Realgar transforming solution suppresses KG-1a-derived CD34+CD38- acute myeloid leukemia stem cell-like phenotypes in association with ER-mitochondrial stress and mitophagy-related alterations

  • J Ethnopharmacol. 2026 Oct 28:369:121896. doi: 10.1016/j.jep.2026.121896.
Teng Wang  1 ,  Weilin Liu  1 ,  Chunyi Lyu  2 ,  Yunxiao Luo  1 ,  Xinyu Tang  1 ,  Chen Han  1 ,  Ruirong Xu  3
Affiliations
  • 1. The First Clinical Medical College, Shandong University of Traditional Chinese Medicine, Jinan, Shandong, 250014, China.
  • 2. Key Laboratory of Integrated Traditional Chinese and Western Medicine for Hematology, Health Commission of Shandong Province, Jinan, Shandong, 250014, China; Institute of Hematology, Shandong University of Traditional Chinese Medicine, Jinan, Shandong, 250014, China.
  • 3. Key Laboratory of Integrated Traditional Chinese and Western Medicine for Hematology, Health Commission of Shandong Province, Jinan, Shandong, 250014, China; Institute of Hematology, Shandong University of Traditional Chinese Medicine, Jinan, Shandong, 250014, China; Department of Hematology, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong, 250014, China. Electronic address: [email protected].
PMID: 42184909 DOI: 10.1016/j.jep.2026.121896
Abstract

Ethnopharmacological relevance: Realgar is an arsenic-containing mineral medicine and a key component of Realgar-Indigo naturalis formula, an oral traditional Chinese medicine preparation clinically used in China for acute promyelocytic leukemia (APL), a biologically distinct subtype of acute myeloid leukemia (AML). Realgar Transforming Solution (RTS) is a microbially processed realgar preparation. Its activity in AML stem cell-like contexts outside APL remains insufficiently defined, particularly in cell-line-derived CD34+CD38- LSC-like models.

Aim of the study: To evaluate whether RTS suppresses stemness-associated phenotypic features in KG-1a-derived CD34+CD38- LSC-like cells and to explore its association with endoplasmic reticulum stress (ERS), mitochondrial injury, and mitophagy-related alterations.

Materials and methods: CD34+CD38- LSC-like cells were enriched from KG-1a cells and used as a cell-line-derived in vitro model. Cell viability, Apoptosis, cell cycle, CD34+CD38- fraction, colony formation, and Tim3/HAVCR2 expression were assessed after RTS treatment. RNA Sequencing, CA2+ measurement, ROS detection, mitochondrial membrane potential assay, MDC staining, western blotting, RT-qPCR, immunofluorescence, and transmission electron microscopy were performed. 4-phenylbutyric acid and Mdivi-1 were used as pharmacological rescue agents.

Results: RTS reduced the CD34+CD38- fraction, colony formation, and Tim3/HAVCR2 expression, while inducing Apoptosis in KG-1a-derived LSC-like cells. Transcriptomics implicated ER protein processing, unfolded protein response, Apoptosis, Autophagy, and mitophagy-related pathways. RTS increased CA2+ levels, ROS accumulation, mitochondrial depolarization, PERK-eIF2α-ATF4-CHOP signaling, and PINK1/Parkin-associated mitophagy-like marker changes. 4-phenylbutyric acid and Mdivi-1 partially rescued RTS-impaired LSC-like phenotypic features, supporting pathway involvement.

Conclusions: RTS suppressed stemness-associated phenotypic features in a KG-1a-derived CD34+CD38- LSC-like model. These effects were associated with ERS, mitochondrial injury, and PINK1/Parkin-related mitophagy-like alterations.

Keywords
Acute myeloid leukemia; Endoplasmic reticulum stress; Leukemia stem cell-like cells; Mitophagy-related alterations; Realgar transforming solution.
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