28-29
2-3
31
5
33-34
7
9-15
9-15
25-26
INTRODUCTION
• Fragrant rice MRQ76 is specialty rice declared by the Malaysian
Agricultural Research and Development Institute (MARDI) in 2012 and it
has similar characteristics to Jasmine fragrant rice, and Khao Dawk
Mali 105 imported from Thailand.
• In this study, the unpolished and polished grains of the fragrant rice
MRQ76 that aged 3 months were used to look at the anti-proliferative
activity on human adenocarcinoma cancer cell, HT-29.
• Unpolished rice grains provide several bioactive compounds such as
phenolic acids, flavonoids, γ -oryzanol, and Gamma aminobutyric acid
(GABA) and it also has high fiber contents (Malabadi, n.d.).
• Polished rice is a white rice produced after the outer layers of brown rice
are removed during milling process (Rathna Priya et al., 2019).
• In 2020, a study by Rohin et al. conclude that organic rice MRQ74 has
anti-proliferative activity against colon cancer cells by the formation of
apoptotic bodies and reduction in cells viability through AO/PI staining.
2
General Objective
To examine the anti-proliferative activity of unpolished and
polished sample of fragrant rice MRQ76 that aged 3 months on HT29 colon cancer cell line.
Specific Objective 1
To evaluate the anti-proliferative activity of unpolished and polished
fragrant rice MRQ 76 on human adenocarcinoma colon cancer cell line.
Specific Objective 2
To observe morphological changes of human adenocarcinoma colon
cancer cell line after being treated using fragrant rice water extract and its
association with the anti-proliferative activity.
Specific Objective 3
To determine the correlation between IC50 values and anti-proliferative
activity of fragrant rice MRQ 76 on human adenocarcinoma cancer cell
line, HT-29.
Hypothesis
Null hypothesis
There is no significant difference in the
anti-proliferative activity of fragrant rice
MRQ 76 on colon cancer cell, HT-29.
Alternate Hypothesis
There is a significant difference in the
anti-proliferative activity of fragrant rice
MRQ 76 on colon cancer cell, HT-29.
3
LITERATURE
REVIEW
Author
Rohin et al.
Abubakar
al.
Malabadi
et
Importance of the
study/review
Study objective
The consumption of organic
rice
MRQ74
showed
preliminary results on cancer
study.
To examine the anti-proliferative
properties off different extracts of
new variety an organic rice MRQ74
towards colon cancer cells
The germinated brown and
brown forms of fragrant rice
have higher DPPH radical
scavenging ability than white
form fragrant rice
To investigate the composition of
several
white,
brown
and
germinated brown rice cultivars in a
bid to offer insight into the growing
diet-related metabolic diseases in
regions where rice is a staple food
Brown or unpolished rice have
higher levels of nutrients such
as protein, fat, minerals, and
vitamins than those in polished
or milled white rice.
To highlights the dietary importance,
arsenic toxicity, and nutritional value
of brown, white, basmati and
pigmented rice
Method
Result
double
The new variety of organic rice
MRQ74 extracts possesses antiproliferative activity against HCT116, CT-26 and HT-29
• DPPH
radical
scavenging ability
• TPC
• TFC
• Proximate analysis
The properties of germination
and high amylose content, if
present in a rice cultivar would
less predispose persons who
depend on them as a staple food
to
diet-related
metabolic
diseases.
• MTT assay
• AO/PI
staining
5
METHODOLOGY
(Rohin et al., 2018)
(Mosmann, 1983)
(Melini et al., 2017)
(Rohin et al., 2020)
(Merlin et al., 2010)
(Hajiaghaalipour et al., 2017)
7
STATISTICAL
ANALYSIS &
RESULTS
9
DISCUSSIO
N
• Table 1 shows the yield of polished and unpolished MRQ76 aging rice
samples extracted in deionized water.
• Water extraction was used because it is cheap, non-toxic and nonflammable.
• Unpolished sample gave higher extraction yield, i.e., 1.98% compared to
polished sample with 1.12%.
• Independent T-test result show that the extraction yield is statistically
significant different from each other.
10
11
DISCUSSIO
N
• Table 2 shows the IC
72 hours.
50
values (mg/mL) of HT-29 cell line by MRQ76 rice extracts at
• The study evaluate the anti-proliferative activity of unpolished and polished
fragrant rice MRQ 76 (1.00 - 0.016 mg/mL) was investigated using MTT assay.
• According to the National Cancer Institute standards, crude extracts’ possessing
an IC50 value of less than 20 μg/mL only considered to be active potential
cytotoxic extract against tested cancer cells (Geran et al., 1972)
• Therefore, the extracts possess anti-proliferative properties by inhibiting the
proliferation of cell line at IC50.
• All extracts only show anti-proliferative activity at 72 hours after treatment with
unpolished sample possess lower IC50 values than polish sample with 47% and
41% respectively (p<0.05).
• However, the drug sample, 5-FU lowest IC50 values, i.e., 33%.
12
• 5-FU is a commercial drug commonly used to treat cancer and it is more effective
in inhibiting the proliferation of treated cancer cells compared to the rice
extracts.
• A study by Rohin et al., 2020 showed that organic rice MRQ74 has antiproliferative properties toward colon cancer cells attributed by its polyphenols
content including phenolic acid such as p-coumaric acid, ferulic acid, vanilic acid
and caffeic acid (Rohin et al., 2020)
• Unpolished sample have more nutritional values than polished sample, supposed
to explained the result.
13
14
DISCUSSIO
N
• Table 3 shows the correlation between IC
values and samples’ concentrations to
anti-proliferative activity by MRQ76 rice extracts at 72 hours
50
• All the negative correlation means that as the concentration increased, the IC50
values decreased, decreased IC50 indicated that there is anti-proliferative activity
of this rice.
15
16
(i)
(ii)
(iii)
(iv)
Cells densed
and lobulated
Cells shrinkage or
reduced cell sizes
Healthy and intact
Apoptosis bodies
17
(i)
(iii)
(ii)
Healthy and intact
Cells shrinkage or
reduced cell sizes
(iv)
Cells densed and lobulated
Apoptosis bodies
18
(i)
Healthy and intact
(iii)
(ii)
(iv)
Cells shrinkage or
reduced cell sizes
Cells densed and lobulated
Apoptosis bodies
19
(a
)
(i)
(ii)
(iii)
Viable cells
Viable cells
Viable cells
(b)
Death cells
Early apoptosis
Late apoptosis
Late apoptosis
Early apoptosis
Late apoptosis
20
(c)
Late apoptosis
Late apoptosis
Late apoptosis
Death cells
Early apoptosis
Death cells
(d)
Late apoptosis
Late apoptosis
Early apoptosis
Death cells
Early apoptosis
Late apoptosis
21
DISCUSSIO
N
• Cell proliferation also can be observed by apoptosis or cell death that analyzed
using morphology observation and AO/PI staining procedures.
• Apoptosis can be defined as a defense mechanism against cancer that destroys
own cell and it have several stages starting from cell shrinkage, cells
fragmentation, cytoskeleton collapses, nuclear envelope disassembles and lastly,
cells release apoptotic bodies (Apoptosis,2022).
• Figure 1,2 and 3 show morphological changes of untreated HT-29 cell and HT-29
cell after being treated with MRQ76 rice extracts and 5-FU at 24 hours, 48 hours
and 72 hours.
• The morphological changes are associated with the IC50 values.
• Figure 1,2 and 3 (i) are untreated cells and it show healthy and intact morphology
with 90% confluence and the cell still detach to the wall.
• The cell are viable as can be seen through A0/PI staining with present of all green
fluorescence color shown in Figure 4 (a)(i)(ii)(ii).
22
• Figure 1,2 and 3 (ii) are treated cells after 24 hours treatment, and the morphology
show cell shrinkage and reduction of the cell sizes for all treated cells.
• The cells treated with 5-FU enter early stage of apoptosis and some already gone
through late apoptosis after 24 hours as there are green and orange fluorescence
color appeared during the staining (Figure 4 (b)(i)).
• For 5-FU that has the lowest IC50 values, apoptosis bodies appeared which is the
last stage of apoptosis with the appearance of debris in the background (Figure 3
(iv) and there are a lot of death cells as shown by the red fluorescence color in
Figure 4 (b)(iii)
• Unpolished rice (Figure 4 (d)(iii)) show more dead cells than polished rice (Figure 4
(c)(iii)) as it has higher IC50 values as shown in Table 2.
23
CONCLUSION
• This is the first study to examine the anti-proliferative activity of unpolished
and polished sample of fragrant rice MRQ76 on HT-29 colon cancer cell line.
• The results exhibited that unpolished rice has higher anti-proliferative activity
on the cancer cell than polished rice.
• The IC50 values and the morphological changes represents the antiproliferative activity of each sample.
• Thus, the objectives of this study are achieved.
• To evaluate the anti-proliferative activity of unpolished and polished fragrant
rice MRQ 76 on human adenocarcinoma colon cancer cell line (Table 2).
25
• To observe morphological changes of human adenocarcinoma colon cancer cell
line after being treated using fragrant rice water extract and its association with
the anti-proliferative activity (Figure 3).
• To determine the correlation between IC50 values and anti-proliferative activity
of fragrant rice MRQ 76 on human adenocarcinoma cancer cell line, HT-29 (Table
3).
• This study shown that fragrant rice MRQ76 has potential anti-proliferative activity
on HT-29 colon cancer cell line.
• In conclusion, this rice variety has potential as cancer treatment, but further
studies should have been done before it officially declared in the industry.
26
LIMITATION,
STRENGTHS AND
RECOMMENDATION
S
LIMITATIONS
• There are still lack of research regarding fragrant rice MRQ76 - including antiproliferative activity.
• The study involve direct interaction with cancer cells that are very sensitive need to work in sterile environment to avoid contamination.
• Time consuming
• Equipments - some analysis are done manually
STRENGTH
Sof unpolished and polished sample - have different nutrient values
• The use
• Comparison with 5-FU drug
28
• Study on specific bioactive compounds in fragrant rice MRQ76
• Explain the anti-proliferative effects in this study.
• Study on cytotoxic activity of fragrant rice MRQ76 with several solvent
extraction
• Further analysis of fragrant rice MRQ76 on cancer treatment study will
increase its potential as alternative treatment for cancer.
• Study on how harvest periods effects the anti-proliferative activity of
fragrant rice MRQ76
• Aged rice samples might have different anti-proliferative activity
compared to fresh rice samples.
29
• Kelantan Agricultural Group Berhad
• Indonesia-Malaysia-Thailand Growth Triangle (IMT-GT)
• Malaysia Agricultural Research and Development Institute (MARDI)
31
REFERENCES
Abubakar, B., Yakasai, H. M., Zawawi, N., & Ismail, M. (2018). Compositional analyses of white, brown and germinated forms of
popular Malaysian rice to offer insight into the growing diet-related diseases. Journal of Food and Drug Analysis, 26(2), 706–715.
https://doi.org/10.1016/j.jfda.2017.06.010
Apoptosis.
(2022).
Retrieved
21
May
2022,
from
https://www.genome.gov/geneticsglossary/apoptosis#:~:text=Apoptosis%20is%20the%20process%20of,have%20been%20damaged%20beyond%20repair.
Geran, R. I., Greenberg, H. M., McDonald, M., (1972). Protocols for screening chemical agents and natural products against animal
tumours and other biological systems. Cancer Chemoth
Hajiaghaalipour, F., Faraj, F. L., Bagheri, E., (2017). Synthesis and characterization of a new benzoindole derivative with apoptotic
activity against colon cancer cells. CurrPharm Des.
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morphological study of different Duku(Lansium domesticum Corr.) extract toward human colorectal adenocarcinoma cells line
(HT-29). Pharmacognosy Journal.
Malabadi, R. B. (n.d.). White, and Brown Rice-Nutritional value and Health benefits; Arsenic Toxicity in Rice plants Synthesis of
metal oxide nanoparticles using green route View project. https://www.researchgate.net/publication/362516290
Melini, V., Acquistucci, R., (2017). Health-promoting compounds in pigmented Thai and wild rice. Foods.
Merlin, N. J., Parthasarathy, V., Santhoshkumar, T. R., (2010). Induction of apoptosis in human breast cancer cell line MCF-7 by
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Rathna Priya, T. S., Eliazer Nelson, A. R. L., Ravichandran, K., & Antony, U. (2019). Nutritional and functional properties of coloured
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Rohin, M., Hadi, N., & Ridzwan, N. (2020). Anti-proliferative Properties of New Variety Organic Rice MRQ74 Extracts against Colon
Cancer Cells: In-vitro Study
34