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KMID : 0366220150500020080
Korean Journal of Hematology
2015 Volume.50 No. 2 p.80 ~ p.86
Down-regulation of the autophagy gene, ATG7, protects bone marrow-derived mesenchymal stem cells from stressful conditions
Molaei Sedigheh

Roudkenar Mehryar Habibi
Amiri Fatemeh
Harati Mozhgan Dehghan
Bahadori Marzie
Jaleh Fatemeh
Jalili Mohammad Ali
Roushandeh Amaneh Mohammadi
Abstract
Background: Mesenchymal stem cells (MSCs) are valuable for cell-based therapy. However, their application
is limited owing to their low survival rate when exposed to stressful conditions. Autophagy, the process by which cells recycle the cytoplasm and dispose of defective organelles, is activated by stress stimuli to adapt, tolerate adverse conditions, or trigger the apoptotic machinery. This study aimed to determine whether regulation of autophagy
would affect the survival of MSCs under stress conditions.

Methods : Autophagy was induced in bone marrow-derived MSCs (BM-MSCs) by rapamycin, and was inhibited via shRNA-mediated knockdown of the autophagy specific gene, ATG7. ATG7 expression in BM-MSCs was evaluated by reverse transcription polymerase chain reaction (RT-PCR), western blot, and quantitative PCR (qPCR). Cells were then exposed to harsh microenvironments, and a water-soluble tetrazolium salt (WST)-1 assay was performed
to determine the cytotoxic effects of the stressful conditions on cells.

Results: Of 4 specific ATG7-inhibitor clones analyzed, only shRNA clone 3 decreased ATG7 expression. Under normal conditions, the induction of autophagy slightly increased the viability of MSCs while autophagy inhibition decreased their viability. However, under stressful conditions such as hypoxia, serum deprivation, and oxidative stress, the induction
of autophagy resulted in cell death, while its inhibition potentiated MSCs to withstand the stress conditions. The viability of autophagy-suppressed MSCs was significantly higher than that of relevant controls (P£¼0.05, P£¼0.01 and P£¼0.001).

Conclusion: Autophagy modulation in MSCs can be proposed as a new strategy to improve their survival rate in stressful microenvironments.
KEYWORD
Autophagy, Mesenchymal stem cells, Oxidative stress, Cell survival, shRNA
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