Abstract

This study was designed to search for novel anti-cancer compounds from natural plants. The 70% ethanolic extract from the rizhomes of Cimicifuga dahurica (Turcz.) Maxim. (Ranunculaceae) was found to possess significant in vitro anti-proliferative effects on MCF-7 breast cancer cells. A phytochemical investigation using assay-guided fractionation of the ethanolic extract of C. dahurica resulted in the isolation of one new phenolic amide glycoside 3, two new lignan glycosides 4 and 7, one new 9,19-cycloartane triterpenoid glycoside 6, and thirteen known constituents 1, 2, 5, and 8–17. The structures of 3, 4, 6, and 7 were established using contemporary NMR methods and from their HRESIMS data. The anti-proliferative effects of isolated compounds were evaluated using the BrdU-proliferation kit. Five among the 17 isolated compounds showed significant anti-proliferative effects (p ≤ 0.05), wherein compound 7 showed the most significant anti-proliferative and cell cycle arresting effect (p ≤ 0.05) which followed a dose dependent manner. Western blot protein expression analysis showed a down expression of c-Myc and cyclin D1 which further elucidated the anti-proliferation mechanism of compound 7 while apoptotic effects were found in association with Bcl-2 family protein expression variations. Conclusively this study reports the isolation and identification of 17 compounds from C. dahurica, including four novel molecules, in addition to the fact that compound 7 possesses significant anti-proliferative and apoptotic effects in vitro that may require further exploration.

Highlights

  • The genus Cimicifuga has been widely used all over the world and since ancient times has been a traditional Chinese herbal medicine

  • MCF-7 breast cancer cells were exposed to epidermal growth factor (EGF) which was known as a stimulator of proliferation

  • The cell cycle arrest at G0/G1 phase was reversed by a pan caspase inhibitor or a caspase 3 inhibitor but not the nucleoside addition. These results demonstrated that pan caspase and caspase 3 maybe had major roles in the mechanism of cell cycle arrest effects of compound 7 on MCF-7 cells

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Summary

Introduction

The genus Cimicifuga ( known as Actaea) has been widely used all over the world and since ancient times has been a traditional Chinese herbal medicine. This genus consists of 28 species worldwide [1], among which nine species are indigenous to China. In traditional Chinese medicine, Shengma is widely used for treating aphtha, sore throat, toothache, and wind-heat headache. It has been used in archoptosis, non-erupting measles, spot poison, uterine prolapse, and other diseases [1,2,3,4].

Characterization of Compounds 1–17
Cell Proliferation Activity
Cell Cycle Distribution and Western Blot Analyses
Molecules maybe had
General Experimental Proceduces
Plant Material
Extraction and Isolation
Acid hydrolysis and Sugar Identification
Cell Culture
Anti-Proliferative Activity
Cell Cycle Distribution Analyses
Western Blotting
Statistics

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