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Viability Assays for Cells in Culture
Published on: January 20, 2014
[Effect of proteasome inhibitors MG-132 at different doses on cultured K562 cell apoptosis]
Qing-Wen Shao1, Tian-Yang Xue, Ji-Zhao Gao
1Department of Pediatrics, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangshu Province, China.
Abstract:
This study was aimed to investigate the effect of proteasome inhibitor MG-132 at different doses on cultured K562 cell apoptosis. MTT assay was used to observe the activity of K562 cell proliferation inhibition rate after treating for 48 hours at different doses (0, 2, 4, 8, 16, 32 micromol/L). Immunocytochemistry was used to detect the NF-kappaB activity and glucocorticoid receptor (GR) expression. Flow cytometry was used to determine the K562 cell apoptosis. The results indicated that proliferation inhibition rate of K562 cells after treated for 48 hours showed dose-dependent, the inhibitory rates of cell proliferation in test groups were significant higher than that in control group, and the effect in 32 micromol/L test group was the most obvious (45.24 +/- 4.12)% (p < 0.05). The NF-kappaB activity and GR expression after treating for 48 hours showed dose-dependent. Compared with control group, the NF-kappaB activities in test groups were lower (p < 0.05), and the NF-kappaB activity in 32 micromol/L test group was the lowest (63.60 +/- 2.95); the GR expression in test groups was higher (p < 0.05), and the GR expression in 16 micromol/L test group was the highest (75.62 +/- 2.70). The K562 cell apoptosis rate after treating for 48 hours also showed dose-dependent. Compared with control group, the K562 cell apoptosis rates in test groups were higher (p < 0.05), the K562 cell apoptosis rate in 32 micromol/L test group was the highest (21.37 +/- 2.02)%. It is concluded that the MG-132 may induce K562 cell apoptosis and proliferation inhibition through up-regulation of NF-kappaB activity and down-regulation of GR expression both in dose-dependent manner.
Insights
Proteasome inhibitor MG-132 induces K562 cell apoptosis and proliferation inhibition in a dose-dependent manner. This occurs through decreased NF-kappaB activity and increased glucocorticoid receptor (GR) expression.
Area of Science:
- Cell Biology
- Pharmacology
- Cancer Research
Background:
- K562 cells are a human chronic myeloid leukemia cell line.
- Proteasome inhibitors are a class of drugs that block proteasome activity.
- NF-kappaB and glucocorticoid receptor (GR) play roles in cell survival and apoptosis.
Purpose of the Study:
- To investigate the effects of proteasome inhibitor MG-132 on K562 cell apoptosis.
- To determine the dose-dependent relationship of MG-132's effects.
- To examine the impact of MG-132 on NF-kappaB activity and GR expression.
Main Methods:
- MTT assay for cell proliferation inhibition.
- Immunocytochemistry for NF-kappaB activity and GR expression.
- Flow cytometry for K562 cell apoptosis analysis.
Main Results:
- MG-132 demonstrated a dose-dependent inhibition of K562 cell proliferation.
- NF-kappaB activity decreased, while GR expression increased with higher MG-132 doses.
- K562 cell apoptosis rates significantly increased in a dose-dependent manner with MG-132 treatment.
Conclusions:
- MG-132 induces K562 cell apoptosis and proliferation inhibition.
- These effects are mediated by a dose-dependent down-regulation of NF-kappaB activity and up-regulation of GR expression.
- MG-132 shows potential as a therapeutic agent for K562 leukemia.

