12/95) for student fellowship. Department of Science and technology (DST-India) (Grant no: SB/EMEQ-257/2013) and University with potential for excellence (UPE-India) (Grant no: UH/UGC/UPE-2 Interface studies/Research Projects/B1.4) for lab funding is acknowledged.

Conflict of Interest

The authors declare that they have no conflicts of interest.

Ethical Standard

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Ravindra Pramod Deshpande
    • 1
  • Y.B.V.K Chandra Sekhar
    • 2
  • Manas Panigrahi
    • 2
  • Phanithi Prakash Babu
    • 1
  1. 1.Department of Biotechnology and Bioinformatics, School of Life SciencesUniversity of HyderabadHyderabadIndia
  2. 2.Krishna Institute of Medical SciencesSecunderabadIndia

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