Abstract
Crocin, the main pigment of Crocus sativus L., has been shown to have antiproliferative effects on cancer cells, but the involved mechanisms are only poor understood. This study focused on probable effect of crocin on the immortality of hepatic cancer cells. Cytotoxicity of crocin (IC50 3 mg/ml) in hepatocarcinoma HepG2 cells was determined after 48 h by neutral red uptake assay and MTT test. Immortality was investigated through quantification of relative telomerase activity with a quantitative real-time PCR-based telomerase repeat amplification protocol (qTRAP). Telomerase activity in 0.5 μg protein extract of HepG2 cells treated with 3 mg/ml crocin was reduced to about 51% as compared to untreated control cells. Two mechanisms of inhibition, i.e. interaction of crocin with telomeric quadruplex sequences and down regulation of hTERT expression, were examined using FRET analysis to measure melting temperature of a synthetic telomeric oligonucleotide in the presence of crocin and quantitative real-time RT-PCR, respectively. No significant changes were observed in the Tm telomeric oligonucleotides, while the relative expression level of the catalytic subunit of telomerase (hTERT) gene showed a 60% decrease as compared to untreated control cells. In conclusion, telomerase activity of HepG2 cells decreases after treatment with crocin, which is probably caused by down-regulation of the expression of the catalytic subunit of the enzyme.
Highlights
Cancer is one of the most important health problems around the world, in older patients and in children
Telomerase activity in 0.5 μg protein extract of HepG2 cells treated with 3 mg/ml crocin was reduced to about 51% as compared to untreated control cells
Telomerase activity of HepG2 cells decreases after treatment with crocin, which is probably caused by down-regulation of the expression of the catalytic subunit of the enzyme
Summary
Cancer is one of the most important health problems around the world, in older patients and in children. Chemoprevention of cancer especially by natural compounds is a promising strategy against cancer initiation. In this regard natural compounds with strong antioxidative, hepatoprotective and anti-inflammatory effects are interesting candidates to evaluate their ability of influence the initiation and growth of tumors. Saffron, obtained from the dried stigma of Crocus sativus L., is an important spice rich in terpenoids. It is commonly consumed in different parts of the world and used as a medicinal drug to treat several health disorders. Saffron extracts were shown to inhibit papilloma growth, decline the incidence of squamous cell carcinoma and soft tissue sarcoma in mice. Crocin appears to exhibit an anticarcinogenic activity (Konoshima & Takasaki, 2003)
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