Sains Malaysiana 52(6)(2023): 1811-1820

http://doi.org/10.17576/jsm-2023-5206-16

The Effect of Lunasin on Inhibition of KI67, BCL-2 and C-MYC Expression in Azoxymethane and Dextran Sodium Sulfate Induced Mice Colon  

(Kesan Lunasin terhadap Perencatan Ekspresi KI67, BCL-2 dan C-MYC dalam Kolon Tikus Terinduksi Azoksimetana dan Dektran Natrium Sulfat)

KUSMARDI KUSMARDI^1,2,3,4,*, AHMAT REDIANSYA PUTRA⁵ & RIA KODARIAH¹

¹Department of Anatomical pathology, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia

²Drug Development Research Cluster, Indonesian Medical Education and Research Institute, Universitas Indonesia, Jakarta, Indonesia

³Human Cancer Research Cluster, Indonesian Medical Education and Research Institute, Universitas Indonesia, Jakarta, Indonesia

⁴Doctoral Program for Biomedical Sciences, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia

⁵Graduate Student, Master’s Program in Biomedical Science, Faculty of Medicine, Universitas Indonesia, Jalan Salemba Raya No 6, Jakarta, Indonesia

Diserahkan: 26 Julai 2022/Diterima: 2 Jun 2023

Abstract

Treatment of cancer using medicinal-plant based has been important due to minimal side effects, high efficiency and low cost. Lunasin from soybean is known as potential chemopreventive agent. This study aimed to study and investigate the proteins involved in the mechanisms of action of lunasin underlie its chemopreventive effects in Azoxymethane (AOM) and Dextran Sodium Sulfate (DSS) induced mice. A total 30 BLAB/c mice were separated into six groups. In five of the groups - a negative control group, positive control group, and three intervention groups - carcinogenesis was induced with AOM and DSS; the sixth group received no interventions. Lunasin were given in different doses of Low Dose Lunasin (75 mg/kgBW), Moderate Dose Lunasin (150 mg/kgBW), and High Dose Lunasin (200 mg/kgBW) to intervention groups. Immunohistochemistry was conducted to measure Ki67, C-myc, and Bcl-2 expressions from the distal colons of mice that had been sacrificed. The samples were microscopically assessed and photographed, and cell counts were performed using the Image J application. Further, the H-score method was used to quantify of Ki67, C-myc and Bcl-2 expressions. The results of this show that there is significant differences between the negative control and the intervention groups were found at the 75 mg/kgBW and 150 mg/kgBW (p < 0.05) lunasin dosage levels. This demonstrates that Lunasin inhibits proliferation and induces apoptosis in the colon mice induced by AOM and DSS.

Keywords: AOM; Bcl-2; C-myc; DSS; Ki67

Abstrak

Rawatan kanser berasaskan tumbuhan ubatan adalah penting kerana kesan sampingan yang minimum, kecekapan tinggi dan kos yang rendah. Lunasin daripada kacang soya dikenali sebagai agen kemohalang yang berpotensi. Penyelidikan ini bertujuan untuk mengkaji protein yang terlibat dalam mekanisme tindakan lunasin yang mendasari kesan kemohalangnya pada tikus yang disebabkan oleh Azoksimetana (AOM) dan Dektran Natrium Sulfat (DSS). Sebanyak 30 tikus BLAB/c dipisahkan kepada enam kumpulan. Dalam lima daripada kumpulan kawalan negatif, kumpulan kawalan positif dan tiga kumpulan intervensi-karsinogenesis telah diinduksi dengan AOM dan DSS; kumpulan keenam tidak menerima campur tangan. Lunasin diberikan dalam dos berbeza Lunasin Dos Rendah (75 mg/kgBW), Lunasin Dos Sederhana (150 mg/kgBW) dan Lunasin Dos Tinggi (200 mg/kgBW) kepada kumpulan intervensi. Imunohistokimia telah dijalankan untuk mengukur ekspresi Ki67, C-myc dan Bcl-2 daripada kolon distal tikus yang telah dikorbankan. Sampel telah dinilai secara mikroskopik dan difoto dan kiraan sel dilakukan menggunakan aplikasi Image J. Selanjutnya, kaedah H-skor digunakan untuk mengukur ungkapan Ki67, C-myc dan Bcl-2. Keputusan ini menunjukkan terdapat perbezaan yang signifikan antara kawalan negatif dan kumpulan intervensi didapati pada tahap dos lunasin 75 mg/kgBW dan 150 mg/kgBW (p < 0.05). Ini menunjukkan bahawa Lunasin menghalang percambahan dan mendorong apoptosis dalam tikus kolon yang disebabkan oleh AOM dan DSS.

Kata kunci: AOM; Bcl-2; C-myc; DSS; Ki67

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*Pengarang untuk surat-menyurat; email: kusmardis@gmail.com