In Vitro Cytotoxic Activity of Clinacanthus nutans Leaf Extracts Against HeLa Cells

Nor Hasyimah Haron,Mohammad Razak Hamdan,Nizuwan Azman,Zaleha Md Toha,Melati Khairuddean,Rafedah Abas,Hasni Arsad

doi:10.31557/apjcp.2019.20.2.601

Nor Hasyimah Haron, Mohammad Razak Hamdan + Show 5 more

Open Access

https://doi.org/10.31557/apjcp.2019.20.2.601

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Abstract

Objective:This study was conducted to investigate the antiproliferative activity of extracts of Clinacanthus nutans leaves against human cervical cancer (HeLa) cells. Methods: C. nutans leaves were subjected to extraction using 80% methanol or water. The methanol extract was further extracted to obtain hexane, dichloromethane (DCM), and aqueous fractions. The antiproliferative activity of the extracts against HeLa cells was determined. The most cytotoxic extract was furthered analyzed by apoptosis and cell cycle assays, and the phytochemical constituents were screened by gas chromatography-mass spectrometry (GC-MS). Results:All of the extracts were antiproliferative against HeLa cells, and the DCM fraction had the lowest IC50 value of 70 µg/mL at 48 h. Microscopic studies showed that HeLa cells exposed to the DCM fraction exhibited marked morphological features of apoptosis. The flow cytometry study also confirmed that the DCM fraction induced apoptosis in HeLa cells, with cell cycle arrest at the S phase. GC-MS analysis revealed the presence of at least 28 compounds in the DCM fraction, most of which were fatty acids. Conclusion:The DCM fraction obtained using the extraction method described herein had a lower IC50 value than those reported in previous studies that characterized the anticancer activity of C. nutans against HeLa cells.

Highlights

The second leading cause of death in the world is cancer, and it is estimated that cancer will contribute to 9.6 million deaths in 2018 (Bray et al, 2018)
None of the CN extracts tested in this study were cytotoxic to human cervical cancer (HeLa) cells as defined by the American National Cancer Institute’s (IC50: < 30 μg/mL) criteria of cytotoxic activity of crude extracts
Flow cytometry data showed that the DCM fraction at 48 h affected the progression of apoptosis in a dose-dependent manner, whereby the lowest dose (62.5 μg/mL) induced the highest percentage of early apoptosis and the highest dose (250 μg/mL) resulted in the most cells in the late apoptosis stage

Summary

Introduction

The second leading cause of death in the world is cancer, and it is estimated that cancer will contribute to 9.6 million deaths in 2018 (Bray et al, 2018). Numerous factors are involved in cancer development, including genetics, health, nutrition, personal habits, and the environment. The recommended treatment programme for this cancer involves surgery, chemotherapy, radiation, or other methods (World Health Organization, 2018). Herbal medicine has been increasingly recognized as a useful complementary and alternative therapy for patients with cancer. Numerous clinical studies have reported the positive effects of herbal medicines when taken with conventional therapeutics, in particular in terms of survival rate, the immune system, and quality of life of cancer patients (Yin et al, 2013)

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