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Cytotoxicity of withasteroids: withametelin induces cell cycle arrest at G2/M phase and mitochondria-mediated apoptosis in non-small cell lung cancer A549 cells

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Tumor Biology

Abstract

Considerable interest has been gained by withasteroids because of their structural uniqueness and wide spectrum of biological activities. However, limited systematic studies for proving their cytotoxic potential have so far been reported. Hence, an attempt was made to test the cytotoxicity of six withasteroids viz., withametelin (WM), withaphysalin D, withaphysalin E, 12-deoxywithastramonolide, Withaperuvin B, and physalolactone against A549, HT-29, and MDA-MB-231 cancer cell lines. Significant cytotoxic effect of WM against A549 cells (IC50 value of 6.0 μM), MDA-MB-231 cells (IC50 value of 7.6 μM), and HT-29 cells (IC50 value of 8.2 μM) was observed. Withaperuvin B and physalolactone were found to be effective against MDA-MB-231 cells. The significantly active WM arrested the A549 cells at G2/M phase and downregulated the expression of G2/M regulatory proteins such as cdc2, cyclin B1, and cdc25C. Apoptosis induced by WM in A549 cells was associated with the generation of ROS and depletion of MMP. Furthermore, WM treatment resulted in Bax upregulation, Bcl-2 downregulation, translocation of cytochrome c to mitochondria, activation of caspase-9 and −3, and PARP cleavage corroborating the apoptosis induction through intrinsic apoptotic pathway. Thus, WM possessing broader cytotoxic effect is a promising lead molecule which has the potential to be developed as a new therapeutic agent for NSCLC.

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Abbreviations

MTT:

3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide

NAC:

N-acetyl-L-cysteine

ROS:

reactive oxygen species

IC50 :

50 % inhibition of cell growth

DCFDA:

2′,7′-dichlorodihydrofluorescein diacetate

MMP:

mitochondrial membrane potential

cyt c:

cytochrome c

NSCLC:

Non-small cell lung cancer

FBS:

fetal bovine serum

PI:

Propidium iodide

PARP:

poly ADP ribose polymerase

DMSO:

dimethyl sulfoxide

RH-123:

Rhodamine 123

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Acknowledgments

Poorna Chandra Rao thanks BITS-Pilani Hyderabad Campus for granting Institute Research Fellowship (F. No. 2011PHXF420H). The authors thank Dr. Srikant Viswanadha, Incozen Therapeutics Pvt. Ltd., Hyderabad, India for extending laboratory facilities. Also, financial assistance through Research Initiation Grant from BITS-Pilani Hyderabad Campus, Telangana State, India is acknowledged.

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Authors and Affiliations

  1. Department of Pharmacy, Birla Institute of Technology and Science - Pilani Hyderabad Campus, Jawahar Nagar, Shameerpet Mandal, Hyderabad, Telangana State, 500078, India

    Poorna Chandra Rao & Sajeli Begum

  2. Medicinal Plants Processing Research Center, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran

    Mohammad Ali Farboodniay Jahromi

  3. Biology Division, Shahed 15 Education Center, Shiraz, Iran

    Zahra Hosseini Jahromi

  4. Biological Research Department, Incozen Therapeutics Pvt Ltd, Alexandria Knowledge Park, Turakapally, Hyderabad, Telangana State, India

    Saketh Sriram

  5. Department of Medicinal Chemistry, Faculty of Ayurveda, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005, India

    Mahendra Sahai

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  1. Poorna Chandra Rao
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  2. Sajeli Begum
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Correspondence to Sajeli Begum.

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Rao, P.C., Begum, S., Jahromi, M.A.F. et al. Cytotoxicity of withasteroids: withametelin induces cell cycle arrest at G2/M phase and mitochondria-mediated apoptosis in non-small cell lung cancer A549 cells. Tumor Biol. 37, 12579–12587 (2016). https://doi.org/10.1007/s13277-016-5128-5

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  • DOI: https://doi.org/10.1007/s13277-016-5128-5

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