Abstract
Sinigrin, a precursor of allyl isothiocyanate, present in the Raphanus sativus exhibits diverse biological activities, and has an immense role against cancer proliferation. Therefore, the objective of this study was to quantify the sinigrin in the R. sativus roots using developed and validated RP-HPLC method and further evaluated its’ anticancer activity. To achieve the objective, the roots of R. sativus were lyophilized to obtain a stable powder, which were extracted and passed through an ion-exchange column to obtain sinigrin-rich fraction. The RP-HPLC method using C18 analytical column was used for chromatographic separation and quantification of sinigrin in the prepared fraction, which was attained using the mobile phase consisting of 20 mM tetrabutylammonium: acetonitrile (80:20%, v/v at pH 7.0) at a flow rate of 0.5 mL/min. The chromatographic peak for sinigrin was showed at 3.592 min for pure sinigrin, where a good linearity was achieved within the concentration range of 50 to 800 µg/mL (R2 > 0.99), with an excellent accuracy (−1.37% and −1.29%) and precision (1.43% and 0.94%), for intra and inter-day, respectively. Finally, the MTT assay was performed for the sinigrin-rich fraction using three different human cancer cell lines, viz. prostate cancer (DU-145), colon adenocarcinoma (HCT-15), and melanoma (A-375). The cell-based assays were extended to conduct apoptotic and caspase-3 activities, to determine the mechanism of action of sinigrin in the treatment of cancer. MTT assay showed IC50 values of 15.88, 21.42, and 24.58 µg/mL for DU-145, HCT-15, and A-375 cell lines, respectively. Increased cellular apoptosis and caspase-3 expression were observed with sinigrin-rich fraction, indicating significant increase in overexpression of caspase-3 in DU-145 cells. In conclusion, a simple, sensitive, fast, and accurate RP-HPLC method was developed for the estimation of sinigrin in the prepared fraction. The data observed here indicate that sinigrin can be beneficial in treating prostate cancer possibly by inducing apoptosis.
Highlights
Accumulation of active phytoconstituents within the different parts of plants are nowadays explored extensively to use in the ailments of several diseases; we are re-entering into the era of exploring and using medicinal plants
7 of 16 of this study indicated that the tested samples induce apoptosis to the experimental cell line
Our results showed that R. sativus roots extract and sinigrin showed a cytotoxic effect against all three cell lines and restrained the growth of the cells in a dose-dependent manner
Summary
Accumulation of active phytoconstituents within the different parts of plants are nowadays explored extensively to use in the ailments of several diseases; we are re-entering into the era of exploring and using medicinal plants. Thereby, we encounter tremendous usage of the new buzzword ‘phytochemicals’ in recent research These studies involved looking for isolated active chemicals within plants instead of using the whole plants as performed by our ancestors. Various epidemiological studies in the last few decades have demonstrated the importance of consumption of diet rich in fruits and vegetables to reduce the incidence of tumor formation as well as other cardiovascular diseases [3,4,5]. In this perspective, the brassicaceous vegetables are rich in phytochemicals and have demonstrated their nutritional and health benefits [6,7]. Radish contains some unique bioactive chemical constituents, glucosinolates, a large group of anionic sulfur-containing compounds, which exhibited various biological activities and could be utilized in curing different human diseases [13,14]
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