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Targeted disruption of PI3K/Akt/mTOR signaling pathway, via PI3K inhibitors, promotes growth inhibitory effects in oral cancer cells

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Abstract

Purpose

The phosphoinositide-3-kinase (PI3K) pathway is the frequently altered in human cancer. This has led to the development and study of novel PI3K inhibitors for targeted therapy and also to overcome resistance to radiotherapy.

Method

The anti-tumour effects of PI3K inhibitors (PI-828, PI-103 and PX-866) in terms of cell proliferation, colony formation, induction of apoptosis, cell cycle arrest, invasion, autophagy, and pNF-κB/p65 translocation in SCC-4, SCC-9 and SCC-25 cells were studied by performing MTT, clonogenic, DAPI staining, propidium iodide staining, annexin-V binding, matrigel invasion, acridine orange staining and immuno-fluorescence assay. Western blot assay was performed to assess the alteration in the expression of various proteins.

Result

PI-828 and PI-103 treatment exhibited dose-dependent inhibition of growth and proliferation of OSCC cells with a concomitant induction of apoptosis, altered cell cycle regulation and decreased invasiveness (p < 0.01). PX-866 induced apoptosis, cell cycle arrest, autophagy and a significant decrease in the invasiveness of oral cancer cells as compared to untreated cells (p < 0.01). These compounds significantly reduced expression of COX-2, cyclin-D1 and VEGF in the treated cells besides cytoplasmic accumulation of pNF-κB/p65 protein. In addition to PI3Kα, inactivation of downstream components, i.e. Akt and mTOR was seen.

Conclusion

PI3K inhibitors such as PI-103, PI-828 and PX-866 may be developed as potential therapeutic agents for effective treatment of oral squamous cell carcinoma (OSCC) patients, associated with activated PI3K/Akt pathway.

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Acknowledgements

The study was funded by Department of Science and Technology (DST), Government of India, under SERB Program. SA was recipient of senior research fellowship from Department of Science and Technology (DST), and SJ and LS were recipient of studentship from Department of Biotechnology, Government of India.

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Author notes

  1. Sadhna Aggarwal and Sarah John: equal contribution as first author.

Authors and Affiliations

  1. Department of Biotechnology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India

    Sadhna Aggarwal, Sarah John, Leena Sapra & Satya N. Das

  2. Department of Otorhinolaryngology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India

    Suresh C. Sharma

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  1. Sadhna Aggarwal
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  2. Sarah John
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Correspondence to Satya N. Das.

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Electronic supplementary material

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280_2018_3746_MOESM1_ESM.ppt

Supplementary Fig. 1: (a) Photomicrographs and (b) bar graphs as obtained after In vitro clonogenic assay indicating growth inhibitory effects of PI3K inhibitors (IC 50) treatment on oral cancer cell lines (SCC-4, SCC-9 and SCC-25 cells) (PPT 1397 KB)

280_2018_3746_MOESM2_ESM.ppt

Supplementary Fig. 2: Representative photomicrographs showing nuclear DNA fragmentation of PI-103, PI-828 and PX-866 treated SCC-4 cells in comparison to normal nuclear DNA in untreated SCC-4 cells (Blue: DAPI stain) (PPT 1629 KB)

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Aggarwal, S., John, S., Sapra, L. et al. Targeted disruption of PI3K/Akt/mTOR signaling pathway, via PI3K inhibitors, promotes growth inhibitory effects in oral cancer cells. Cancer Chemother Pharmacol 83, 451–461 (2019). https://doi.org/10.1007/s00280-018-3746-x

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  • DOI: https://doi.org/10.1007/s00280-018-3746-x

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