Sains Malaysiana 53(3)(2024): 667-679

http://doi.org/10.17576/jsm-2024-5303-14

SF1: A Standardised Fraction of Clinacanthus nutansthat Inhibits the Stemness Properties of Cancer Stem-Like Cells Derived from Cervical Cancer

(SF1: Fraksi Piawaian Clinacanthus nutans yang Merencat Sifat Stem Sel Menyerupai Sel Stem Kanser Diperolehi daripada Kanser Serviks)

FARIDAH ISMAIL^1,2, YUSMAZURA ZAKARIA^1,*, MUHAMMAD LOKMAN MD ISA³, NIK FAKHURUDDIN NIK HASSAN⁴ & TAN SUAT CHENG¹

¹Biomedicine Program, School of Health Sciences, Health Campus, Universiti Sains Malaysia, Kubang Kerian, 16150 Kota Bharu, Kelantan, Malaysia

²Department of Basic Medical Sciences, Kulliyyah of Medicine, International Islamic University of Malaysia, Jalan Sultan Ahmad Shah, Bandar Indera Mahkota, 25200 Kuantan, Pahang, Malaysia

³Department of Basic Medical Sciences, Kulliyyah of Nursing, International Islamic University of Malaysia, Jalan Sultan Ahmad Shah, Bandar Indera Mahkota, 25200 Kuantan, Pahang, Malaysia

⁴Forensic Science Program, School of Health Sciences, Health Campus, Universiti Sains Malaysia, Kubang Kerian, 16150 Kota Bharu, Kelantan, Malaysia

Received: 24 July 2023/Accepted: 21 February 2024

Abstract

Cancer stem cells (CSCs) are a small population of tumour cells that are responsible for tumour initiation, metastases, recurrence, and resistance to conventional therapy. Hence, targeting CSCs is crucial in the fight against cancer. SF1, a standardised fraction from Clinacanthus nutans leaf extract, has been reported to exhibit potent and selective antineoplastic effects against cervical cancer cells. In this study, the potential of SF1 to inhibit the stemness of cervical cancer stem-like cells has been evaluated. SF1 extraction was carried out using the dry column vacuum chromatography technique. SiHa cell lines were cultured as spheres in CSC-conditioned medium (cervospheres), and the IC₅₀ of SF1 against cervospheres was determined using the OZBlue Cell Viability Kit. The effects of SF1 on the cervosphere’s stemness markers, including CD49f, CK17, SOX2, OCT4, and NANOG, were assessed using a flow cytometry assay. Self-renewal inhibition and anti-tumorigenesis effects of SF1 in cervospheres were evaluated using a sphere formation assay and a xenograft mouse model. The present study shows that SF1 treatment at an IC₅₀ of 17.07 µg/mL inhibited the proliferation, self-renewal, and tumorigenic capacity of SiHa cervospheres in vitro and in vivo. A decrease in the expressions of CK17, SOX2, CD49f, and OCT4 in cervical CSCs indicated that SF1’s inhibitory effects were also associated with the suppression of stemness markers. These results suggest that SF1 possesses an antitumor effect against cervical CSCs and may be regarded as a promising approach to the development of targeted anticancer agents for cervical cancer.

Keywords: Anticancer; cancer stem cells; cervical cancer; Clinacanthus nutans; stemness

Abstrak

Sel stem kanser adalah populasi kecil daripada keseluruhan sel tumor yang bertanggungjawab mencetus pertumbuhan awal tumor, perebakan, pengulangan dan kerintangan terhadap rawatan konvensional. Oleh itu, sasaran rawatan terhadap sel stem kanser adalah penting bagi memerangi kanser. SF1, fraksi yang diseragamkan daripada ekstrak daun Clinacanthus nutans, dilaporkan menunjukkan kesan anti-kanser yang kuat dan selektif terhadap sel kanser serviks. Dalam kajian ini, potensi SF1 untuk merencatkan ciri stem sel yang menyerupai sel stem kanser serviks telah dikaji. SF1 diekstrak menggunakan teknik kromatografi vakum lajur kering. Sel SiHa telah dikultur sebagai sel sfera di dalam kultur media khusus untuk pembentukan sel stem kanser (sel sfera kanser serviks) dan nilai IC₅₀ SF1 terhadap sel sfera kanser serviks ditentukan menggunakan kit kelangsungan hidup OZBlue. Kesan SF1 pada penanda stem di dalam sel sfera kanser serviks, termasuk CD49f, CK17, SOX2, OCT4 dan NANOG, dianalisis menggunakan ujian flositometri. Perencatan pembaharuan diri dan kesan anti-tumorigenesis SF1 dalam sel sfera kanser serviks dinilai menggunakan ujian pembentukan sel sfera dan model tetikus xenocantum. Kajian ini menunjukkan bahawa rawatan SF1 pada IC₅₀ 17.07 μg/mL merencat proliferasi, pembaharuan diri dan keupayaan tumorigenik sel sfera kanser serviks SiHa in vitro dan in vivo. Kesan perencatan SF1 turut disertai oleh penurunan penanda stem untuk sel stem kanser serviks, seperti yang ditunjukkan oleh penurunan dalam pengekspresan CK17, SOX2, CD49f dan OCT4. Hasil kajian ini menunjukkan bahawa SF1 mempunyai kesan anti-tumor terhadap sel stem kanser serviks dan berpotensi sebagai pendekatan baharu untuk membangunkan agen anti-kanser yang bersasar terhadap kanser serviks.

Kata kunci: Anti-kanser; Clinacanthus nutans; kanser serviks; sel stem kanser; sifat stem

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*Corresponding author; email: yusmazura@usm.my