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Characterızatıon Of Musk Wıllow Extract For Antıcarcınogenıc Phytochemıcal Compounds

During past decades, several naturally occurring compounds have been studied in terms of their potential chemopreventive role in colorectal cancer development and various in vitro and in vivo models supported their antiproliferative and cancer preventive abilities. One such candidate is an extract from willow species. Musk Willow, a Salix species endogenous to the Middle East, has not been examined to date for its anticarcinogenic effects. We have previously shown that the extracts from these plants have high amounts of salicin, potent antioxidant activity and significant amounts of phenolic compounds famed for their immunomodulatory and anti-inflammatory activities (Enayat and Banerjee 2009). Sustaining proliferative signaling, evading growth suppressors, utilizing mechanisms to escape from programmed cell death (apoptosis), enhanced motility and ability of invasion as well as anchorage independent growth are some of the major hallmarks of carcinogenesis. Therefore, in order to study the anticarcinogenic effect of extracts from Musk Willow we treated HCT-116 and HT-29 colon cancer cell lines as well as a non-transformed colon fibroblast CCD-18Co cell line with the ethanolic extract of bark. Our preliminary data indicate that the extract exerted strong antiproliferative, proapoptotic and antimetastatic effects on both colon cancer cell lines without being toxic for nontransformed colon fibroblast cells. Therefore, we are proposing to further investigate the anticarcinogenic role of extract from Musk Willow and analyze the crude extract for its active constituents which could be responsible for the observed antitumorogenic effects in HCT-116 and HT-29 colon cancer cell lines. Towards this objective, we propose to: • Confirm the effect of the extract on apoptosis by Annexin V apoptotic assay using Flow cytometry • Determine the effects of the extract on the cell cycle by the cell cycle analysis using Flow cytometry. • Determine the antimetastatic potential of the extract by examining its effect on motility and invasion of cancer cells using an in vitro wound healing assay and Invasion through Matrigel respectively. • Determine the effects of the extract on anchorage independent growth of the extract by soft agar colony formation assay. • Partition the ethanolic extract of bark of Willow into petroleum ether, ethyl acetate, butanol and water fractions by solvent-solvent fractionation • Define the bioactivity of each fraction on HCT-116 and HT-29 colon cancer cell lines using MTT cell proliferation assay.