Chemical constituents of endophyte Myrothecium roridum LLY from Ephedra equisetina and their in vitro cytotoxicity
10.7501/j.issn.0253-2670.2015.21.005
- Author:
Li SHEN
1
Author Information
1. Medical College of Yangzhou University
- Publication Type:Journal Article
- Keywords:
Cytotoxicity;
Deoxyleporin B;
Endophyte;
Ephedra equisetina Bge.;
N-methyl-1H-indole-2-carboxamaide
- From:
Chinese Traditional and Herbal Drugs
2015;46(21):3155-3161
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To study the chemical constituents of solid fementation product of endophyte Myrothecium roridum LLY from Ephedra equisetina and their in vitro cytotoxic activity against cancer cells. Methods: Isolation and purification were performed through silica gel and Sephadex LH-20 gel filtration column chromatography as well as HPLC method, and structures of obtained compounds were elucidated by HR-MS and NMR comparised with the data of literatures. In vitro cytotoxicity was evaluated through MTT method and preliminary cytotoxicity mechanism was studied using flow cytometry and Western blotting. Results: Nine compounds were isolated from the solid fementation product of endophyte M. roridum, which were identified as 1-phenazinecarboxylic acid (1), 1-methoxyphenazine (2), N-methyl-1H-indole-2-carboxamaide (3), monomethylsulochrin (4), ditryptophenaline (5), lumichrome (6), deoxyleporin B (7), uracil (8), and thymine (9), respectively. In vitro cytotoxicity assay showed that compound 3 was cytotoxic to the human hepatoma cell line SMMC-7721 with an IC50 value of 38.0 μg/mL, while positive control cisplatin had an IC50 value of 11.5 μg/mL. Cell cycle analysis indicated that compound 3 could delay SMMC-7721 cells in S phase and Western blotting showed that compound 3 could up-regulate the expression of cell cycle negative regulation protein p27 and induce the cleavage of PARP. Conclusion: Compound 7 is a new natural product. Compound 3 could induce the apoptosis of SMMC-7721 cells and the anti-proliferation activity of compound 3 to SMMC-7721 cells is related to S phase arrested and regulated by protein p27.
