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Hedychium coronarium extract arrests cell cycle progression, induces apoptosis, and impairs migration and invasion in HeLa cervical cancer cells

Authors Ray A, Jena S, Dash B, Sahoo A, Kar B, Patnaik J, Panda PC, Nayak S, Mahapatra N

Received 7 October 2018

Accepted for publication 28 November 2018

Published 3 January 2019 Volume 2019:11 Pages 483—500


Checked for plagiarism Yes

Review by Single-blind

Peer reviewer comments 2

Editor who approved publication: Dr Xueqiong Zhu

Asit Ray,1,2 Sudipta Jena,2 Biswabhusan Dash,2 Ambika Sahoo,2 Basudeba Kar,2 Jeetendranath Patnaik,3 Pratap Chandra Panda,4 Sanghamitra Nayak,2 Namita Mahapatra1

1Regional Medical Research Centre (Indian Council of Medical Research), Chandrasekharpur, Bhubaneswar 751023, Odisha, India; 2Centre for Biotechnology, School of Pharmaceutical Sciences, Siksha O Anusandhan University, Kalinganagar, Ghatikia, Bhubaneswar 751003, Odisha, India; 3Department of Botany, Sri Krushna Chandra Gajapati College, Paralakhemundi 761200, Odisha, India; 4Taxonomy and Conservation Division, Regional Plant Resource Centre, Nayapalli, Bhubaneswar 751015, Odisha, India

Background: Hedychium coronarium Koen. (Zingiberaceae) is traditionally used as medicine in countries such as India, China, and Vietnam for treatment of various ailments including cancer. However, in spite of its implied significance in cancer treatment regimes, there are no reports so far involving the anticancerous attributes of H. coronarium ethanol extract (HCEE) on cancer cells and a more comprehensive study on its mechanism is still lacking.
Materials and methods: The cytotoxicity of HCEE was evaluated by MTT and clonogenic survival assay. Annexin V/propidium iodide (PI), Hoechst 33342 staining, and TUNEL assay were performed to detect apoptosis. Cell cycle analysis was performed using PI staining. JC-1 and 2’,7’-dichlorodihydrofluorescein diacetate assay were used to check the levels of MMP and ROS, respectively. Western blot analysis was carried out to measure the expression levels of proteins. Migration and invasion activity were assessed by wound healing and Transwell membrane assay, respectively.
Results: Antiproliferative effect of HCEE was investigated in various cancerous and normal cell lines. Among these, HCEE significantly inhibited the survival of HeLa cells without affecting the viability of normal human umbilical vein endothelial cells. Annexin V/PI, Hoechst staining, and TUNEL assay showed HCEE induced apoptosis in HeLa cells in a dose-dependent manner. HCEE promoted cell cycle arrest at G1 phase in HeLa cells by upregulating the levels of p53 and p21 and downregulating the levels of cyclin D1, CDK-4, and CDK-6. Moreover, HCEE treatment upregulated the expression of Bax and downregulated the expression of Bcl-2. Additionally, HCEE activated the caspase cascade by increasing the activities of caspase-9, caspase-8, and caspase-3. The expression levels of Fas ligand and Fas were also upregulated. Further, HCEE inhibited the migratory potential of HeLa cells by downregulating MMP-2 and MMP-9 expression levels.
Conclusion: Our results indicate H. coronarium exerts antiproliferative and apoptotic effects against HeLa cells, and therefore may be used for treatment against cervical cancer.

Keywords: apoptosis, cervical cancer, cytotoxicity, Hedychium coronarium extract, HeLa cell line

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