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Updated: May 7, 2026
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Published on: February 28, 2017
NOTCH knockdown affects the proliferation and mTOR signaling of leukemia cells
Yuki Okuhashi1, Mai Itoh, Nobuo Nara
1Department of Laboratory Medicine, Tokyo Medical and Dental University, Yushima 1-5-45, Bunkyo-Ku, Tokyo 113-8519, Japan. tohda.mlab@tmd.ac.jp.
Aim:
The effects of small interfering RNA (siRNA)-mediated knockdown of NOTCH1 and NOTCH2 on cell proliferation and downstream signaling pathways in leukemia cells were examined.
Materials And Methods:
Two T-lymphoblastic leukemia (T-ALL) cell lines and two acute myeloblastic leukemia (AML) cell lines were transfected with siRNAs targeting NOTCH1 and NOTCH2. The effects of knockdown on cell proliferation and protein expression were examined by colorimetric WST-8 assay and immunoblotting, respectively.
Results:
In T-ALL cell lines, NOTCH1 knockdown as well as NOTCH2 knockdown suppressed cell proliferation and induced apoptosis. v-Myc avian myelocytomatosis viral oncogene homolog (MYC) protein expression was down-regulated in NOTCH1-knockdown cells but not affected in NOTCH2-knockdown cells. In AML cell lines, cell proliferation was not significantly affected by NOTCH siRNAs. NOTCH2 knockdown increased the level of cleaved NOTCH1 fragment without increasing NOTCH1 expression. NOTCH knockdown reduced the level of mechanistic target of rapamycin (mTOR) protein in the monoblastic leukemia cell line THP-1. Contrastingly, NOTCH activation by NOTCH ligand stimulation increased the expression of mTOR in THP-1 cells.
Conclusion:
These novel findings on NOTCH signaling may contribute to the development of effective NOTCH-targeted therapies against leukemia.
Insights
NOTCH1 and NOTCH2 knockdown suppressed leukemia cell proliferation and induced apoptosis in T-ALL cells, but not AML cells. NOTCH signaling impacts MYC and mTOR pathways, suggesting potential for targeted leukemia therapies.
Area of Science:
- Oncology
- Molecular Biology
- Cell Signaling
Background:
- NOTCH signaling is implicated in various cancers, including leukemia.
- Understanding NOTCH pathway roles in different leukemia subtypes is crucial for therapeutic development.
Purpose of the Study:
- To investigate the impact of NOTCH1 and NOTCH2 knockdown on leukemia cell proliferation and signaling.
- To explore the therapeutic potential of targeting NOTCH in T-lymphoblastic leukemia (T-ALL) and acute myeloblastic leukemia (AML).
Main Methods:
- Utilized small interfering RNA (siRNA) to knockdown NOTCH1 and NOTCH2 in T-ALL and AML cell lines.
- Assessed cell proliferation using WST-8 assay and protein expression via immunoblotting.
Main Results:
- NOTCH1 and NOTCH2 knockdown inhibited proliferation and induced apoptosis in T-ALL cells.
- NOTCH1 knockdown reduced MYC expression; NOTCH2 knockdown affected cleaved NOTCH1 levels.
- NOTCH knockdown reduced mTOR in THP-1 cells, while NOTCH activation increased mTOR.
Conclusions:
- NOTCH signaling plays a differential role in T-ALL versus AML.
- Targeting NOTCH1/NOTCH2 may offer a therapeutic strategy for T-ALL.
- Further research into NOTCH-mTOR interactions is warranted for leukemia treatment.
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