Sains Malaysiana 48(9)(2019): 1997–2006

http://dx.doi.org/10.17576/jsm-2019-4809-21

Semi-purified Fraction of Clinacanthus nutans Induced Apoptosis in Human Cervical Cancer, SiHa Cells via Up-regulation of Bax and Down-regulation of Bcl-2

(Fraksi Separa Tulen Clinacanthus nutans Teraruh Apoptosis pada Kanser Pangkal Rahim Manusia, Sel-sel SiHa melalui Kawal Atur-atas Bax dan Kawal Atur-bawah Bcl-2)

NIK AINA SYAZANA NIK ZAINUDDIN¹, HUSSIN MUHAMMAD², NIK FAKHURUDDIN NIK HASSAN¹, NOR HAYATI OTHMAN³ & YUSMAZURA ZAKARIA¹*

¹School of Health Sciences, USM Health Campus, 16150 Kota Bharu, Kelantan Darul Naim, Malaysia

²Herbal Medicine Research Centre, Institute for Medical Research (IMR), 50588 Kuala Lumpur, Federal Territory, Malaysia

³School of Medical Sciences, USM Health Campus, 16150 Kota Bharu, Kelantan Darul Naim, Malaysia

Diserahkan: 29 Oktober 2018/Diterima: 22 Julai 2019

ABSTRACT

Cervical cancer is a leading cause of death in women, worldwide, and second in Malaysia. Nowadays, modern cancer treatment comes with negative and adverse side effects to the patients. Traditional medicinal plants such as Clinacanthus nutans has been locally recognized for its medicinal properties and was claimed for cancer treatment. According to our pilot study, a fraction of C. nutans has shown a potent inhibition on human cervical cancer cells, SiHa, in vitro. Hence, this study aimed to investigate the underlying anticancer mechanism of a semi-purified fraction (SF1) of C. nutans towards cervical cancer cells. Bioassay guided fractionation and further purification was carried out. All collected fractions of C. nutans were examined for cytotoxicity activity towards HeLa, SiHa and NIH cells by MTT Assay. SF1 exhibited better cytotoxic activity with best growth inhibition against SiHa cells (IC50 value = 9.60 μg/mL). Cytoselective effects were shown as no cytotoxicity observed in the treated normal NIH cells. SF1 induced early apoptosis event in SiHa cells as resulted by FITC-Annexin V/propidium iodide staining by flowcytometric analysis. The protein expression by flowcytometry using specific antibody conjugated fluorescent dye showed up-regulation of Bax and down-regulation Bcl-2 as well as increment of cytochrome C levels upon treatment with SF1. In conclusion, the findings suggested that SF1 demonstrated antiproliferative effects and induced apoptosis in SiHa cells through up-regulation of Bax and down-regulation of Bcl-2 via mitochondrial-dependent pathway.

Keywords: Apoptosis; cervical cancer; Clinacanthus nutans

ABSTRAK

Kanser pangkal rahim adalah penyebab utama kematian wanita di seluruh dunia dan kedua di Malaysia. Pada masa ini, rawatan moden kanser banyak memberi kesan negatif kepada pesakit. Tumbuhan ubatan tradisi seperti Clinacanthus nutans telah dikenal pasti dan dipercayai oleh penduduk tempatan sebagai alternatif kepada rawatan kanser. Menurut kajian awal kami, satu fraksi daripada C. nutans telah menunjukkan perencatan yang aktif terhadap kanser serviks manusia, secara in vitro. Oleh itu, penyelidikan ini bertujuan untuk mengkaji mekanisme antikanser oleh fraksi separa - penulenan, SF1 daripada C. nutans terhadap kanser serviks. Pemfraksian berpandukan bioaktiviti dan penulenan lanjutan telah dijalankan. Semua fraksi yang terkumpul daripada C. nutans telah diuji untuk aktiviti kesitoksikan terhadap sel HeLa, SiHa dan NIH oleh asai MTT. SF1 menunjukkan aktiviti sitotoksik yang lebih baik dengan antiproliferasi paling berpotensi terhadap sel SiHa (nilai IC50 = 9.60 μg/mL) dan menunjukkan kesan sitomemilih terhadap sel normal, NIH. Kesan apoptosis awal dapat diaruhkan terhadap sel SiHa daripada rawatan SF1 melalui analisis flositometri oleh pewarnaan FITC-Annexin V/propidium iodida. Pengekspresan protein menggunakan antibodi khusus berkonjugasi pendarfluor oleh analisis flositometri juga menunjukkan pengawalaturan Bax dan Bcl-2 serta peningkatan kadar sitokrom C setelah dirawat dengan SF1. Kesimpulannya, SF1 menunjukkan kesan antiproliferatif dan apoptosis yang diaruhkan dalam sel SiHa melalui pengawalaturan Bax dan Bcl-2.

Kata kunci: Apoptosis; Clinacanthus nutans; kanser serviks

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*Pengarang untuk surat-menyurat; email: yusmazura@usm.my