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Autophagy inhibition with chloroquine reverts paclitaxel resistance and attenuates metastatic potential in human nonsmall lung adenocarcinoma A549 cells via ROS mediated modulation of β-catenin pathway

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Abstract

Paclitaxel is one of the most commonly used drugs for the treatment of nonsmall cell lung cancer (NSCLC). However acquired resistance to paclitaxel, epithelial to mesenchymal transition and cancer stem cell formation are the major obstacles for successful chemotherapy with this drug. Some of the major reasons behind chemoresistance development include increased ability of the cancer cells to survive under stress conditions by autophagy, increased expression of drug efflux pumps, tubulin mutations etc. In this study we found that inhibition of autophagy with chloroquine prevented development of paclitaxel resistance in A549 cells with time and potentiated the effect of paclitaxel by increased accumulation of superoxide-producing damaged mitochondria, with elevated ROS generation, it also increased the apoptotic rate and sub G0/ G1 phase arrest with time in A549 cells treated with paclitaxel and attenuated the metastatic potential and cancer stem cell population of the paclitaxel-resistant cells by ROS mediated modulation of the Wnt/β-catenin signaling pathway, thereby increasing paclitaxel sensitivity. ROS here played a crucial role in modulating Akt activity when autophagy process was hindered by chloroquine, excessive ROS accumulation in the cell inhibited Akt activity. In addition, chloroquine pre-treatment followed by taxol (10 nM) treatment did not show significant toxicity towards non-carcinomas WI38 cells (lung fibroblast cells). Thus autophagy inhibition by CQ pre-treatment can be used as a fruitful strategy to combat the phenomenon of paclitaxel resistance development as well as metastasis in lung cancer.

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  • 28 March 2019

    The original version of this article unfortunately contained an error in acknowledgment text. The authors would like to include a statement: “Moumita Dasgupta is supported by Junior Research Fellowship from University Grant Commission, India.” in acknowledgment section.

Abbreviations

NSCLC:

Non small cell lung cancer

ROS:

Reactive oxygen species

Tx:

Paclitaxel/taxol

MDR1:

Multidrug resistant protein 1

MRP1:

Multidrug resistance-associated protein 1

Pgp:

P-glycoprotein

EMT:

Epithelial to mesenchymal transition

ROS:

Reactive oxygen species

CQ:

Chloroquine

DMEM:

Dulbecco’s modified eagle’s Media

DMSO:

Dimethyl sulfoxide

MDC:

Monodancyl cadaverine

MSR:

Mitosox red

OCR:

Oxygen consumption rate

ECAR:

Extracellular acidification rate

PE:

Phycoerythrin

CSC:

Cancer stem cell

NAC:

N-acetyl-cysteine

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Acknowledgements

The authors wish to thank Dr. Sib Sankar Roy, CSIR – Indian Institute of Chemical Biology, Kolkata, India for helping the use of Seahorse XFe24 Extracellular Flux Analyser facility. The work was supported by grants from Department of Science and Technology, Govt. of India (No. SR/SO/BB-14/2008) and Department of Biotechnology, Government of India (No. BT/ PR12889/AGR/36/624/2009) to G.Chakbarati. FACS and fluorescence microscope instruments facility were developed by grants from National Common Minimum Project, Government. of India. Confocal microscope instrument facility was developed by grants from DBT-IPLS facility. S Datta and D Das Mukherjee were supported by Senior Research fellowship from Council of Scientific and industrial research (CSIR), Government. of India. S. Bandopadhyay is supported by Junior research fellowship from Council of Scientific and industrial research (CSIR), India.

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Authors and Affiliations

  1. Department of Biotechnology and Dr. B. C. Guha Centre for Genetic Engineering and Biotechnology, University of Calcutta, 35 Ballygunge Circular Road, Ballygunge, Kolkata, West Bengal, 700 019, India

    Satabdi Datta, Dipanwita Das Mukherjee, Moumita Dasgupta & Gopal Chakrabarti

  2. School of Chemistry and Biochemistry, Thapar Institute of Engineering and Technology, Bhadson Road, Patiala, Punjab, 147004, India

    Diptiman Choudhury

  3. Department of Chemistry, National Institute of Technology, Ravangla, Sikkim, 737139, India

    Amlan Das

  4. Cell Biology & Physiology Division, CSIR – Indian Institute of Chemical Biology, Kolkata, 700032, India

    Shreya Bandopadhyay

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  1. Satabdi Datta
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Datta, S., Choudhury, D., Das, A. et al. Autophagy inhibition with chloroquine reverts paclitaxel resistance and attenuates metastatic potential in human nonsmall lung adenocarcinoma A549 cells via ROS mediated modulation of β-catenin pathway. Apoptosis 24, 414–433 (2019). https://doi.org/10.1007/s10495-019-01526-y

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