Cytotoxic effect of piper sarmentosum ROXB. methanolic extract against human breast cancer MCF-7 cell line

Abstract

Breast cancer is the most common cancer in women in almost all part of the world. Breast cancer treatment has been one of the major concerns in the world nowadays because of the rise in the number of newly reported cases and most of the anticancer drugs used are reported to have high toxicity towards normal cells. Therefore, it is crucial to discover anticancer drugs from plants natural products but with fewer side effects. Malaysia is one of the plants mega diversity countries of the world, possesses plants had been used for centuries to treat human disorders and diseases. One of them is Piper sarmentosum Roxb., locally known as kaduk and commonly used in Malaysia in cuisine and folk medicine. The plant has been used traditionally to treat many diseases, including fever, jaundice, rheumatic arthritis, neuralgia and as a carminative, fungal infections, pleurisy, asthmatic coughing and as an expectorant. It is believed to have an antifungal and antioxidant activity. The aim of this study is to determine the optimized processing conditions of ultrasound assisted extraction (UAE) of P. sarmentosum methanolic extract and also to purify and identify the compound responsible for antiproliferative activity against human breast cancer cells from the crude extract of P. sarmentosum. The ultrasonic assisted extraction was optimized using Response Surface Method (RSM) to indicate the optimum processing conditions of temperature and time for extraction. Optimization results were validated and verified; optimized processing conditions resulted in increasing the yield of extract up to 22% from the plant’s dried leaves without affecting the cytotoxic activity of the extract. The crude extract was further separated and purified using thin layer chromatography, chlorophyll removal techniques and column chromatography. In order to get the compound with highest anticancer activity, the fractions obtained from each step of separation and purification was tested for their antiproliferative activity against human breast cancer MCF-7 cell line and normal Vero cell line using In vitro MTT assay. The pure fraction obtained from column chromatography coded as PISAC2.3 exhibited an IC50 value of (2.8 µg/ml) and (88.6 µg/ml) against human breast cancer cell line and normal Vero cell line respectively, which indicates a high cytotoxicity against cancer cells and safety toward normal cells. The compound was identified using nuclear magnetic resonance (NMR); NMR results showed that the compound is an unsaturated fatty acid with a molecular formula of (C11H20O2). In conclusion, the identified compound requires further in vivo studies, as it is a potential possible anticancer drug due its high activity against cancer cells and also its safety for normal cells.