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Identification and Characterization of Cancer Stem-Like Cells in ALK-Positive Anaplastic Large Cell Lymphoma Using

Jing Li1,2, Moinul Haque1,3, Chuquan Shang1

  • 1Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB T6G 2E1, Canada.

Current Issues in Molecular Biology
|July 21, 2021
PubMed
Summary

The SORE6 reporter system effectively identifies and enriches cancer stem-like (CSL) cells in ALK-positive anaplastic large cell lymphoma (ALK+ ALCL). These identified CSL cells exhibit increased chemoresistance and clonogenicity, driven by Sox2 and Oct4 transcription factors.

Keywords:
Oct4SORE6Sox2anaplastic large cell lymphomacancer stem-like cells

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Area of Science:

  • Oncology
  • Stem Cell Biology
  • Molecular Biology

Background:

  • Transcription factors Sox2 and Oct4 are crucial for maintaining pluripotency in embryonic stem cells and conferring stemness in cancer stem-like (CSL) cells.
  • Identifying and isolating CSL cells is vital for understanding cancer progression and developing targeted therapies, particularly in non-hematologic malignancies.

Purpose of the Study:

  • To develop and validate the SORE6 in-vitro reporter system for quantifying Sox2/Oct4 transcription activity.
  • To identify and enrich CSL cells within ALK-positive anaplastic large cell lymphoma (ALK+ ALCL) using the SORE6 system.

Main Methods:

  • Utilized the SORE6 reporter system to identify and isolate SORE6+ cells from ALK+ ALCL cell lines (SupM2, UCONN-L2).
  • Performed functional assays including chemoresistance and colony-formation assays on single-cell clones derived from SORE6+ and SORE6- cells.
  • Conducted shRNA knockdown of Sox2 and enforced expression of Sox2/Oct4 to assess their role in CSL properties.
  • Analyzed protein expression and subcellular localization via Western blots and assessed Sox2/Oct4 binding to the SORE6 consensus sequence.

Main Results:

  • The SORE6 system successfully identified and enriched CSL cells in ALK+ ALCL, with approximately 20% of cells being SORE6+.
  • SORE6+ cells demonstrated significantly higher chemoresistance and clonogenic potential compared to SORE6- cells.
  • Knockdown of Sox2 reduced chemoresistance in SORE6+ cells, while enforced Sox2/Oct4 expression increased these properties in SORE6- cells.
  • Sox2 and Oct4 directly bind to the SORE6 consensus sequence in SORE6+ cells, and c-Myc regulates SORE6 activity.

Conclusions:

  • The SORE6 reporter system is an effective tool for identifying and enriching cancer stem-like cells in ALK-positive anaplastic large cell lymphoma.
  • Sox2 and Oct4 play a direct role in conferring chemoresistance and clonogenicity, key characteristics of CSL cells in ALK+ ALCL.
  • The SORE6 system facilitates further investigation into the mechanisms driving cancer stemness in ALK+ ALCL.