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The elevated activation of NFκB and AP-1 is correlated with differential regulation of Bcl-2 and associated with oral squamous cell carcinoma progression and resistance

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Abstract

Objectives

Oral cancer is the sixth most common cancer in the world. Failure of chemoradiation therapy is a major concern for treating oral cancer patients. The objective of this study is to determine the B cell lymphoma-2 (bcl-2) expression and its regulation by nuclear factor κB (NFκB) and activator protein 1 (AP-1) in oral cancer progression and chemoradiation resistance.

Materials and methods

In the present study, a total of 123 (n = 123) human samples were included. Briefly, 64 fresh samples were from adjacent normal (AN), primary oral tumors without treatment (PT), and tumors with resistance to chemoradiation therapy with local recurrence (RCRT). Fifty-nine samples were human tongue cancers and normal samples (TMA). Messenger RNA (mRNA) expression levels of bcl-2 and protein levels of bcl-2, NFκB, AP-1, and inactive GSK3α/β were measured by semiquantitative RT-PCR, immunohistochemistry, Western blot, and ChIP analysis.

Results

Increased bcl-2 expression was observed in PT compared to AN. The RCRT tumors showed maximum expression of bcl-2 mRNA and protein over the PT and AN groups. Bcl-2 protein and mRNA expression were positively correlated with NFκB and AP-1 expression. AP-1 expression was strongly correlated with bcl-2 in the RCRT group of tumors. Further, inactive GSK3α/β showed a positive trend with bcl-2 expression in oral tongue cancer specimens.

Conclusion

Collectively, our results demonstrated cumulative effect of AP-1 and NFĸB for bcl-2 gene regulation in overall PT progression and chemoradiation resistance. The study provides evidence of increased bcl-2 mRNA/protein fueled by NFĸB in PT and AP-1 in RCRT. These regulations of bcl-2 by NFκB and AP-1 are important in OSCC progression and chemoradiation resistance.

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Acknowledgements

The authors wish to acknowledge Prof. M.K. Rai (Pathologist), Director, RIMS, Ranchi and Prof. NK Jha, Head Dept. of Surgery (and his colleagues) RIMS, Ranchi; and the Director of CARA, Cancer Hospital, Ranchi and his colleagues Dr. M. Akhouri, Dr.(Md) Aftab A. Ansari and Dr. K. Saurav; Dr. Raghav Sharan (Clinic), Ranchi for their cooperation. RM thankful to Prof. Ajay Rana (Division of Surgical Oncology, Department of Surgery, College of Medicine, The University of Illinois at Chicago, IL 60612 USA) for providing many reagents and thanks M.Sc. students Gargee, Pratima, Swati and Priti. The fellowship MA (Project JRF), TK (CUJ Fellowship), KKP (JRF-CSIR) and AKS (DBT-RA) is acknowledged.

Authors’ contributions

MA, TK, KKP, AKS carried out the experiments. MA, RM, SN, analyzed the results. RM has written and finalized the MS and the final version of the MS has been approved by all the authors.

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Authors and Affiliations

  1. Centre for Life Sciences, School of Natural Sciences, Central University of Jharkhand, Ratu-Lohardaga Road, Brambe Ranchi, Jharkhand, 835205, India

    Manzar Alam, Tanushree Kashyap, Kamdeo K. Pramanik, Abhay K. Singh & Rajakishore Mishra

  2. Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar, Tamil Nadu, 608 002, India

    Siddavaram Nagini

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  1. Manzar Alam
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Corresponding author

Correspondence to Rajakishore Mishra.

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Conflict of interest

The authors declare that they have no conflict of interest.

Funding

The work was supported by Department of Biotechnology (DBT), New Delhi, Project No. BT/PR4624/MED/30/701/2012. Centre for Life Sciences, CUJ is supported by DBT Builder Programme No. BT/PR9028/INF/22/193/2013.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

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No role except financial assistance.

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Alam, M., Kashyap, T., Pramanik, K.K. et al. The elevated activation of NFκB and AP-1 is correlated with differential regulation of Bcl-2 and associated with oral squamous cell carcinoma progression and resistance. Clin Oral Invest 21, 2721–2731 (2017). https://doi.org/10.1007/s00784-017-2074-6

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