The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
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ATG5 knockout promotes paclitaxel sensitivity in drug-resistant cells via induction of necrotic cell death

  • Hwang, Sung-Hee (Division of Life Sciences, College of Life Sciences and Bioengineering, Incheon National University) ;
  • Yeom, Hojin (Division of Life Sciences, College of Life Sciences and Bioengineering, Incheon National University) ;
  • Lee, Michael (Division of Life Sciences, College of Life Sciences and Bioengineering, Incheon National University)
  • Received : 2019.11.01
  • Accepted : 2020.02.10
  • Published : 2020.05.01
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Abstract

Autophagy regulators are often effective as potential cancer therapeutic agents. Here, we investigated paclitaxel sensitivity in cells with knockout (KO) of ATG5 gene. The ATG5 KO in multidrug resistant v-Ha-ras-transformed NIH 3T3 cells (Ras-NIH 3T3/Mdr) was generated using the CRISPR/Cas9 technology. The qPCR and LC3 immunoblot confirmed knockout of the gene and protein of ATG5, respectively. The ATG5 KO restored the sensitivity of Ras-NIH 3T3/Mdr cells to paclitaxel. Interestingly, ATG5 overexpression restored autophagy function in ATG5 KO cells, but failed to rescue paclitaxel resistance. These results raise the possibility that low level of resistance to paclitaxel in ATG5 KO cells may be related to other roles of ATG5 independent of its function in autophagy. The ATG5 KO significantly induced a G2/M arrest in cell cycle progression. Additionally, ATG5 KO caused necrosis of a high proportion of cells after paclitaxel treatment. These data suggest that the difference in sensitivity to paclitaxel between ATG5 KO and their parental MDR cells may result from the disparity in the proportions of necrotic cells in both populations. Thus, our results demonstrate that the ATG5 KO in paclitaxel resistant cells leads to a marked G2/M arrest and sensitizes cells to paclitaxel-induced necrosis.

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