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[Effect of MELK Inhibitor OTSSP167 on Diffuse Large B-Cell Lymphoma]
Jun-Yi Zhou1, Hao Huang1, Yan Zhuang1
1The Huaian Clinical College of Xuzhou Medical University, Huai'an 223300, Jiangsu Province, China.
Objective:
To investigate the effect of MELK inhibitor OTSSP167 against diffuse large B-cell lymphoma (DLBCL).
Methods:
The effect of OTSSP167 on activity, proliferation, and apoptosis of DLBCL cell line (SUDHL2 and HBL1) was detected by CCK-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) staining, and Annexin V-FITC/PI double staining, respectively. DLBCL cells were inoculated into nude mice, after 4 weeks of OTSSP167 treatment, the effect of OTSSP167 on DLBCL growth in vivo was detected. Caspase-GloTM 3/7 enzyme activity assay kit was used to detect the effect of OTSSP167 on Caspase 3/7 enzyme activity of DLBCL cells. The expression levels of apoptosis and cycle-related proteins were detected by Western blot.
Results:
OTSSP167 significantly inhibited the activity of SUDHL2 and HBL1 cells in a dose-dependent manner (r =-0.61, r =-0.52). EdU staining showed that OTSSP167 could significantly inhibit the proliferation of SUDHL2 and HBL1 cells. Annexin V-FITC/PI result showed that OTSSP167 could significantly promote the apoptosis of SUDHL2 and HBL1 cells (P <0.001). The result of in vivo experiment showed that OTSSP167 could inhibit the growth of SUDHL2 cells in nude mice. The result of TUNEL staining of tumor further confirmed that OTSSP167 could promote the apoptosis of SUDHL2 cells. Caspase 3/7 enzyme activity test demonstrated that OTSSP167 could significantly increase caspase activity in SUDHL2 and HBL1 cells (r =0.98, r =0.87). Western blot showed that OTSSP167 could dose-dependently inhibit the expression of PARP, Bcl-xL, and Bcl-2 in apoptosis signaling pathway (r =-0.93, r =-0.66, r =-0.87), while p53 protein was significantly up-regulated (r =0.82). The expression of cell cycle-related proteins cdc2, Cyclin E1, Cyclin A2, and Cyclin B1 also showed a dose-dependent down-regulation (r =-0.89, r =-0.83, r =-0.61, r =-0.93).
Conclusion:
The MELK inhibitor OTSSP167 can inhibit the proliferation and promote the apoptosis of DLBCL cells by inhibiting the expression of cycle-related proteins and anti-apoptosis-related proteins.
Insights
The MELK inhibitor OTSSP167 effectively reduces diffuse large B-cell lymphoma (DLBCL) cell activity and proliferation while promoting apoptosis. This compound shows promise in inhibiting DLBCL growth both in vitro and in vivo.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Context:
- Diffuse large B-cell lymphoma (DLBCL) is an aggressive non-Hodgkin lymphoma with unmet therapeutic needs.
- Aberrant signaling pathways, including those involving MELK (maternalनल-enriched lineage kinase), are implicated in DLBCL pathogenesis.
- Targeting key regulatory proteins offers a potential strategy for novel DLBCL therapies.
Purpose:
- To evaluate the anti-cancer efficacy of the MELK inhibitor OTSSP167 against DLBCL cell lines (SUDHL2 and HBL1).
- To investigate the effects of OTSSP167 on DLBCL cell proliferation, apoptosis, and cell cycle progression.
- To assess the in vivo therapeutic potential of OTSSP167 in a DLBCL xenograft mouse model.
Summary:
- OTSSP167 significantly inhibited DLBCL cell activity and proliferation in a dose-dependent manner, as evidenced by CCK-8 and EdU assays.
- Treatment with OTSSP167 markedly promoted apoptosis in DLBCL cells, confirmed by Annexin V/PI staining, TUNEL assays, and increased Caspase 3/7 activity.
- Western blot analysis revealed that OTSSP167 down-regulated anti-apoptotic proteins (PARP, Bcl-xL, Bcl-2) and cell cycle regulators (cdc2, Cyclin E1, Cyclin A2, Cyclin B1), while up-regulating p53.
Impact:
- OTSSP167 demonstrates potent anti-proliferative and pro-apoptotic effects on DLBCL cells, suggesting its potential as a therapeutic agent.
- The study elucidates the molecular mechanisms by which OTSSP167 exerts its anti-cancer effects, involving modulation of key apoptosis and cell cycle proteins.
- These findings support further investigation of OTSSP167 as a targeted therapy for DLBCL and potentially other hematological malignancies.
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