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[Effects of down-regulated TRAF6 gene expression on the proliferation and apoptosis in multiple myeloma cells]
Hong-ming Huang1, Xing-feng Wang, Xin-xin Liu
1Affiliated Hospital of Nantong University, Nantong 226001, China.
Objective:
To investigate the down-regulated TRAF6 gene expression and its effects on proliferation and apoptosis in multiple myeloma (MM) cells.
Methods:
Detection of TRAF6 expression were conducted by RT-PCR and Western blot in MM cell lines of KM3, U266, RPMI8226 and primary cells from patients. RPMI8226 cell lines were transfected with siRNA of TRAF6. The efficiency of transfection was identified by using of fluorescence microscope, RT-PCR, and Western blot. The levels of proliferation were analyzed by CCK-8 method under the different concentrations of siRNA. Apoptosis rate were detected with Hoechst33258/PI double staining by flow cytometry. Apoptosis related proteins Bcl-2, BAX, and NF-κB signal pathway were observed before and after siRNA transfection by Western blot.
Results:
The levels of TRAF6 mRNA and protein in MM cell lines, especially in primary myeloma cells, were significantly higher than those in controls. After transfected with 50 nmol/L siRNA in RPMI8226 cells, the relative level of TRAF6 mRNA (0.49±0.24) was significantly lower than that in non-transfected group (1.87±0.23) and idling group (1.74±0.35). The proliferation rate of siRNA transfected cells decreased with dose dependence (P<0.01). The apoptosis rates increased from 11.20% (before transfection) to 51.82% (after transfection), accompanied by down-regulated Bcl-2 protein, NF-κB signal pathway (p-p65 and p52), and up-regulated BAX protein.
Conclusion:
TRAF6 expression was high in myeloma cells. TRAF6 siRNA could inhibit proliferation of myeloma cells and induce apoptosis mediated by NF-κB classical and alternative pathway in myeloma cells.
Insights
High TRAF6 gene expression fuels multiple myeloma (MM) cell growth. Inhibiting TRAF6 with siRNA significantly reduces MM cell proliferation and triggers apoptosis via the NF-κB pathway.
Area of Science:
- Molecular Biology
- Cancer Research
- Immunology
Context:
- Multiple myeloma (MM) is a hematological malignancy characterized by uncontrolled proliferation of plasma cells.
- TRAF6 (TNF receptor-associated factor 6) is implicated in various cellular processes, including immune response and cell survival.
- Dysregulation of TRAF6 expression has been observed in several cancers, suggesting its potential role in tumorigenesis.
Purpose:
- To investigate the role of Tumor Necrosis Factor Receptor Associated Factor 6 (TRAF6) gene expression in multiple myeloma (MM) cells.
- To determine the effects of TRAF6 down-regulation on MM cell proliferation and apoptosis.
- To elucidate the underlying molecular mechanisms involving the NF-κB signaling pathway.
Summary:
- TRAF6 mRNA and protein levels were significantly elevated in MM cell lines and primary patient cells compared to controls.
- Transfection of RPMI8226 MM cells with TRAF6 siRNA dose-dependently inhibited cell proliferation.
- TRAF6 siRNA treatment induced significant apoptosis, decreasing Bcl-2, up-regulating BAX, and modulating the NF-κB signaling pathway (p-p65, p52).
Impact:
- This study identifies TRAF6 as a potential therapeutic target in multiple myeloma.
- Down-regulation of TRAF6 presents a promising strategy for inhibiting MM cell proliferation and inducing apoptosis.
- Understanding TRAF6's role in MM pathogenesis provides insights into novel treatment approaches targeting the NF-κB pathway.
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