Sains Malaysiana 44(8)(2015): 1137–1143

Aloe Emodin Induces Apoptosis in ER⁺-breast Cancer Cells; MCF-7 through IGF-1R Signalling Pathway

(Aloe Emodin Mengaruh Apoptosis dalam Sel Kanser Payu Dara ER⁺; MCF-7 melalui Tapak Jalan Pengisyaratan IGF-1R)

INDAH MOHD AMIN^1,5, ROZIANA KAMALUDIN¹, SWEE KEONG YEAP⁷, MOHAMAD

RODI ISA⁴, NIK MOHD MAZUAN NIK MOHD ROSDY⁶, ROSFAIIZAH SIRAN³, SITI HAMIMAH SHEIKH ABDUL KADIR^1,2 & NARIMAH ABDUL HAMID HASANI²*

¹Institute for Medical Molecular Biotechnology (IMMB), Faculty of Medicine, Sungai Buloh Campus

Universiti Teknologi MARA (UiTM), Jalan Hospital, 47000 Sungai Buloh, Selangor Darul Ehsan

Malaysia

²Department of Biochemistry and Molecular Medicine, Faculty of Medicine, Sungai Buloh Campus

Universiti Teknologi MARA (UiTM), Jalan Hospital, 47000 Sungai Buloh, Selangor Darul Ehsan

Malaysia

³Department of Physiology, Faculty of Medicine, Sungai Buloh Campus, Universiti Teknologi MARA (UiTM), Jalan Hospital, 47000 Sungai Buloh, Selangor Darul Ehsan, Malaysia

⁴Population Health and Preventive Medicine, Faculty of Medicine, Sungai Buloh Campus

Universiti Teknologi MARA (UiTM), Jalan Hospital, 47000 Sungai Buloh, Selangor Darul Ehsan

Malaysia

⁵Centre of Preclinical Sciences Studies, Faculty of Dentistry, Sungai Buloh Campus, Universiti Teknologi MARA (UiTM), Jalan Hospital, 47000 Sungai Buloh, Selangor Darul Ehsan

Malaysia

⁶Centre of Oral and Maxillofacial Diagnostic and Medicine Studies, Faculty of Dentistry,

Sungai Buloh Campus, Universiti Teknologi MARA (UiTM), Jalan Hospital, 47000 Sungai Buloh, Selangor Darul Ehsan, Malaysia

⁷Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan

Malaysia

Diserahkan: 16 September 2014/Diterima: 6 April 2015

ABSTRACT

Two-third of breast cancer patients expressed estrogen receptors (ER)s and received endocrine treatment with established anti-estrogens such as tamoxifen. But the action and acquired resistance during treatment are largely unknown. In contrary, phytochemicals are more selective and less cytotoxic to normal cells. Accordingly, we found aloe emodin, an anthraquinone to inhibit the proliferation of ER⁺-breast cancer cells, MCF-7 with IC₅₀ of 80μM, but not affecting control breast cells, MCF-10A. Tamoxifen was non-selective to both cells with IC₅₀ of 27 μM and 38 μM, respectively. Thus, we aimed to investigate the anti-proliferative mechanism of aloe emodin on MCF-7 and its underlying signalling compared to tamoxifen. Cells were treated separately with aloe emodin and tamoxifen at respective IC₅₀ for 72 h. Apoptosis was determined using Annexin V-FITC/PI staining. The expression of insulin-like growth factor-1 receptor (IGF-1R), insulin-like growth factor binding protein (IGFBP)-2 and B-raf gene was investigated using QuantiGene 2.0 Plex assay. Paired-student t-test and ANOVA test were used to compare between untreated and treated cells on the measured parameters. Each treatment was conducted in triplicate and repeated three times. Significance was set at p<0.05. The presences of early and late apoptosis in MCF-7 were seen in both treatments. All target genes were down regulated. The anti-proliferation effect of aloe emodin on MCF-7 is similar with tamoxifen which mediates inhibition of IGF-1R signalling pathway. This suggests aloe emodin as a potential anti-cancer agent to be used in combined anti-estrogen therapy to enhance its efficacy in ER+-breast cancer treatment.

Keywords: Aloe emodin; apoptosis; IGF-1R; MCF-7

ABSTRAK

Dua-pertiga pesakit kanser payu dara mengekspresi reseptor estrogen (RE) dan menerima rawatan endokrin dengan anti-estrogen seperti tamoksifen. Tetapi, tindakan dan kerentanan perolehan terhadap rawatan ini masih belum difahami. Sebaliknya, fito-kimia didapati lebih selektif dan kurang memberi kesan toksik kepada sel normal. Selanjutnya, kajian kami mendapati aloe emodin, sejenis antrakuinon berjaya merencat proliferasi sel kanser payu dara ER⁺, MCF-7 dengan IC₅₀ 80μM, tanpa memberi kesan toksik kepada sel payu dara normal, MCF-10A. Sebaliknya, tamoksifen adalah tidak selektif kepada kedua-dua sel dengan IC₅₀ masing-masing 27 dan 38 μM. Oleh itu, penyelidikan ini dijalankan untuk mengkaji mekanisme anti-proliferasi dan laluan pengisyaratan transduksi aloe emodin terhadap MCF-7 berbanding dengan tamoksifen. Sel diaruhkan dengan aloe emodin dan tamoksifen secara berasingan pada IC₅₀ selama 72 jam. Apoptosis ditentukan dengan ujian Annexin V-FITC/PI pewarnaan. Pengekspresan gen-gen faktor tumbesaran-1 reseptor (IGF-1R) seperti-insulin, faktor tumbesaran pengikat protein (IGFBP)-2 seperti-insulin dan B-raf ditentukan menggunakan asai Plex QuantiGene 2.0. Ujian-T pelajar-berpasangan dan ANOVA digunakan untuk membandingkan antara parameter sel teraruh dan kawalan. Setiap perlakuan dijalankan sebanyak tiga kali dan diulang tiga kali. Tahap signifikan ditetapkan pada p<0.05. Tanda-tanda awal dan akhir apoptosis di MCF-7 diperhatikan pada kedua-dua aruhan. Ketiga-tiga gen diekspreskan kurang daripada paras normal. Kesan anti-proliferasi aloe emodin terhadap MCF-7 adalah bersamaan dengan tamoksifen iaitu melalui perencatan tapak jalan pengisyaratan tranduksi IGF-1R. Kajian ini mencadangkan aloe emodin berpotensi sebagai agen anti-kanser yang boleh digunakan sebagai rawatan gabungan dengan terapi anti-estrogen yang sedia ada, bertujuan untuk meningkatkan efikasi rawatan tersebut.

Kata kunci: Aloe emodin; apoptosis; IGF-1R; MCF-7

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*Pengarang untuk surat-menyurat; email: drnarimah@salam.uitm.edu.my