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Galectin-3-induced cell spreading and motility relies on distinct signaling mechanisms compared to fibronectin

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Abstract

Secreted galectin-3 often gets incorporated into extracellular matrix and is utilized by cancer cells for spreading, movement, and metastatic dissemination. Here we investigate molecular mechanisms by which galectin-3 brings about these effects and compare it with fibronectin. Imaging of cells spread on fibronectin showed stress fibers throughout cell body, however, galectin-3-induced formation of parallel actin bundles in the lamellipodial region resulting in unique morphological features. FRAP analysis showed that the actin turnover in the lamellipodial region was much higher in cells spread on galectin-3 as compared to that on fibronectin. Rac1 activation is correlated with lamellipodial organization on both the substrates. Activation of Akt and Rac1, the regulators of actin dynamics, show inverse correlation with each other on both galectin-3 and fibronectin. Activation of Erk however, remained similar. Further, inhibition of activation of Akt and Erk inhibited spreading and motility of cells on galectin-3 but not on fibronectin. The results very comprehensively demonstrate distinct signaling pathways that regulate microfilament organization, lamellipodial structures, spreading, and movement of cells plated on galectin-3 as opposed to fibronectin.

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Acknowledgments

We dedicate this work to the memory of Late Dr. Rajiv D. Kalraiya, who was responsible for planning and supervision of this work. We acknowledge Dr. Hakon Leffler for providing E. coli BL21 star strain expressing recombinant human galectin-3 and Dr. Sorab Dalal for providing E. coli strain expressing GST-PAK1 fusion protein. We acknowledge Urjita Joshi for signaling pathway experiments, Vaishali Khailaje, Tanuja Dighe, and Jairaj Kashale for help in microscopy experiments, and D. S. Chavan and A. M. Pawar for the technical help during galectin-3 purification. We thank Council of Scientific and Industrial Research (CSIR), India for providing the fellowship to Shyam K. More and Department of Biotechnology (DBT), India for funding the project.

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Correspondence to Shubhada V. Chiplunkar.

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The authors declare that they have no conflict of interest.

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Rajiv D. Kalraiya: Deceased.

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More, S.K., Chiplunkar, S.V. & Kalraiya, R.D. Galectin-3-induced cell spreading and motility relies on distinct signaling mechanisms compared to fibronectin. Mol Cell Biochem 416, 179–191 (2016). https://doi.org/10.1007/s11010-016-2706-1

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Keywords

  • Galectin-3
  • Fibronectin
  • Microfilaments
  • Lamellipodia
  • Cell spreading
  • Cell migration