IMMUNOGENICITY ANALYSIS OF TRITERPENE GLYCOSIDE FROM HOLOTHURIA ATRA TO DETECTING FAS AND BCL-2 PROTEIN ON THE SP-C1 CELL OF TONGUE CARCINOMA

Utmi Arma; Mieke Hemiawati Satari; Syafruddin Ilyas; Dian Handayani; Ameta Primasari; Nila Kasuma; Supriatno; Basri A Gani

doi:10.22159/ajpcr.2018.v11i6.25065

Authors

Utmi Arma Doctoral Program, Dentistry Faculty, Universitas Sumatera Utara, Medan, Indonesia.
Mieke Hemiawati Satari Department of Microbiology, Dentistry Faculty, Universitas Padjajaran, Bandung, Indoensia.
Syafruddin Ilyas Department of Biology, Mathematics and Natural Sciences Faculty, Universitas Sumatera Utara, Medan, Indonesia.
Dian Handayani Department of Pharmacy, Pharmacy Faculty, Universitas Andalas, Padang, Indoensia.
Ameta Primasari Department of Oral Biology, Dentistry Faculty, Universitas Sumatera Utara, Medan, Indonesia.
Nila Kasuma Department of Oral Biology, Dentistry Faculty, Universitas Andalas, Padang, Indonesia.
Supriatno Department of Oral Medicine, Dentistry Faculty, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Basri A Gani Department of Oral Biology, Dentistry Faculty, Universitas Syiah Kuala, Banda Aceh, Indonesia.

DOI:

https://doi.org/10.22159/ajpcr.2018.v11i6.25065

Keywords:

Holothuria atra, Cytotoxic-apoptosis, Bcl-2 and Fas proteins, Nil, Triterpene glycoside

Abstract

Objective: The objective of this study is to assess the role of triterpene glycoside of Holothuria atra to induce the Fas and Bcl-2-regulated apoptosis in Supri's Clone 1 (Sp-C1) cell of tongue carcinoma.

Methods: The triterpene glycoside of H. atra was isolated by high-performance liquid chromatography. The Sp-C1 cell of tongue carcinoma was cloned by Dulbecco's Modified Eagle Medium and cytotoxicity assay by 3-4-5-dimethylthiazol-2yl 2,5-diphenyltetrazolium bromide assay. Expression Fas and Bcl-2 protein were analysed by immunocytochemistry also apoptosis detected by double staining ethidium bromide acridine. The datum of studied was analyzed by one-way analysis of variance (ANOVA), significance (p<0.05), and strength correlation (p<0.001) with R=1.

Results: The H. atra has triterpene glycoside, and in the dose of 4 mg/ml, it has been cytotoxic activities on the Sp-C1 (p<0.05), mortality 80%; inhibitory concentration 50 (IC50)=0.6 and anti-logarithm = 4. In general, the concentration of 2.5 mg/ml of triterpene glycoside has triggered the expression of Fas protein (active, 71%; moderate, 10%; and no-active, 27%), whereas the Bcl-2 protein (active, 59%; moderate, 14%; and no-active 27%). Statistically, both expressions of protein were significant (p<0.05). Triterpene glycoside caused the apoptosis of Sp-C1 cell (strong, 87%, and moderate, 13%).

Conclusion: The triterpene glycoside has the properties of cytotoxicity, and apoptosis in the SP-C1 cell also could be triggering the expression of Fas and Bcl-2 proteins.

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Author Biography

Utmi Arma, Doctoral Program, Dentistry Faculty, Universitas Sumatera Utara, Medan, Indonesia.

Oral Biology Department

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