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IDENTIFICATION AND FUNCTIONAL CHARACTERIZATION OF MICRORNAS REGULATING HTERT.

Maham Ansari1, Rafiullah Rafiullah1, Abdul Wali1

  • 1Department of Biotechnology, Faculty of Life Sciences and Informatics, Balochistan University of Information Technology, Engineering and Management Sciences (BUITEMS), Quetta, Pakistan.

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This study identified specific microRNAs (miRNAs) that target human telomerase reverse transcriptase (hTERT). Inhibiting hTERT with miRNAs like hsa-miR-4651 reduced cancer cell proliferation, offering potential for targeted cancer therapies.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biochemistry

Background:

  • Telomerase, a ribonucleoprotein (RNP) reverse transcriptase, maintains genome integrity by replicating chromosome ends.
  • Inhibition of telomerase (hTERT) is a potential strategy for cancer prevention and treatment.
  • MicroRNAs (miRNAs) are explored as tools to target hTERT expression.

Purpose of the Study:

  • To investigate miRNA interactions with hTERT.
  • To determine the binding strength, affinity, and orientation of miRNAs targeting hTERT.
  • To verify the effect of miRNA on hTERT expression in cancer cells in vitro.

Main Methods:

  • Utilized miRWalk, TargetScan, and miRDB databases for miRNA screening.
  • Identified five top-hit miRNAs (hsa-miR-4651, hsa-miR-608, hsa-miR-6796-5p, hsa-miR-6752-5p, hsa-miR-6791-5p) targeting hTERT mRNA.
  • Employed in silico tools for miRNA-hTERT mRNA structure modeling, docking, and molecular dynamics (MD) simulations.

Main Results:

  • Selected miRNA expression was inhibited in MCF-7 breast cancer cells.
  • hsa-miR-4651 significantly reduced hTERT protein expression.
  • Inhibition of hsa-miR-4651 decreased melanoma and breast cancer cell proliferation.

Conclusions:

  • Established a procedure for identifying and verifying miRNA-mRNA interactions.
  • Demonstrated differential regulation of hTERT by specific miRNAs.
  • Showcased miRNA inhibition's potential to modulate hTERT expression and cell proliferation for targeted cancer therapy.
  • The strategy can be applied to screen miRNAs for other genes.