Open AccessTRIGONELLA FOENUM-GRAECUM L. EXHIBITS ESTROGENIC EFFECT THROUGH PRESENILIN 2 GENE EXPRESSION IN THE BREAST CANCER CELL LINE MCF-7
Author(s) -
Michael Wink,
Wahono Sumaryono,
Fransiscus D. Suyatna,
Nurjati Chairani Siregar
Publication year - 2019
Publication title -
asian journal of pharmaceutical and clinical research
Language(s) - English
Resource type - Journals
eISSN - 2455-3891
pISSN - 0974-2441
DOI - 10.22159/ajpcr.2019.v12i1.29340
Subject(s) - mcf 7 , trigonella , gene expression , ethyl acetate , cytotoxic t cell , chemistry , microbiology and biotechnology , housekeeping gene , cell culture , western blot , biology , gene , biochemistry , cancer cell , in vitro , cancer , botany , genetics , human breast
Objective: The objective of this study is to investigate the estrogenic and antiestrogenic activity of Fenugreek seeds, Trigonella foenum-graecum L. in the estrogen-dependent breast cancer cell line, MCF-7, including its effect on the expression of estrogen-dependent presenilin 2 (pS2) gene.Methods: An activity guided fractionation was carried out with extracts from fenugreek seeds in MCF-7 cells. Cytotoxic activity assays were conducted with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. Most fractions were also tested also tested in media with estradiol 10 nM We also analysed the expression of pS2 gene. For the analysis of pS2 gene expression we employed PCR primers for pS2 and for β-actin as a housekeeping gene using real-time polymerase chain reaction (RT-PCR).Results: Based on cytotoxic activity assay in MCF-7, the active fractions are ethyl acetic fraction and its phases ethyl acetic (EA) 2 and EA 2.2. The most active fraction was EA 2.2 (IC50=27.129 ppm), which exhibited a biphasic effect; at low concentrations, it stimulated the growth, and at high concentrations it showed strong cytotoxic effects. EA2.2 fraction in concentration 20 ppm, also could induce pS2 gene expression in media with and without estrogen.Conclusion: The most active fraction was the ethyl acetate phase and further subfractions. The most active fraction also induced the expression of pS2 gene which was studied by RT-PCR.