CHAPTER 1

INTRODUCTION

1.1 Research background

Cancer is one of the most serious, complex and diverse diagnoses to receive and

has been a leading cause of death worldwide. Because of its high mortality rate, plenty

of anticancer drugs have been employed in cancer treatment to inhibit the proliferation

of cancer cells (Wilkes & Ades 2004). Nevertheless, many of these agents would kill

normal cells and have side effects on patients who undergone the treatment for instance

hair loss, tiredness, vaginal discharge, muscle pain, hot flashes, burning, vomiting,

nausea and dizziness (Holly 2009). Thus, this research had been done to explore the

capability of selected Malaysian ‘ulams’ in inducing antiproliferative activity selectively

to cervical cancer cell lines, HeLa.

In 2003, a total of 21,464 cancer cases were diagnosed among Malaysians in

Peninsular Malaysia, comprising 9,400 males and 12,064 females. The commonest

cancer in males was lung cancer (13.8%). Whereas among females, the frequent cancers

were breast cancer (31.0%) and cervical cancer (12.9%) (Lim & Halimah 2004). There

were a total of 1,557 cases of cancer cervix in Malaysia (ASR of 19.7 per 100,000

population), consisting of Chinese women with highest ASR of 28.8 per 100,000

population, followed by Indians with ASR of 22.4 per 100,000 population and Malays

with ASR of 10.5 per 100,000 population (Lim & Halimah 2004).

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 For decades, efforts to find anticancer agent from natural sources like

microorganisms, marine and plants have been received much attention (Fingrut &

Flescher 2002; Yuan & Walsh 2006; Ren et al. 2008; Choi & Lee 2009). Many

researches have been carried out to hunt medicinal plant bioactive compounds with

potential in cancer treatment. Bioactive compounds such as alkaloid, flavonoid, volatile

oils, resin, saponin, steroid and tannin were consumed as a drug that contain

antiproliferative effects which induces apoptosis to cancer cells in cancer treatment (Ong

2004).

In Malaysia, the uses of ‘ulam’ and herbs have long been used traditionally for

the treatment of various diseases (Rukayah 2000; Ayob 2001; SMSI & Fauziah 2008).

‘Ulam’ is a plant that eaten raw or soak in hot water before being eaten. It is very

popular among Malaysians and usually eaten together with gravy such as ‘budu’,

‘cencaluk’ or ‘sambal belacan’. ‘Ulam’ came from many parts of plants including young

leaf, seed, fruit, stem, rhizom, tuber and flower (Rukayah 2000). Built from valuable

bioactive compounds, it contains high minerals and vitamins as calcium, phosphorus,

iron and fibre which are important for human digestive. These advantages have

encourage this research to screen for anticancer properties of four Malaysian ‘ulams’

namely, Syzygium polyanthum (serai kayu), Barringtonia racemosa (putat), Oroxylum

indicum (beko) and Vitex pinnata (leban) towards cervical cancer cell lines, HeLa.

Cancer can be defined as a group of diseases which is characterized by

uncontrolled cellular growth without respect to normal limits or suppressed apoptosis,

leading to excessive proliferation and spread of cells to other locations in the body

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(Nevidjon & Sowers 2000). Antiproliferative characteristic is needed for drug in cancer

treatment because this could inhibit the uncontrolled proliferation of cancer cells and

create the antiproliferative with apoptosis. Apoptosis is a form of programmed cell death

caused by activation of endogenous molecules (e.g. p53, Bax, caspase) leading to the

fragmentation of DNA (Lockshin & Zakeri 2004).

The objective of cancer treatment is to induce cancer cells death selectively

without affecting normal cells. Several therapies have been done in combating cancer

such as chemotherapy which has been used in treatment of choice for malignant

lymphoma and leukemia (Parakama & Clive 2001). However, the side effect has pushed

many researchers to find a new drug from plants derivatives that can treat complications

arise from cancer therapy. A good antiproliferative drug should inhibit almost all of the

cancer cells. This could be determined through the IC50 value of the plant extract. IC50 is

the concentration that needs to inhibit 50% of cancer cells proliferation from whole cells

population. In this study, the four Malaysian ‘ulams’ will be screen for its anticancer

potential through antiproliferative assay and apoptosis mechanisms study.

1.2 Objectives of the research

1) To determine IC50 value of ‘ulam’ extracts that have selective antiproliferative and

cytotoxycity effects on cervical cancer and normal cell lines.

2) To characterize the death mechanisms of ‘ulam’-treated cells.

3) To study the pro-apoptotic and anti-apoptotic proteins in plant-treated cells.

4) To detect the different classes of secondary metabolites in active ‘ulam’ extract.

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1.3 Significance of the research

This research was enabled thorough screening, identification and evaluation of

the Malaysian ‘ulams’ and their bioactive compounds known or having the potential in

cancer treatment. In addition, this research provided preliminary data and also the

establishment of cancer cell lines for further research which will lead to new

understanding in cancer research and development of drugs from medicinal plants. The

project is anticipated to make substantive contribution to the field of anticancer research

in Malaysia by identifying the new plants and their bioactive compounds with antitumor

and anticancer properties. The direct beneficiary of the project will be the patients who

are suffering from cancer and also the government in saving currency exchange due to

the import of anticancer drugs. The data obtained will also benefit MIGHT, MARDI,

scientific community, traditional medicine practitioners in developing new anticancer

drugs and their administration.

1.4 Hypothesis of the research

1) ‘Ulam’ extract have antiproliferative activity selectively on cervical cancer cell lines

without having cytotoxycity effect on normal cell lines and IC50 value is determined.

2) ‘Ulam’-treated cells are dead through an apoptosis pathway.

3) ‘Ulam’-treated cells express the involvement of pro-apoptotic (e.g. Bax, p53,

caspase) and anti-apoptotic proteins (e.g. Bcl-2, E6) in apoptosis process.

4) Several classes of secondary metabolites (e.g. alkaloid, flavonoid, resin, saponin,

steroid, tannin) in active ‘ulam’ extract are detected.