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Detection of Targetable Alterations in Non-small Cell Lung Cancer using Next-generation Sequencing
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Investigating Metabolically Altered Pathways in Small Cell Lung Cancer: From RNA Sequencing Analysis to

Subhadeep Das1, Sagar M Utturkar2, Roshnee Bose3

  • 1Department of Biotechnology, School of Life Science & Biotechnology, Adamas University, Kolkata 700126, India.

Methods and Protocols
|March 24, 2026
PubMed
Summary

Small cell lung cancer (SCLC) therapy targeting DDX5 with Supinoxin disrupts cellular respiration. This mitochondrial dysfunction offers new insights into SCLC treatment strategies.

Keywords:
DEAD-box helicase 5 (DDX5)RNA sequencingSeahorse XF Cell Mito Stress Testmitochondrial dysfunctionsmall cell lung cancer

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Area of Science:

  • Oncology
  • Molecular Biology
  • Biochemistry

Background:

  • Small cell lung cancer (SCLC) is aggressive, with high relapse rates due to chemoresistance.
  • The DEAD-box RNA helicase DDX5 is overexpressed in SCLC and is a potential therapeutic target.
  • Supinoxin (RX-5902) is a DDX5 inhibitor with demonstrated anti-tumor activity.

Purpose of the Study:

  • To investigate the molecular mechanisms underlying Supinoxin's anti-cancer effects in SCLC.
  • To explore the role of DDX5 in cellular respiration and energy metabolism in chemoresistant SCLC cells.
  • To validate the disruption of mitochondrial function as a key cytotoxic pathway for Supinoxin.

Main Methods:

  • Transcriptomic profiling using RNA sequencing (RNA-seq) to analyze gene expression changes.
  • Quantitative assessment of mitochondrial respiration using the Seahorse XF Cell Mito Stress Test.
  • Analysis of oxygen consumption rate (OCR) to evaluate mitochondrial bioenergetics.

Main Results:

  • RNA-seq revealed significant downregulation of genes critical for cellular respiration after Supinoxin treatment and DDX5 knockdown.
  • Seahorse analysis demonstrated marked mitochondrial dysfunction in Supinoxin-treated chemoresistant H69AR cells.
  • These results confirm that DDX5 inhibition by Supinoxin impairs cellular energy metabolism.

Conclusions:

  • DDX5 plays a significant role in regulating mitochondrial function in SCLC.
  • Supinoxin's cytotoxic effects are mediated, in part, by disrupting mitochondrial respiration.
  • These findings highlight DDX5 inhibition as a promising targeted therapy strategy for SCLC.