Introduction1, INTRODUCTION Background Cancer is a major killing disease in most of the developed and developing countries. It is a growing public menace. Ten million new cases of cancer and 6.4million deaths due to cancer were reported by WHO, in 1997. Particularly breast cancer is the most common malignancy affecting female population and the third most common cancer in the world with about 48,000 women die because of breast cancer every year. Over 50 % of breast cancer incidence occurs in the developed countries (Castro et al., 2001). In the past few years, incidence of breast cancer has been dramatically increasing in developing countries (Anisimov, 2003). During the last two decades the percentage of occurrence of cancer has been high for breast cancer in female population. Definition It is a well known that Cancer is a progressively debilitating disease, which is characterized by a persistent, abnormal, and relatively autonomous proliferation of cells. This may be the result of a permanent cellular defect that is passed on to the cell progeny. The defect may be due to a single or a combination of several factors and once developed, usually becomes independent on them (Diamandopoulos and Meissmes, 1985).Carcinogenesis is recognized as a multistep process, which involves mutations, failure of DNA repair mechanism and activation of oncogenes and loss of tumor suppressor function. The appearance of the disease in patients is the last stage of this process (Russo ef al., 1982). Breast cancer begins as a small painless lump. Whereas, metastatic breast cancer is currently an incurable, yet treatable disease, while Median survival is 18-24 months. Survival ranges from a few weeks to several years. The metastatic breast cancer remains as a chronic relapsing and remitting disease. It may respond for a time to an assay of cytotoxic anticancer drugs. Classification of Cancer Tumors are classified based on their invasiveness into two types ie. benign and malignant tumors. Benign tumors Benign tumors do not penetrate (invade) into adjacent tissue borders, nor do they spread (metastasize) to distant sites. These tumors are more differentiated and they closely resemble their tissue of origin. Malignant tumors Malignant tumors have the added property of invading contiguous tissues and metastasing to distant sites, where subpopulations of malignant cells take up residence, grow anew and again invade (Rubin and Farber, 1990). As a general rule malignant tumors kill and benign ones do not.Historical Background Tumor may occur at all times in human and animals in all parts of the world. The earliest evidence available at present is found in the form of tumor in human bones from pre-historic and historic periods. Reference is made in it to tumor treatment, either with knife or with an arsenical compound ic. chemotherapy. Descriptions relating to malignant tumor are also found in various Egyptian papyri (1500 BC). The Egyptians were the first to realize that tumor arising in various parts of the body differed in their behavior and should be treated differently. Hippocrates of Cos (460-375 BC), the father of medicine, coined the term “Karkinoma” for solid malignant tumors. The term “cancer” appeared much later, derived from the Latin word “Cancrum” denoting “crab” (Diamanopoulos and Meissner, 1985). It was not until greater understanding of the circulatory system was added to the body of medical knowledge in the 17" century that the early physicians made the link to the lymph glands in the armpit. The French surgeon Jean Louis Petit (1674-1750) and later the Scottish surgeon Benjamin Bell (1749-1806) were the first to remove the lymph glands, breast tissue, and underlying chest muscle. William Stewart Halsted started performing mastectomies in 1882 carried on their successful work. He became known for his Halsted radical mastectomy, a surgical procedure that remained popular up to the 1970's.Causative Factors of Cancer The causative factors of most cancers are not identified and the mechanism of carcinogenesis remains unclear. Clinical and scientific observations over the centuries have revealed many facts like the influence of chemical, viral, physical and genetic factors on carcinogenesis. A number of food additives, pesticides and industrial chemicals introduced commercially in the last fourty years have proven to be carcinogenic both in animals and in human systems (IARC monographs, 1971-1985). Types of Cancer Cancer is classified into 4 major groups as, 1. Carcinoma 2. Sarcoma 3. Lymphoma 4, Leukemia Carcinoma: Most of the human’s cancers are carcinoma. Carcinoma develops from epithelial cells of ectodermal or endodermal origin. For e.g. carcinoma of the brain, skin and breast can be stated. Sarcoma: Only 2% of cancer falls on sarcoma grouping. It is a malignant neoplasm of the soft connective tissue arising from fibrous, muscular, synovial, vascular or neural tissues, which are mesodermal in origin.Lymphoma: Cancer of the lymphocyte producing organs such as lymph nodes and spleen are designated as lymphoma. 5% human cancers constitute lymphoma. Leukemia: Neoplastic growth of the leucocytes is called leukemia and 4% of the human cancer comes under this group. Epidemiology of Breast Cancer Study of malignant forms of cancer that develops in breast tissues causes mortality incidence, high risk factors and low neoplastic disease occurring in female population in the world. The study of clinical outcome in breast cancer such as symptomology and secondary prevention strategies are internationally well accepted findings from the epidemiological investigation. The experimental evidence for the chemotherapy and cancer prevention reveals the role of tobacco smoking, diet and environmental exposure. Breast is the most common site of cancer in women and its incidence is rising. Breast cancer rates have been climbing steadily in the United states and other industrialized countries since the 1940s, amounting to more than one million cases per year worldwide (Parkin et al., 2005). In United States, a women’s lifetime risk of breast cancer has nearly tripled during the past four decades. In 2005, an estimated 211,240, U.S women were diagnosed with invasive breast cancer. More than 58,000 women were expected to be diagnosed with one type of insitu breast cancer, meaning the tumor is confined to its original location in the breast. In 2005, breast cancer was expected to kill more than40,000 American women (American Cancer Society Cancer facts and figures 2005) and more than 410,000 women worldwide (Parkin et al., 2005). Breast cancer incidence in Europe is 1,80,000 cases per year and 20% of all malignancies in European women. One in 40 Japanese women developed breast cancer in their life span. Breast cancer is an exception before the age of 20 years, and below 30 years in rare cases but then, the incidence rises rapidly up to the age of 50 years after which the rate slows down, (although incidence rate continues to rise). Mortality rates of breast cancer in Wester Europe and North America are of the order of 15-25 per 1, 00,000 in women, which is 30- 40% of the incidence rate, while in the countries like India (23.5 %), Japan (33.4%), China (26.5 %) and Africa (29.5 %). In India, the incidence rates of breast cancer among 1 lakh female population were recorded as 25.6 %, 15.8 % and 20 % for Mumbai, Banglore, and Chennai respectively in 1985 (Alon, 1985). Breast cancer incidence constituted between 9.9 % (Dibrugarh) and 21.0 % (Mumbai) of female cancers in different hospitals based cancer registries in India. The International Agency for Research on Cancer (IARC), World Health Organization (WHO), International Association of Cancer Registries (IACR) and National Cancer Registries (NCR) in India are providing the periodical observation of worldwide cancer incidence from the hospital-based registries. These compilations of data represent the best sources of data for descriptive epidemiological studies of cancer.Causative Factors of Breast Cancer The Causative factors in breast cancer are complex and still not clearly understood. Over years, a number of factors have been identified which may play a important role in the etiology of breast cancer or associated with an increased risk of breast cancer development. The list of risk factors includes endogenous and exogenous factors, which are inseparable. Today, breast cancer, like other forms of cancer, is considered to be a tesult of damage to DNA. How this mechanism occurs comes from several known or hypothesized factors such as exposure to ionizing radiation. Some factors lead to an increased rate of mutation (exposure to estrogens) and decreased DNA repair which is influenced by the BRCA1, BRCA2 and p53 genes. Although many epidemiological risk factors, and biological co-factors and promoters have been identified, the majority of breast cancer incidence remains unattributable, and the primary cause is unknown. A significant environmental effect was revealed by the large difference in breast cancer incidence between countries and continents, and a migration effect which slowly increases the risk of breast cancer even across generations after migration from a country of lower incidence to a country of higher incidence, such as moving from China or Japan to the United States. Humans are not the only mammal prone to breast cancer. Some strains of mice, namely the house mouse (Mus domesticus) are prone to breast cancer which is caused by infection with the mouse mammary tumor virus (MMTV or "Bitmer virus" named after its discoverer Hans Bitter), by randominsertional mutagenesis. Suspicion of MMTV or other viruses in human breast cancer is controversial and the idea is not generally accepted for lack of direct and definitive evidence. Age The incidence of breast cancer continues to increase with age. A marked elevation in breast cancer incidence is observed in women in the age ranging from 45-55 years. In this context Pike ef al., (1993) are of the opinion that the incidence of the breast cancer around age group of 45-55 years may be due to the reproductive hormones, since non-hormone dependent cancer do not show these changes in incidence rate around the time of Menopause Breast Cancer incidence increases about 2.17% per year of age in the post-menopausal period. This rise can be attributed solely to endogenous estrogen (Pike et al., 1983, Shimizu et al., 1990). Age at Menarche The younger a woman’s age at menarche, the higher the risk of breast cancer (Klale G and Heuch I, 1988, Lavachia et al., 1985). The menstrual factors have an important role in the genesis of breast cancer. A number of workers (Mac Mohan et al.,, 1982, Aprer et al., 1989) have established that women with early menarche have higher estrogen level for several years after menarche and probably through-out their reproductive lives. In this connection Hsieb et al., (1990) found that for each 2 years delay in the onset of menstruation breast cancer risk was reduced by 10 %. Brinton et al., (1988) also found that women with onset of menstruation at or after the age of 15years had a 23 % lower risk than those with an age at menarche of 12 years or younger. Age at Menopause The later a woman’s age at menopause, the higher, the risk of breast cancer (Pike ef al., 1981, Trichopoulos et al., 1972). The increased risk is associated with early age of menarche and late age menopause suggested that the longer the exposure to sex hormones during the reproductive years, the higher the risk of breast cancer (Henderson et al., 1985). Age at Full Term Pregnancy The vast majority of studies have found that, the younger women who have full term pregnancy have low risk for breast cancer (Mac Mohan et al., 1970, Ewertz et al., 1990). Women who give birth to their first child after the age of 30 have a higher risk than nulliparous women (Layde et al., 1989). The average age at the full term pregnancy, another known risk factor has arisen dramatically in the last two decades Parity Nulliparous women are at increased risk for breast cancer in comparison with parous women (Pathak et al., 1986, Leon DA, 1989). The protective effect of multiparity has been noted mostly in women aged between 40-50 years or more.‘Geographical variation Studies of migrants from Japan and Hawaii shows that, the rate of breast cancer in migrants assumes the rates of the host country within one or two generation, indicating that the environmental factors are of great importance (Ries, 2005). Family history The incidence of 10 % of breast cancer in western countries is due to genetic predisposition. Breast cancer susceptibility is generally inherited as an autosomal dominant trait with limited penetrance. Two breast cancer susceptibility genes, BRCAl and BRCA2, account for a substantial proportion of very high risk families. Women with severe atypical epithelial hyperplasia have a 4-5 times higher risk of developing breast cancer. Women with palpable cysts, complex fibro adenomas, duct papillomas, etc. have a slightly higher risk of breast cancer (Loman et al., 2001: Lichtenstein et al., 2000). LIFE STYLE Diet The types of fat are important, although there is a close relationship between the incidence of breast cancer and dietary fat intake in populations, the true relationship does not appear strong or consistent (Hunter et al., 1993).Obesity The relationship of breast cancer to obesity is more complex but associated with a two-fold increase in the risk of breast cancer in postmenopausal women (Morimoto et al., 2002). American Cancer Society study showed that, over weighted women (BMI>25) are 1.3 to 2.1 times to die from breast cancer compared to normal women weight (BMI=18.5-24.9) (Calle et al., 2003). Alcohol consumption Alcohol is consistently associated with increased breast cancer by elevating the estrogen and androgen levels (Hamajima et al., 2002). The National Institute on Alcohol Abuse and Alcoholism (NIAAAA) concluded that, chronic alcohol consumption has been associated with 10% increase in breast cancer (Friedenreich, 1993). Genes Two genes, BRCA1 and BRCA2, have been linked to the rare familial form of breast cancer. Women in families expressing mutations in these genes have a much higher risk of developing breast cancer than women who do not. Not all people who inherit mutations in these genes will develop breast cancer. Together with Li-Fraumeni Syndrome (p53 mutations), these genetic aberrations determine around 5% of all breast cancer cases, suggesting that the remainder is sporadic. Recently it was found that newly discovered gene called BARDI, if exists in combination with BRCA2, gene may increase the12 risk of breast cancer to as much as 80 percent. Genetic counseling and genetic testing should be considered for families who may carry a hereditary form of cancer. Radiation A doubling of breast cancer risk was observed among teenage girls exposed to radiation during the Second World War. Ionizing radiation also increased risk later in life, particularly when exposed rapidly during rapid breast cancer formation stage (Ron, 1998). Polycyclic Aromatic Hydrocarbons (PAH) These are mainly derived from the cigarette smoke, car exhaust and fumes. PAH reacts with the base pairs of cellular DNA inflicting changes that finally lead to cancer. The PAH compounds produce cancer at the site of application in the target organ. The type of cancer produced varies with the route of administration. The PAH compounds are metabolized by cytochrome P4so dependent mixed function oxidase, to give electrophilic epoxide that, either gets detoxified or binds covalently to DNA. The resulting abnormal DNA induces cell division and finally produces tumor. The bay region theory of polycyclic aromatic hydrocarbon carcinogenesis predicts that 7, 12-Dimethylbenz(a)anthracene (DMBA) will be activated to DNA binding forms via the formation of a diol epoxide in the angular benzo ring i.e. a ring leading eventually to a reactive benzyl carbonium ion that can subsequently13 react at nucleophilic centers on the DNA and other cellular molecules (Daniel and Joyce, 1983). Animal models have helped demonstrating different classes of chemicals that act as initiators and induce breast cancer. The DMBA- induced rat mammary carcinomas have been shown to arise from the ductal elements of the mammary gland (Chow ef al., 2003). 7, 12-Dimethylbenz(a)- anthracene (DMBA) induced mammary carcinogenesis in rats has been widely used in various cancer studies (Singletary et al., 1989, Lino et al., 1992). This model appears to be the most relevant to the development of human breast cancer, especially in origin. Types of Breast Cancer Breast cancer can be classified into the following major types, based on the tissues, ducts involved and also depending on the nature of tumors formed. (i) Fibroadenoma The most common benign neoplasm of the breast is the fibro adenoma, a tumor composed of epithelial and stromal elements that originates from the terminal ducts. Fibro adenoma generally appears as a solitary, discrete, freely mobile nodule within the breast. It is a smooth, round, firm and rubbery tumor. It is usually found in women between the ages of 20 and 35.ii) Intraductal papilloma Intraductal papilloma is a common tumor that occurs in the lactiferous ducts of the middle aged and older women. It is situated in the large, subareolar ducts, which may be associated with a serous or a bloody nipple discharge. iii) Carcinoma Carcinomas of the breast develope from the ductal epithelium in 90% cases while the remaining 10% originate from the lobular epithelium. The least common is in intraductal papillary epithelium. For a variable period of time, the tumor cells remain confined within the ducts or lobules (non- invasive carcinoma) before they invade the breast stroma (invasive carcinoma), while there are 3 types of non-invasive carcinoma, there is a great variety of histological pattems of invasive carcinoma breast which have clinical correlation and prognostic implications. Different types of the breast carcinoma as proposed in the WHO classification are as given below. A. NON-INVASIVE (INSITU) CARCINOMA In general, non-invasive carcinoma is characterized histologically by the presence of tumor cells within the ducts or lobules without evidence of invasion.(a) Intraductal Carcinoma This Jnsitu tumor arises in the terminal duct lobular unit, greatly distending and distorting the ducts by its growth. The terminal ducts may become markedly enlarged, thereby resembling large ducts. (b) Lobular carcinoma insitu This type of Jnsitu breast cancer also arises in the terminal duct lobular unit. In lobular carcinoma Jnsitu, the cancer cells tend to be smaller and more monotonous than those of the ductal type, with round, regular nuclei and minute nucleoli. (c) Papillary carcinoma insitu Papillary carcinoma Jnsitu is unusual in that it originates in the larger branches of duct system. The tumor is very well differentiated and exhibits a papillary configuration .The cells are typically small and regular. B. INVASIVE CARCINOMA (a) Ductal carcinoma Invasive or infiltrating ductal carcinoma is the most common form of breast cancer. Invasive ductal carcinoma, usually, presents as a hard, fixed mass, which is often referred to as cirrhosis carcinoma.16 (b) Lobular carcinoma It may occur alone or may be mixed with ductal carcinoma. It comprises about 5% of all breast cancers. The cells of the lobular carcinoma tend to form single strands that invade between collagen fibres. (c) Medullary carcinoma It is the third most common type of breast carcinoma. The malignant cells are pleomorphic and grow in solid sheets, forming a blunt margin. There is no gland formation. (d) Colloid carcinoma Colloid carcinoma is an uncommon (2%) variant that tends to occur in older women. The tumor is usually a soft and gelatinous mass with well- demarcated borders. (e) Tubular carcinoma Tubular carcinoma is a rare form of invasive breast cancer. It is composed of randomly arranged, infiltration, well formed small ducts that consist of one or two layers of small, regular cells. (f) Adenoid cystic carcinoma Adenoid cystic carcinoma is a unique histological pattern of breast cancer with excellent prognosis. Histologically, there is stromal invasion by islands of cells having characteristic cribriform (fenestrated) appearance.(g) Secretory (Juvenile) carcinoma Secretory carcinoma is found more frequently in children. The tumor is generally circumscribed which, on histological examination shows abundant intra and extra cellular PAS positive clear spaces due to secretory activity of tumor cells. (b) Inflammatory carcinoma The term has been used for breast cancers in which there is redness, oedema, tenderness and rapid enlargement. Inflammatory carcinoma is associated with extensive invasion of dermal lymphatics and has a dismal prognosis. (i) Carcinoma with metaplasia Rarely, invasive ductal carcinomas may have various types of metaplastic alterations such as squamous metaplasia cartilaginous and asseous metaplasia or their combinations. STAGES OF BREAST CANCER Cancer of the breast occurs more often in left breast than the right and its bilateral in about 4% cases. Anatomically, upper-outer quadrant is the site of tumor in half the breast cancers, followed in frequency by the central portion, and equally in the remaining both lower and the upper inner quadrants. Breast Cancer can be presented in 4 stages depending on the duration and severity of the disease. The American Joint Committee (AJC) on18 tancer staging has modified the primary Tumor, Nodal and distant Metastasis (TNM) Staging proposed by Union International for Control of Cancer (UICC). Stage -I Stage - II Stage - IITA Stage - III B Stage -IV Tumor 2cm or less in diameter. No nodal spread. Tumor >2cm in diameter. Regional lymph nodes involved. Tumor 25cm in diameter. Regional lymph nodes involved on same side. Tumor 25cm in diameter. Supraclavicular and infraclavicular lymph nodes involved. Tumor of any size, with or without regional spread, but with distant metastasis. Such a differentiation is essential for clinical diagnosis since the choice of treatment between surgery, radiotherapy and chemotherapy depends on the lesion whether it is benign or malignant and whether the particular histological subtype is sensitive to radiotherapy or chemotherapy. There is a great variation in the sensitivity of different breast tumors to several cytotoxic drugs. Therefore appropriate therapy can be prescribed only when the tumor tissue has been accurately classified (Smyth, 1995).Breast Cancer Diagnostic Methods Carcinoma of the breast is the second most common malignancy in the female population and is the cause of morbidity in women all over the world. The recent developments in medical diagnostic techniques such as Mammography, Ultrasonography, Positron emission tomography and Magnetic resonance are used to detect small cancers, visible circumscribed lesion or palpable masses where a cyst is in the differential diagnosis (Jackson, 1990), and to evaluate solid breast masses (Schor and Schor, 1983), breast carcinoma and ductal carcinoma (Boetes et al., 1994). Mammography may help to identify the breast masses, Ultrasonography can find out whether the lump is solid or fluid. In addition to this other methods are available to diagnose the breast cancer. Other types Needle Aspiration Needle biopsies of the mass that yield fluid indicates a cyst. Needle biopsy It removes cells directly from the mass for evaluation. Surgical biopsy It removes a portion of the mass for further evaluation. Incisional biopsy It involves surgical removal of a portion of the mass for evaluation. It enables one to decrease 30% in breast Cancer mortality Shapiro et al.,, 1988). Its improved imaging enables the detection of small cancer (Ferg, 1979).20 Symptoms of Breast Cancer Unfortunately, the early stages of breast cancer may not have any symptoms. This is why it is important to follow screening recommendations. As a tumor grows in size, it can produce a variety of symptoms including: 1. Lump or thickening in the breast of under arm. 2. Change in size or shape of the breast. 3. Nipple discharge or nipple turning inward. 4. Redness or scaling of the skin or nipple. 5. Ridges or pitting of the breast skin. Treatment for Breast Cancer There are different types of treatment for breast cancer used worldwide, they are as follows: Surgery Almost all women with breast cancer will have some type of surgery in the course of their treatment. The purpose of surgery is to remove as much of the cancer as possible, and there are many different ways that the surgery can be carried out. Some women will be candidates for what is called Breast Conservation Therapy (BCT). In BCT, surgeons perform a lumpectomy, which means they remove the entire breast. BCT always needs to be combined with radiation therapy to make it an option for treating breast cancer. Some patients will have a sentinel lymph node biopsy procedure to determine if a formal lymph node dissection is required. Sometimes, they21 remove a larger part (but not the whole breast), and this is called a segmental or partial mastectomy. In early stages of cancer (like stages I & II) BCT is as effective as removal of the entire breast via mastectomy. More advanced breast cancers are usually treated with a modified radical mastectomy, which means removing the entire breast dissecting the lymph nodes under the arm. Radiation therapy Breast cancer commonly receives radiation therapy. Radiation therapy uses high-energy rays (similar to x-rays) to kill cancer cells. It comes from an external source. Radiation therapy is used in all patients who receive breast conservation therapy (BCT).It is also recommended for patients after a mastectomy who had large tumors, lymph node involvement, or close/positive margins after the surgery .Radiation is important in reducing the risk of local occurrence and is often offered in more advanced cases to kill tumor cells that may be living in lymph nodes. Hormonal therapy The pathologist examines the tumor whether they express estrogen and progesterone receptors. Patients whose tumor expresses estrogen receptors are candidates for therapy with an estrogen-blocking drug called Tamoxifen. This drug has been shown to drastically reduce the risk of recurrence if the tumor expresses estrogen receptors. However there are side effects commonly associated with Tamoxifen including weight gain, hot flashes and vaginal discharge that may bother patients. There are also very uncommon side effects like blood clots, strokes and uterine cancer.22 Biological therapy The pathologist also examines tumor for the presence of HER-2/neu over expressions. HER —2/neu is a receptor that some breast cancers express. Its expression shows a higher chance of having the tumor recurrence after surgery. A compound called Herceptin or Trastuzumab is a substance that blocks their receptor and helps to stop the breast cancer from growing. Chemotherapy Despite the fact that the tumors are removed by surgery, there may be microscopic cancer cells that might have spread to distant sites in the body .In order to decrease a patients risk of recurrence many breast cancer patients are offered chemotherapy. Chemotherapy is the use of anti-cancer drugs that go throughout the body. The higher the stages of cancer, the more important it is that they receive chemotherapy; however, even stage-I patients may benefit from chemotherapy in certain cases. In early stage patients, the risk of recurrence may be small, and then the benefits of the chemotherapy are even lesser. Chemotherapy can be given both before and after surgery. Neo- adjuvant chemotherapy is used to shrink the size of a tumor prior to surgery. Adjuvant chemotherapy is given after surgery to reduce the risk of recurrence. There are several different chemotherapy regimens that may be used. The determination of the appropriate regimen depends on many factors23 including the character of the tumor, lymph node status, and the age and health of the patient. Possible chemotherapy regimens include: © CMF: e FAC: e TAC: e FEC: e FEC e TC: cyclophosphamide, methotrexate and 5- fluorouracil 5-fluorouracil, doxorubicin, cyclophosphamide doxorubicin and cyclophosphamide AC with paclitaxel administered after the AC docetaxel, doxorubicin, and cyclophosphamide 5-fluorouracil, epirubucin and cyclophosphamide for 6 cycles for three cycles followed by docetaxel for three cycles Dose dense doxorubicin and cyclophosphamide followed by paclitaxel Taxotere (docetaxel) and cyclophosphamide Side effects of Chemotherapy Generally all chemotherapy drugs have side effects. Most of the anti cancer drugs act by inhibiting DNA synthesis or some other process in the cell growth cycle. It is a known fact that anticancer drugs generally affect24 rapidly dividing cells, other non-cancerous cells (blood cells, dividing cells, hair root, cells lining the digestive tract) may also be affected which lead to side effects in individuals. Side effects may include loss of hair, poor appetite, Nausea or vomiting, diarrhea and mouth or lip sores. The commonest side effects are sickness, diarrhea, hair loss or thinning, feeling tired and sore mouth or mouth ulcers. Alternative medicine and medicinal plants Plants are the principle means of therapy in traditional medicine. Today in many countries modern medicine has replaced plants with many synthetic substances but almost 30% of pharmaceutical investigations in medicinal plants have provided important advances for the therapeutic approach to several pathological conditions in this decade. Plant materials have been used effectively for the treatment of human diseases since ancient times. Every country in the world has enlisted various indigenous herbal temedies according to the disease and human requirements (Dhuley, 1999). Ayurveda, an indigenous system of medicine in India, has a long tradition of treating liver disorders with plant drugs. Ayurveda (Ayur-life; Veda- knowledge), is the knowledge of healthy living and is not merely confined to the treatment of illness. Ayurvedic medicine is largely based on herbal preparations and has specific diagnostic and therapeutic principles (Lee, 1993). About 2000 plants of therapeutic value are mentioned in Ayurveda, Siddha, and Unani systems of medicine. Out of these, quite a large number of plants have been claimed to possess anticancer activity. Herbs or medicinal plants used in the traditional system of medicine contain25 biologically active secondary metabolites, which are not sufficiently screened scientifically for their clinical efficiency. Traditional system of medicine, which is the only accessible health care system for most of the population in tural areas and it should be scientifically evaluated so as to improve the clinical efficiency and to ascertain the safety of medicinal plants. Herbal drugs are being used by about 80% of the world population mainly in the developing countries as primary health care. The chemical constituents present in them are a part of the physiological function of the living flora and hence they are believed to have better compatibility with the human body. Also these drugs are made from renewable resources by ecofriendly processes. In recent years, considerable interest is shown in natural products having antioxidant activity to treat cancer diseases. One of the areas of great deal of attention is the possible use of antioxidant supplement in the prevention of oxidative damage in cells during infections or due to toxicants (Sree priya and Devaki, 2001). Several antioxidant plants have been identified and used as protective agents against oxidative stress induced in diseased conditions (Ohta etal., 2002). Herbal medicines are now in great demand in the developing world for primary health care not only because they are inexpensive but also for then better cultural acceptability, better compatibility with the human body and minimal adverse effects. Medicinal plants are one of the important oldest sources of pharmacologically active compounds and used for the treatment of various diseases.26 Hence the search for an effective anticancer drug still continues. For the above mentioned reasons, it is necessary to take steps to evaluate to develop, to validate and to promote such natural medicines with standard of safety and efficiency which will certainly revitalize the treatment for cancer. PLANT Achyranthes aspera Achyranthes aspera L. belonging to family Amaranthacae is found throughout India in all plains districts and also in other tropical and sub- tropical regions of the world. Classical & common name English : Prickly Chaff Flower Tamil : Nayuruvi, Chirukadaladai. Ayurvedic : Apaamaarga, Churchitaa, Shikhari, Shaikharika, Adahshalya, Mayura, Mayuraka. Unani : Atkum (Arabic), Chirchitaa,Latjeeraaa. Siddha : Nayuruvi. Description Herbaceous plant, about 1m to 2m hight. Stems erect, pubescent, swollen at the nodes, Leaves opposite, short petiole, margins undulate.27 Flowers numerous, stiffly defected against the pubescent vachis in an elongated terminal spike, 20-30 cm long. Urticle oblong, cylindrical enclosed in the hardened perianth, brown, seeds oblong-ovoid. Parts used The whole plant, especially the roots and seeds Chemical constituents and uses The plant Achyranthes aspera L. (Amaranthaceae) is an indigenous medicinal plant of Asia, South America, and Africa that is commonly used by traditional healers for the treatment of fever, especially malarial fever, dysentery, asthma, hypertension and diabetes (Girach and Khan 1992,Tang and Eisenbrand, 1992, Bhom and Liersh, 1992). An extract of the whole plant is described to have diuretic properties and also used for pneumonia. The dried herb is used to treat children for colic and also as an astringent in gonorrhea treatment (Misra et a/., 1991). The roots of Achyranthes aspera are reported to have application in infantile diarrhea and cold (Borthakur and Goswami, 1995) while dry leaves are employed against asthma (Singh, 1995). The seeds are regarded as having emetic and hydrophobic properties (Batta and Rangaswami, 1973). Leaf extracts are reported to possess hypoglycemic, thyroid-stimulating, antiperoxidative activities and anticancer activity. It is reported that Achyranthes aspera contains two oleanolic acid- based saponins (Seshadri et al, 1981). Amino acids and oleanolic acid (Borthakur and Goswami, 1995) have been isolated from the unripe fruits and28 seeds. The occurrence of oleanolic acid has been reported from the roots. Leaves and stems also contain ecdysterone. Alkaloids of the betaine type or betalaine were identified in the leaves and roots of Achyranthes aspera (Bhom and Liersh, 1992). DMBA- induced breast cancer (In-vivo) model A Neoplasm is characterized by an uncontrolled, irreversible, independent, autonomous, and abnormal over growth of tissue (Dev and Dev., 1979). In response to cellular stress caused by carcinogens such as 7, 12-Dimethyl benz (a)anthracene (DMBA) results in mutation of BRAC 1, BRAC 2 genes , P53 protein and tums off transcription factors, thus leading to initiation, promotion and progression of breast cancer (Medina et al., 2002). Several tonnes of polycyclic aromatic hydrocarbons are spilled into the environment every year and these drastically affect the normal life of the living system. Sometimes these effects are stable because of the accumulation of the metabolites from one tropic level to another. It is a well known fact that xenobiotics enter into the environment and cause various disorders (Vijayavel et al., 2005). World Health Organization (WHO) has sent a warning signal that “an increased xenobiotic or environmental pollutant is responsible for the toxicities in human population world wide” (Murray et al., .2001). Polycyclic aromatic hydrocarbons (PAH) are common environmental pollutants that come from incomplete combustion of organic materials, fossil fuel used by motor vehicles, cigarettes, residential heating units and power plants, (Bostrom, 2002. IARC. 1983. U.S. Environmental Protection Agency; 1990.)29 - Polycyclic aromatic hydrocarbon (DMBA) acts as an ultimate carcinogen, mutagen or developmental toxicant. In the metabolism of xenobiotics, cytochrome Pyso or monooxygenases perform an important function in order to eliminate them from our physiological system (Ziegler, 1993), Phase I and phase II reactions convert biologically inactive compounds into active or toxic metabolites. (Williams ef al., 2000). PAH have a capacity to bind with DNA molecules and produce DNA reactive metabolites which are capable of producing cancer on the target organ. (Yuspa, 1986). PAH are metabolized by cytochrome P4sy dependent mixed function oxidase to form epoxide and it is capable of binding covalently to DNA molecule. Thus formation of a diol epoxide in the benzo ring, leading to formation of reactive benzyl carbonium ion that can subsequently react at the DNA and cellular molecules, leads to development of cancer. (Hecht 2002) Moreover DMBA is highly lipophilic in character and easily persists in the adipose tissue of mammary gland and favour the initiation of cancer. (Daniel and Joyce 1983; Singletary et al., 1990). In view of this approach, the present investigation was designed to evaluate the anticancer effects of Achyranthes aspera methanolic extract in experimental breast cancer in rats. Objectives The following parameters were analyzed in control and experimental groups.30 In vitro antioxidant study of the plant extract Effect of Achyranthes aspera on human breast cancer cell line (MCF-7) - In vitro study > > Cytotoxicity study (MTT assay) Light microscopic studies Effect of Achyranthes aspera on DMBA induced breast cancer in experimental animals — In vivo study > ee oe oe pee oe ae ee ae Determination of body weight and tumor weight of the experimental animals. Estimation of total proteins. Estimation of DNA and RNA levels. The activities of marker enzymes. Estimation of lipid peroxidation. Estimation of enzymic and non-enzymic antioxidant status. Determination of TCA cycle enzyme levels. Determination of the activities of membrane bound ATPase . Estimation of biotransformation enzyme system which includes cytochrome P45, cytochrome bs, NADPH cytochrome C reductase, glutathione-s- transferase, UDP- glucuronyl transferase. Agarose gel electrophoresis was performed to investigate the DNA fragmentation. Comet assay was performed. Histopathogical changes were observed for the confirmation of cancer development.