Optimization Sweet Potato [Ipomoea Batats (L.) Lam] Sporamin Extraction and Analyzing its Antiproliferative Effect on Breast Cancer Cells, MCF-7 Cell Line

Abstract

Background: Finding cures with herbal origins with an easy and cost-effective way of extraction for treatment of diseases such as cancer is highly appreciated. Considering that proteases are involved in initiation and spreading of cancer cells, proteases inhibitors can be take into account as an option for treatment. Sporamin is belonging to trypsine inhibitors group which exists in sweet potato tuber. In this study, the method of sporamin extracting was optimized, and for the first time, its effect on proliferation of breast cancer, MCF-7 cell line, was examined. Methods: Aqueous extraction was obtained from sweet potato. Transparent extraction was inserted into chromatography diethylaminoethanol-Sepharose (DEAE-Sepharose) column. After washing with a linear salt gradient, fractions were separated. To determine protein purity, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was performed. Sporamin inhibitory activity was measured using Laskowaski method, and eventually 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT method) was used to assess the antiproliferative effect of sporamin. Findings: A single band of 25 kDa sporamin was obtained via SDS-PAGE. Its inhibitory effect was measured 800 units in mg/minute. According to the results of MTT method, the amounts of inhibitory concentration 50 (IC50) for 24 and 48 hours were significantly different (P < 0.01). Conclusion: Using single-column ion exchange and linear salt gradient for sporamin purification was successful; and due to its antiproliferative effect on MCF-7 cells, there is hope that this technique could be used for the purification of drug and treatment of disease in future.

Keywords

• Ipomoea batatas
• Purification
• Chromatography
• Electrophoresis
• Polyacrylamide gel
• Breast neoplasms