Anticancer Activities of Extracts and Purified Compounds from Jeruju Putih (Acanthus Ilicifolius L.)
Currently, cancer is Malaysia’s number two killer only preceded by cardiac disease. Unfortunately, there is no effective cure for the disease has yet to be discovered. The diverse local native plants of Malaysia offers an avenue in developing herbal medicines especially for treating chronic dise...
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| Format: | Thesis |
| Language: | English English |
| Published: |
2007
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| Online Access: | http://psasir.upm.edu.my/id/eprint/7194/1/FPSK%28P%29_2007_4a.pdf http://psasir.upm.edu.my/id/eprint/7194/ |
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| Summary: | Currently, cancer is Malaysia’s number two killer only preceded by
cardiac disease. Unfortunately, there is no effective cure for the disease
has yet to be discovered. The diverse local native plants of Malaysia
offers an avenue in developing herbal medicines especially for treating
chronic diseases such as cancer. Acanthus illicifolius L. is a mangrove
plant, which can be commonly found thriving in Malaysian coastal
swamps. The plant is commonly (known as Holly-Leaved mangrove and
locally) known as jemuju, jeruju or daruju. It has been used traditionally
for generations as a treatment for various diseases. The objectives of the
study are to separate pure compounds from Jemuju seed and to
determine anti-proliferation activity and mechanism pathway of the action
pure compounds. Four types of Jemuju seed extracts were separately
prepared using distilled water (SA) and organic solvents, which include
successively with hexane (FH), ethyl acetate (FE) and methanol (FM).
Anti-proliferation activity tests of these extracts were carried out on seven types of cancer cell lines (HeLa, CEM-SS, CaCO-2, HEp-G-2,
CaOV-3, MDA-MB-231 and MCF-7). The SA extract was the most active
on the HeLa cells (EC50 101 ± 15 μg/ml) while less activity was found on
the other test cells. FE extract was most active on CEM-SS cells with an
EC50 value of 39.33 ± 4.16 μg/ml followed by HEp-G-2, CaCO-2, CaOV-
3, MCF-7, MDA-MB-231 and HeLa cells, and not activities on MCF-10A
cells. On the other hand, no activities were observed for FH and FM
extracts. None of the extracs were active on the MCF-10A normal human
breast cell line. Results on apoptosis test using DNA ladder and AO/PI
methods demonstrated that FE caused internucleosomal DNA damage
by causing DNA fragmentation on the cells treated with 40, 60, 80 or 100
μg/ml of this extract compared with non-treated control cells. Using the
AO/PI method, a higher percentage of cells which underwent apoptosis
were observed in the cells treated with 80 or 100 μg/ml, with EC50 values
of 73.91 ± 1.55 and 97.63 ± 1.46%, respectively. On the other hand, only
1.92 ± 0.17% cells underwent apoptosis in non-treated cells. The
purification process of FE using liquid chromatography produced 13
fractions where four of the fractions (FE-3, FE-5, FE-6, FE-10) showed
high anti-proliferation activity towards MCF-7, MDA-MB-231, HeLa and
CaOV-3 cells. Three of the fractions (FE-3, FE-6, FE-10) were further
purified using column chromatography, to yeild a total of 67 fractions of
which 32 were found to be active against cancer cell lines. The process
of re-crystalisation was carried out on the FE-5 fraction until white
crystals were produced. Four compounds were successfully purified,
their molecular structures have been determined and labeled as J1 (Benzozaxolyne-2-on), N4 (N-benzoxazolol), NW5 (4-hydroxy-3Hbenzoxal-
2-on) dan NW7 (2-hydroxy-2H-1,4-benzoxazin-3(4H)-on). Due
to insufficient amount of compounds the molecular structures for the
other five compounds could not be obtained. Anti-proliferation activity
tests of the pure compounds on five types of cell lines (MCF-7, MDA-MB-
231, HeLa, CaOV-3 and MCF-10A) showed that J1, NW2 and NW3,
have no effects. Compounds N3, NW5 and NW7 showed high
antiproliferation on activity towards MCF-7, with EC50 values of 7.50 ±
0.32 μg/ml, 8.50 ± 0.21 μg/ml and 7.50 ± 0.65 μg/ml, respectively. NW14
also showed high antiproliferation activity towards MCF-7 and HeLa with
EC50 values of 8.00 ± 0.35 μg/ml and 8.21 ± 1.08 μg/ml respectively.
Importantly, all of the pure compounds obtained clearly indicated no
activities on MCF-10A, which is normal human breast cell line. EC50
values for tamoxifen which was used as positive controls on MCF-7,
MDA-MB-231, HeLa and CaOV-3 cells were also obtained. Suppression
of oncogene expression by three of the active compounds namely NW5,
NW7 dan N4 was also monitored on selected cells. NW5 was found to
suppress oncogene expression in MCF-7, MDA-MB-231, HeLa dan
CaOV-3. NW7 suppressed oncogene expression in MCF-7 and HeLa
cells, while N4 was able to suppress oncogene expression in MCF-7.
Thus all compounds tested exhibited suppression of oncogene c-erb-B-2
expression as the mechanism of action on all treated cells. Since the
potential of jemuju has been demonstrated, it is hoped that more indepth
studies on the plant will continue to be pursued. |
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