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Updated: Mar 3, 2026

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
Published on: July 21, 2017
[Effect of Autophagy on Homoharringtonine-treated K562 Cells]
Xue-Ying Lu1, Li-Lin Ye2, Yu-Ye Shi1
1Department of Hematology, Huaian First People's Hospital,Huaian 223300, Jiangsu Province, China.
Objective:
To study the effect of homoharringtonine (HHT) alone or combined with 3-methyladenine (3-MA) , an autophagy inhibitor, on the apoptosis and autophagy of K562 cells.
Methods:
K562 cells were treated with HHT(10 ng/ml) or HHT(10 ng/ml) combined with 3-MA (1.5 mmol/L) for 1 to 8 days. The apoptosis of treated cells was tested by means of flow cytometry(FCM), and the autophagy levels were tested with RT-PCR, Western blot and electron microscopy.
Results:
In the early stage of HHT-treated group, the apoptosis rate increased and decreased later. Beclin1 mRNA expression level and the LC3II/I ratio were declined firstly and increased later in HHT group. While combining with autophagy inhibitor 3-MA, both the Beclin1 mRNA expression level and the LC3II/I ratio were declined continually during the treated period. The activated caspase-3 protein expression level was also raised sustainability during both HHT and 3-MA cultured period.
Conclusions:
HHT can induce apoptosis of K562 cells, but the sustaining effect of HHT can induc autophagy of K562 cells, the combination of HHT with 3-MA may enhance the cytotoxicitic effect of HHT on K562 cells.
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