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Piperazine clubbed with 2-azetidinone derivatives suppresses proliferation, migration and induces apoptosis in human cervical cancer HeLa cells through oxidative stress mediated intrinsic mitochondrial pathway

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Abstract

Piperazine scaffolds or 2-azetidinone pharmacophores have been reported to show anti-cancer activities and apoptosis induction in different types of cancer cells. However, the mechanistic studies involve in induction of apoptosis addressing these two moieties for human cervical cancer cells remain uncertain. The present study emphasizes on the anti-proliferating properties and mechanism involved in induction of apoptosis for these structurally related azoles derivatives in HeLa cancer cells. 1-Phenylpiperazine clubbed with 2-azetidione derivatives (5a–5h) were synthesized, characterized using various spectroscopic techniques and evaluated for their in-vitro anti-proliferative activities and induction of apoptosis. Further, we also evaluated oxidative stress generated by these synthetic derivatives (5a–5h). Cell viability studies revealed that among all, the compound N-(3-chloro-2-(3-nitrophenyl)-4-oxoazetidin-1-yl)-2-(4-phenylpiperazin-1-yl) acetamide 5e remarkably inhibited the growth of HeLa cells in a concentration dependent manner having IC50 value of 29.44 ± 1.46 µg/ml. Morphological changes, colonies suppression and inhibition of migration clearly showed the antineoplasicity in HeLa cells treated with 5e. Simultaneously, phosphatidylserine externalization, DNA fragmentation and cell-cycle arrest showed ongoing apoptosis in the HeLa cancer cells induced by compound 5e in concentration dependent manner. Additionally, generation of intracellular ROS along with the decrease in mitochondrial membrane potential supported that compound 5e caused oxidative stress resulting in apoptosis through mitochondria mediated pathway. Elevation in the level of cytochrome c and upregulation in expression of caspase-3 clearly indicated the involvement of the intrinsic pathway of programmed cell death. In brief; compound 5e could serve as a promising lead for the development of an effective antitumor agent.

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Abbreviations

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2.5-diphenyl-tetrazolium bromide

DCFH-DA:

2′,7′-Dichlorofluorescin diacetate

AO:

Acridine organe

ROS:

Reactive oxygen species

MMP:

Mitochondrial membrane potential

RPMI:

Roswell Park Memorial Institute medium

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Acknowledgements

The research work has been funded by UGC (F no-43-172/2014 (SR)).

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Authors and Affiliations

  1. Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi, 110025, India

    Rashmin Khanam, Iram Iqbal Hejazi & Fareeda Athar

  2. Radiation and Cancer Therapeutic Lab, School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India

    Raj Kumar, Ramovatar Meena, Vikrant Jayant & Prabhat Kumar

  3. Research Centre for Nano-Materials and Energy Technology (RCNMET), School of Science and Technology, Sunway University, 47500, Selangor, Malaysia

    Syed Shahabuddin

  4. Department of Chemistry, Sri Pratap College, Cluster University, Srinagar, 190001, India

    Abdul Roouf Bhat

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  1. Rashmin Khanam
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Correspondence to Abdul Roouf Bhat or Fareeda Athar.

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10495_2018_1439_MOESM1_ESM.docx

Includes NMR spectral analysis and bar graph for cell viability assay (Figs. S.1, S.2, S.3, S.4 and S.5). (DOCX 7879 KB)

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Khanam, R., Kumar, R., Hejazi, I.I. et al. Piperazine clubbed with 2-azetidinone derivatives suppresses proliferation, migration and induces apoptosis in human cervical cancer HeLa cells through oxidative stress mediated intrinsic mitochondrial pathway. Apoptosis 23, 113–131 (2018). https://doi.org/10.1007/s10495-018-1439-x

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