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Benzothiazole derivative bearing amide moiety induces p53-mediated apoptosis in HPV16 positive cervical cancer cells

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In our previous study, we screened the anti-cancer properties of 10 benzothiazole derivatives in cervical cancer cell lines. In the present study, we aimed to delineate the mechanism of the apoptotic pathway (whether intrinsic or extrinsic) following the treatment of N-(4-(benzo[d]thiazol-2-yl)phenyl)-5-chloro-2-methoxybenzamide (named as A-07) on cervical cancer cell lines. Cellular stress by reactive oxygen species was measured using DCFDA dye by flowcytometry. Protein expression and localization was checked by immunofluorescence for γH2A.X, TP53, and CASP-3. Expression profiles of BAX and BCL-2 was done by semi-quantitative RT-PCR and PARP-1 (Poly(ADP-ribose) polymerase-1) by Western blot analysis. Bioinformatic studies were done using PDB websites, metaPocket 2.0 server, YASARA software and Discovery Studio 3.5 Visualizer. We demonstrate that the compound A-07 leads to ROS generation and double strand breaks in SiHa and C-33A cells. The induction of apoptosis in SiHa cells is associated with increased nuclear expression of the tumor suppressor protein, TP53. The shift in BAX/BCL-2 ratio, increased expression of Caspase-3 and cleaved Poly(ADP-ribose) polymerase-1 favour apoptotic signal in SiHa. In silico studies revealed that A-07 has inhibiting capabilities to the E6/E6AP/P53 complex. Our data suggest that treatment of A-07 causes p53 and caspase dependent apoptosis in HPV 16 infected SiHa cells.

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Acknowledgements

University Grants Commission, New Delhi and Banaras Hindu University DST-PURSE programme for Junior and Senior Research fellowships to AM; Banaras Hindu University UGC-UPE and Banaras Hindu University DST-PURSE programmes for research partial funding to GN and SS; Department of Biotechnology, Banaras Hindu University and Interdisciplinary School of Life Sciences, Banaras Hindu University for equipment facilities.

Funding

The work was supported by University Grants Commission, New Delhi and Banaras Hindu University DST-PURSE programme for Junior and Senior Research fellowships to AM; Banaras Hindu University UGC-UPE and Banaras Hindu University DST-PURSE programmes for research funding to GN and SS.

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Authors and Affiliations

  1. Department of Molecular and Human Genetics, Institute of Science, Banaras Hindu University, Varanasi, 221005, India

    Arusha Modi, Ozasvi R. Shanker & Gopeshwar Narayan

  2. Department of Pharmaceutical Engineering and Technology, Indian Institute of Technology, Banaras Hindu University, Varanasi, 221005, India

    Meenakshi Singh, Gopichand Gutti & Sushil Kumar Singh

  3. School of Biotechnology, Institute of Science, Banaras Hindu University, Varanasi, 221005, India

    Vinay Kumar Singh

  4. Department of Zoology, Mahila Mahavidyalaya, Banaras Hindu University, Varanasi, 221005, India

    Sunita Singh

  5. Department of Radiotherapy & Radiation Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh, 221 005, India

    Satyajit Pradhan

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  1. Arusha Modi
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Modi, A., Singh, M., Gutti, G. et al. Benzothiazole derivative bearing amide moiety induces p53-mediated apoptosis in HPV16 positive cervical cancer cells. Invest New Drugs 38, 934–945 (2020). https://doi.org/10.1007/s10637-019-00848-7

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