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Synergistic effect of metformin and vemurufenib (PLX4032) as a molecular targeted therapy in anaplastic thyroid cancer: an in vitro study

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Abstract

Background

Survival rate of patients affected with anaplastic thyroid carcinoma (ATC) is less than 5% with current treatment. In ATC, BRAFV600E mutation is the major mutation that results in the transformation of normal cells in to an undifferentiated cancer cells via aberrant molecular signaling mechanisms. Although vemurufenib is a selective oral drug for the BRAFV600E mutant kinase with a response rate of nearly 50% in metastatic melanoma, our study has showed resistance to this drug in ATC. Hence the rationale of the study is to explore combinational therapeutic effect to improve the efficacy of vemurafenib along with metformin. Metformin, a diabetic drug is an AMPK activator and has recently proved to be involved in preventing or treating several types of cancer.

Methods and results

Using iGEMDock software, a protein–ligand interaction was successful between Metformin and TSHR (receptor present in the thyroid follicular cells). Our study demonstrates that combination of vemurufenib with metformin has synergistic anti-cancer effects which was evaluated through MTT assay (cytotoxicity), colony formation assay (antiproliferation evaluation) and suppressed the progression of ATC cells growth by inducing significant apoptosis, proven by Annexin V-FITC assay (Early Apoptosis Detection). Downregulation of ERK signaling, upregulation of AMPK pathway and precision in epithelial-mesenchymal transition (EMT) pathway which were assessed by RT-PCR and Western blot provide the evidence that the combination of drugs involved in the precision of altered molecular signaling Further our results suggest that Metformin act as a demethylating agent in anaplastic thyroid cancer cells by inducing the expression of NIS and TSHR. Our study for the first time explored cAMP signaling in ATC wherein cAMP signaling is downregulated due to decrease in intracellular cAMP level upon metformin treatment.

Conclusion

To conclude, our findings demonstrate novel therapeutic targets and treatment strategies for undifferentiated ATC.

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Data availability

All data generated or analysed during this study are included in this published article.

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Acknowledgements

The authors deeply thank BRNS, DST-Women scientist, Government of India for providing financial support for this research work. We sincerely thank Dr. D. Karunagaran, Head, Department of Biotechnology, Bhupat & Jyoti Mehta School of Biosciences, IIT Madras for allowing the access to the laboratory.

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Authors and Affiliations

  1. Department of Preventive Oncology (Research), Cancer Institute, Adyar, Chennai, India

    Latha Durai, Soundharya Ravindran & Ramshankar Vijayalakshmi

  2. Department of Surgical Oncology, Cancer Institute, Adyar, Chennai, India

    Krishnamurthy Arvind

  3. Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, India

    Devarajan Karunagaran

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  1. Latha Durai
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  5. Ramshankar Vijayalakshmi
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Contributions

VR and LD designed the study. LD conducted the literature search of the study and performed the experiments. SR performed Bioinformatics work. The study was supervised by VR, DK, KA and KR. VR and LD analysed and validated the data and wrote the manuscript. All authors had read the manuscript and approved the publication of this manuscript.

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Correspondence to Ramshankar Vijayalakshmi.

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Durai, L., Ravindran, S., Arvind, K. et al. Synergistic effect of metformin and vemurufenib (PLX4032) as a molecular targeted therapy in anaplastic thyroid cancer: an in vitro study. Mol Biol Rep 48, 7443–7456 (2021). https://doi.org/10.1007/s11033-021-06762-7

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