Identification of genes responsive to low intensity pulsed ultrasound in a human leukemia cell line Molt-4
Yoshiaki Tabuchi1, Hidetaka Ando, Ichiro Takasaki
1Division of Molecular Genetics, Life Science Research Center, University of Toyama, 2630 Sugitani, Toyama, Japan. ytabu@cts.u-toyama.ac.jp
Abstract:
We examined the gene expression of human leukemia Molt-4 cells treated with non-thermal low intensity pulsed ultrasound. Six hours after 0.3W/cm(2) pulsed ultrasound treatment, apoptosis (24+/-3.3%, mean+/-SD) with minimal cell lysis was observed. Of approximately 16,600 genes analyzed, BCL2-associated athanogene 3 (BAG3), DnaJ (Hsp40) homolog, subfamily B, member 1 (DNAJB1), heat shock 70 kDa protein 1B (HSPA1B), and heat shock 70 kDa protein 6 (HSPA6) showed increased levels of expression while isopentenyl-diphosphate delta isomerase (IDI1) and 3-hydroxy-3-methylglutaryl-coenzyme A synthase 1 (HMGCS1) showed decreased levels in the cells 3h after the ultrasound treatment. The expression levels of these six genes were confirmed by a real-time quantitative polymerase chain reaction. To our knowledge, this is the first report of DNA microarray analysis of genes that are differentially expressed in response to apoptosis induced by non-thermal low intensity pulsed ultrasound in human leukemia cells. The present results will provide a basis for further understanding of the molecular mechanisms of effects of not only low intensity pulsed ultrasound but also that of mechanical shear stress in the cells.
Insights
Non-thermal low intensity pulsed ultrasound induces apoptosis in human leukemia cells. Gene expression analysis revealed significant changes in apoptosis-related genes, offering insights into ultrasound
Area of Science:
- Biophysics
- Molecular Biology
- Cell Biology
Background:
- Non-thermal low intensity pulsed ultrasound (LIPUS) is a therapeutic modality with potential applications in cancer treatment.
- Understanding the molecular mechanisms underlying LIPUS-induced cell death is crucial for its clinical development.
- Human leukemia Molt-4 cells provide a relevant model for studying LIPUS effects on cancer cells.
Purpose of the Study:
- To investigate the gene expression profile of human leukemia Molt-4 cells following non-thermal LIPUS treatment.
- To identify specific genes involved in apoptosis induction by LIPUS.
- To elucidate the molecular pathways affected by LIPUS and mechanical shear stress in leukemia cells.
Main Methods:
- Human leukemia Molt-4 cells were exposed to non-thermal LIPUS at 0.3 W/cm(2).
- DNA microarray analysis was performed to assess genome-wide gene expression changes.
- Real-time quantitative polymerase chain reaction (qPCR) was used to validate the expression levels of key genes.
Main Results:
- Apoptosis was observed in Molt-4 cells six hours post-LIPUS treatment (24+/-3.3%) with minimal cell lysis.
- Microarray analysis of approximately 16,600 genes revealed differential expression patterns.
- Upregulated genes included BAG3, DNAJB1, HSPA1B, and HSPA6; downregulated genes included IDI1 and HMGCS1.
- qPCR confirmed the altered expression of these six genes three hours after LIPUS exposure.
Conclusions:
- This study presents the first DNA microarray analysis of gene expression changes in human leukemia cells induced by non-thermal LIPUS.
- LIPUS treatment significantly alters the expression of genes associated with apoptosis and cellular stress response.
- The findings provide a foundation for understanding the molecular effects of LIPUS and mechanical shear stress in leukemia cells.
