Anti-proliferative and apoptotic activities of constituents of chloroform extract of Juglans regia leaves.
OBJECTIVE
To evaluate anti-proliferative as well as apoptotic activities of compounds identified in chloroform extract of Juglans regia leaves, on human breast and oral cancer cell lines (MCF-7 and BHY).


METHODS
Column chromatography, MTT assay, flowcytometry and western blotting have all been used in the study.


RESULTS
Bioassay-guided fractionation of chloroform extract of J. regia afforded isolation of 5-hydroxy-3,7,4'-trimethoxyflavone [1], lupeol [2], daucosterol [3], 4-hydroxy-α-tetralone [4], β-sitosterol [5], 5,7- dihydroxy-3,4'-dimethoxyflavone [6] and regiolone [7]. Structures of the compounds were established on the basis of spectroscopic analyses [Nuclear magnetic resonance (NMR) and mass]. All compounds inhibited proliferation of MCF-7 (human breast adenocarcinoma) and BHY (human oral squamous carcinoma) cells in a concentration-dependent manner. Compounds 6 and 7 had potent cytotoxic effects on both MCF-7 and BHY cells (IC50 21-51 μm), yet were not toxic to normal cells. MCF-7 growth inhibition was attributed to apoptosis; population of apoptotic cells increased from 1.12% in controls to 5.64 and 8.1% after 48-h treatment with compounds 6 and 7, indicating their potential at inducing early and late apoptosis. The caspase cascade was not activated, as indicated by only insignificant cleavage of caspase-3.


CONCLUSIONS
Our results suggest that compounds 6 and 7 can induce apoptosis in MCF-7 cells through the caspase-3 independent pathway.
Department of Physiology and Pharmacology, Pasteur Institute of Iran, Tehran, 3159915111, Iran.
M
M
Salimi
M H
MH
Ardestaniyan
H
H
Mostafapour Kandelous
S
S
Saeidnia
A R
AR
Gohari
A
A
Amanzadeh
H
H
Sanati
Z
Z
Sepahdar
S
S
Ghorbani
M
M
Salimi
D000230:Adenocarcinoma / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D002725:Chloroform; D005260:Female; D047309:Flavones / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D031324:Juglans / Q000737:chemistry; D061986:MCF-7 Cells; D009062:Mouth Neoplasms / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D008517:Phytotherapy; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology; D018515:Plant Leaves / Q000737:chemistry; D015398:Signal Transduction / Q000187:drug effects
D000972:Antineoplastic Agents, Phytogenic; D047309:Flavones; D010936:Plant Extracts; D002725:Chloroform; C505407:CASP3 protein, human; D053148:Caspase 3