Effect of Paclitaxel for Breast Cancer Treatment Essay

Effect of paclitaxel along with withnia sominiferia on lactate dehydrogenase enzyme activity changes in 7,12 di methyl benz(a) anthracene induced breast cancer wistar rats

Dr.N.Muninathan^1*, Dr.P. Mohanalakshmi²,Ambareesha Kondam k., Dr. S. Malliga³

1* – Department of Biochemistry, Meenakshi Medical College and Research Institute, Enathur,

Kanchipuram – 631552, Tamil Nadu, India.

2- Department of Biochemistry, Sri Muthukumaran Medical College, Chennai

3- Department of Biochemistry, ESIC Medical College, Chennai

Abstract

Aim: The purpose of this study is to investigate the changes in the levels of lactate dehydogenase enzyme(LDH) activity and efficacy of combination of paclitaxel along with Withnia Sominiferia against breast cancer in experimental animals. Breast cancer is the commonest cancer among women in all developed countries (except Japan) as well as in North Africa, South America, and southeastern and western Asia. While the incidence of breast cancer appears to be increasing, mortality rates are now declining in at least some western countries. Breast cancer ranks third when both the sexes are considered together and is clearly a significant global public health problem. Design/Methods: Breast cancer was induced in rats by 7, 12 Di methyl benz(a) anthracene (DMBA) at the dosage of 20mgs dissolved in 0.5ml sunflower oil and administered into experimental animals for 28 weeks. In this study, we demonstrated that combination of paclitaxel and withania somnifera revert the changes in the rats from lethal dose of DMBA within 30 days. Results: All the isoenzymes LDH₁ – LDH₅ were observed in cancer bearing animals. Expression of these isoenzymes were found be reduced in paclitaxel and Withania somnifera treated animals. Conclusions: The treatment with combination of paclitaxel and withania somnifera effectively reduced LDH enzyme activity levels. So, from the obtained results it is concluded that paclitaxel and withania somnifera is capable of restoring the breast architecture.

Key words: Withania somnifera, DMBA, Paclitaxel, LDH and Breast cancer. Effect of Paclitaxel for Breast Cancer Treatment Essay.

1. Introduction

Breast cancer ranks third when both the sexes are considered together (Parkin, 1999) and is clearly a significant global public health problem.

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There are nearly 8,00,000 new cases of breast cancer worldwide each year .In approximately half of these patients, breast cancer will be the eventual cause of death. Incidence of breast cancer in Indian women is not as high as in western countries (Sinha et al., 2003) .It is the second most common cancer among women in south India. The age standardized rates vary from 22 to 28 per 1, 00,000 women (Sanghvi, 1998). Although the rates appear to be lower than those seen in developed countries, the burden of cancer in India is alarming.

Worldwide incidences of breast cancer continue to rise and geographical variations in breast cancer incidence indicate that environment factors contribute to overall risk (Millikan, 1995). Exposure to environmental carcinogens early in life is thought to be one of the first events in the development of breast cancer. Each year breast cancer is diagnosed in 910,000 women worldwide and 376,000 women die from the disease. Most of these cases are in industrial countries e.g., North America (180,000) and Europe (220,000).

PAH’s an important class of chemical carcinogens that are widespread in the ambient environment due to fossil fuel combustion for energy production, transportation and industry. DMBA, a potent PAH recognized as an initiator of both skin and liver cancer (Masaaki Miyata et al). The covalent binding of DMBA metabolites to DNA has been implicated as a critical step in the initiation phase of cancers.

Paclitaxel (Taxol), a naturally occurring antineoplastic agent has shown great promise in the therapeutic management of certain human solid tumors particularly in metastatic breast cancer and malignancy involves skin, lung and refractory ovaries.Effect of Paclitaxel for Breast Cancer Treatment Essay.  It is the original member of the taxane group of anticancer drugs derived from the bark and needles of the pacific yew tree “Taxus brevifolia”. Paclitaxel’s antitumor activity was discovered in1960’s during a large scale 35,000 plants-screening program sponsored by the National Cancer Institute (NCI), USA.

Withania is widely used in Ayurvedic medicine in India and in Unani and Middle Eastern traditional medicines, where it is highly regarded as a panacea, aphrodisiac, and rejuvenative. Withania sominifera (L). Dunal (Solanaceae) commonly called Ashwagandha (Sanskrit) is an Ayurvedic Indian medicinal plant, which has been widely used as a home remedy for several ailments.(Bhattacharya et al., 1997)

The use of ashwagandha in Ayurvedic medicine extends back more than 3000 to 4000 years (Upton, 2000) (Agarwall et al., 1997). It has been widely extolled as a tonic, especially for emaciation in people of all ages, including babies, and enhances the reproductive function of both men and women. It has also been used for inflammation, especially in antitumor, arthritic and rheumatic conditions, for asthma, and as a major tonic to counteract aging and promote youthful longevity (Dhuley., 1998).

2. Materials And Methods

1.1. Chemicals:

7,12 Dimethyl benz (a) anthracene and Withania somnifera were purchased from Sigma chemical company, USA. All the other chemicals used were of analytical grade.

1.2. Animal care and housing:

Female Wistar rats, 6-8 weeks of age and weighing 150-200g, were used. The animals were procured from Central Animal House Block, Meenakshi Medical College and Research institute, Kanchipuran, Tamil Nadu, India and maintained in a controlled environmental condition of temperature and humidity on alternatively 12 h light/dark cycles. All animals were fed standard pellet diet (Gold Mohor rat feed, Ms.Hindustan Lever Ltd., Mumbai) and water ad libitum. This research work on wistar female rats was sanctioned and approved by the Institutional Animal Ethical Committee (REG NO. 765/03/ca/CPCSEA).

1.3. Experimental Design

The animals were divided in to six groups of 6 animals each. Group I animals served as control, Group II, III, IV, V as animals treated with DMBA (20mg ) per animal in sunflower oil (0.5ml), three times a week for 28 weeks to induce skin cancer. After tumor induction Group III animals were treated with Paclitaxel (33mg/kg b.wt) once in a week for 4 weeks. Group IV animals were treated with Withania somnifera (250µg/animal) for 30 days. Group V animals were treated with both Paclitaxel and Withania somnifera (as in group III and group IV).Effect of Paclitaxel for Breast Cancer Treatment Essay.  These were Group VI Control animals treated with paclitaxel and Withania somnifera for 28weeks plus 30 days.

After the experimental period of 32 weeks, the animals were sacrificed by cervical decapitation.

1.4. Biochemical analysis

The isoenzymes pattern of lactate dehydrogenase was separated by the method of Dietz and Lubrano (1967).

III. Results

Plate 1 depicts the Isoenzyme pattern of lactate dehydrogenase in serum of control and experimental animals. All the isoenzymes LDH₁ – LDH₅ were observed in cancer bearing (group II) animals. Expression of these isoenzymes were found be reduced in paclitaxel (group III) and Withania somnifera (group IV) treated animals. However a much significant reduction in the LDH isoenzyme expression pattern was observed in-group V animals treated with both paclitaxel and Withania somnifera. Effect of Paclitaxel for Breast Cancer Treatment Essay.

Lactate dehydrogenase (LDH) isoenzyme pattern in serum of control and experimental animals

Lane 1:Control

Lane 2:DMBA treated

Lane 3:Paclitaxel treated

Lane 4:Withania somnifera treated

Lane 5:Paclitaxel and Withania somnifera treated cancer bearing animal

Lane 6:Paclitaxel and Withania somnifera treated control animal

IV. Discussion:

Lactate dehydrogenase (LDH) is the most common clinical enzyme used in the cancer patients for prognostic purpose ( Invone et al., 1998). It has an important role in germ cell functions and can predict responses to chemotherapy and the prospects of remission. Human cancer tissues typically exhibit 2-3 fold increases in glycolytic enzymes and LDH activity. Sandhya Mishra et al. (2004) have also reported the increased level of LDH in breast cancer patients.

Anderson and Kovatik (1981) reported greater LDH activity in breast cancer conditions. There was a significant increase in LDH level in serum of cancer bearing Group II animals. This might be due to the membrane disruptions that caused the release of these enzymes from the cancer cells or the overproduction by the tumor cells (Helmes et al., 1998). The elevated LDH activity may also have resulted from differences in the rate of synthesis, degradation or the excretion of the enzymes in the mammary cancer bearing animals. Schwartz (1973) has reported that among the isoenzymes LDH₅ was six times greater than LDH₁ in mammary tumor cells.

Flavanoids have proved to possess antitumor effect on various animal models (Ames et al., 1995). The biological and pharmacological activity of Withania somnifera was associated with phenolic compounds mainly to flavanoids, aromatic acids and esters (Burdock, 1998; De catsro, 2001). Effect of Paclitaxel for Breast Cancer Treatment Essay. Antioxidant activity of flavanoids may also be due to their structural features and its action on membrane (Saija et al., 1995; Mathur et al., 2003 ; Mohan et al., 2006). Paclitaxel being rich in flavanoid content possess antitumor and antiproliferative activities that stabilizes the membrane permeability and reduces the release of LDH.

V. Statistical analysis

For statistical analysis, one way analysis of analysis of Variance (ANOVA) was used, followed by the Newman-Keuls Multiple Comparison test.

VI. Conclusion

From the present study, the effect of Paclitaxel- Withania somnifera combination proved to be effective chemotherapeutic agent against DMBA induced Breast cancer in wistar rats compared to that of paclitaxel or Withania somnifera confirmed analyzing the LDH isoenzymes levels in serum.

Reference

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Breast cancer is characterised by formation of tumor in tissues of breast. Four type and stages of breast cancer have been identified. Disease majorly affects females and incidence rate is increasing day by day. Different forms of treatments are available to treat this disease. Following a particular treatment depend on condition of patient and desired outcome demanded by patient. All current treatment possesses side effects and need to be rectified. New treatment need to be generated focusing on optimizing the efficacy and minimising the side effect. New treatment can be obtained by promoting the research work in cancer. New treatments are rising hope to obtain cancer free world.

Introduction:

Breast cancer is a disease where in which the tissue from breast forms of malignant cancer. Usually a tube carries milk to nipple (duct) and to words gland that that from milk. It is most common for men and women, although in male disease diagnosis is rare. It is a heterogeneous that differs by indusial age group and kind of cell in the tumor.

History:

Breast cancer is oldest form of carcinoma. The oldest description of cancer was done in Egypt in 1600 BC. Written literature shows that there is no treatment available for cancer.

William Stewart Halsted performed mastectomy in Halsted radical mastectomy both breasts are removed along with associated lymph nodes and underlying chest muscles. It can lead to long term and person and suffering, but it prevents cancer from reoccurring.

Till 1970 radical mastectomies was considered as standard but later metastasis that is migration of cancerous cells from region of origin to other parts of body.Effect of Paclitaxel for Breast Cancer Treatment Essay.  This provided First experimental transmission was done by a French surgeon Bernard Peyrilhe realized the he injected extracts of breast cancer in animal.

Janet Lane-Claypon did the first case control study on breast cancer in 1926 which was a comparative study comrising of 500 breast cancer cases and 500 control patients .one variable method was used.

Classification of breast cancer:

Ductal carcinoma: it is non invasive form of the breast which starts with in cell line of breast duct below the nipple and areola. Milk is supplied by the duct.

Lobular carcinoma: it starts in lobes or glands that form milk. 8% of the breast cancer accounts to be lobular.

Inflammatory breast cancer: it is the least common form of breast cancer which progress approximately 1% to 3% to diagnose the breast will appear swollen and inflame it cause by inflammation by nets or sheets.

Paget’s disease: this type of breast cancer appears as skin rashes over nipple. It shows science of itchy, these signs of itching under the skin. This will indicate small ductal carcinoma in suit (disc) Effect of Paclitaxel for Breast Cancer Treatment Essay.

Breast cancer stages:

Stage 0: this can be use to explain abnormal invasive of breast, it can be use in DCIS. These are diagnosed in which abnormal cells lining to the breast duct, the abnormal cell surrounding breast tissue or progress outside the duct. DCIS can progress to invasive breast cancer if it is left untreated

Stage 1: it’s an early type of invasive breast cancer in which cancer cell penetrate breast tissue from the starting point of cancer the size of tumor is not more than 2 cm

Stage 2: in this cancer progress to the lymph node the size of the tumor is between 2 and 5 cm, it does not progress to the lymph node it’s around 5 cm.

Stage 3: it consist of two type stage’s

Stage 3A: in this type of cancer the tumor size is large than 2 centimetres and smaller than 5cm and it spread under arm of the lymph nodes.

Stage 3B: cancer progress to tissue surrounding breast along with skin chest wall, ribs, and lymph nodes, in the wall of the cheats.

Stage 4: it is a distant metastatic cancer. Effect of Paclitaxel for Breast Cancer Treatment Essay. This type of cancer is spread whole part of the body, like bones, liver etc.

Incidence:

How common is breast cancer:

As per the report in 2007, 45,700 women were victim of the breast cancer. While in the same year the men count goes to 277 were diagnosed. Incidence rate of breast cancer in females around by 50% over the last twenty five. Out of ten eight of women age of fifty were sufferings from breast cancer.

Breast cancer rates have increased up to 5% in last ten years.National Health Service (NHS) screening programmes were conducted in that more than 16,000 cases found in UK in 2007/2008. Among that NHS screening programme claim to save 1,000 lives each year. Throughout the world 1.38 million women were treated with the breast cancer.

Breast cancer incidence were highly considerable in western and Europe as per the low rate considerable in Africa and Asia. Europeans union shows cancer diagnosed around 332,000 in the year of 2008. Twelve thousand women and seventy men were died from breast cancer in 2008 in united Kingdome. More than half of seventy years of age are died from breast cancer. Worldwide 448,000 women are died cause of the breast cancer in the year of 2008.

Symptoms:

Major symptoms can be characterised as

Formation of lump or thickening inside or surrounding portion of breast

Alteration in shape and size of breast

In certain cases unusual discharge from nipples may be diagnosed

Breast skin may show phenomena like dimpling or puckering

There can be alteration in size and shape of breast

Breast cancer may be associated with several common disorders like fibro adenoma and mastitis. Care provider must be consulted in regard of appearance of any such symptoms. Over 80% of total cases diagnosed with breast cancer are diagnosed at the time when women feel lump. Mammogram is the tool to detect earliest breast cancer.Effect of Paclitaxel for Breast Cancer Treatment Essay.  Breast cancer can also be indicated by means of lumps that are found in lymph node. Which is present in armpits? Alteration in shape and size of breast can also be indicating that a person is suffering from breast cancer. Pain is another factor which can suggest that a person has became victim of breast cancer.

Age: most of the women above the age of 50 to 60 years are suffering from breast cancer and they diagnosed.

Health history of the perso0n: breast cancer in one of the breast will increase the risk of getting cancer in other breast. It consists of certain types like ductal carcinoma in suit (DCIS), lobular carcinoma in suits (LCIS) increase the risk of cancer. These are seen in biopsy of the breast.

Family history: chances of breast cancer are getting higher from ancestors either from mothers or fathers family. There are chances of ovarian cancer with breast cancer also increase risk.

Risk factors which can be control:

Epically heavy weight women, are in risk of getting breast cancer mainly after menopause. Estrogens are the main source for body’s fat tissue after menopause, when hormones are not secreted from ovaries, estrogens can causes cancer risk

Regular exercise: regular exercise can decrease the cancer risk; daily 25-35 minutes of exercise can reduce cancer.

Alcohol taking: in women’s excess of alcohol taking causes by breast cancer and it reaches its ability to control limits of liver.

Smoking: smoking can causes high risk of breast cancer. Effect of Paclitaxel for Breast Cancer Treatment Essay.

Treatment:

Now a day’s who r suffering from breast cancer have many types of treatment and therapy are available, some of treatment are

Surgery

Radiation therapy

Hormone therapy

Chemo therapy

Targeted therapy

Among this all therapy’s most common therapy is radiation therapy in which we can destroy the cancer cells in breast Systemic therapy are hormonal therapy and targeted therapy in this the blood enters and destroy the control system of the body Stage of cancer mainly depends on,

Surgery:

The treatment which is treated most commonly in breast cancer is surgery these are two types they are:

Breast- sparing surgery

Mastectomy

Breast-sparing surgery: the target of breast sparing surgery is to specifically remove tissue’s surrounding the tumor which may involve removal of the lump or quarter of breast.

Mastectomy: in women’s suffering from mastectomy will remove the whole breast if needed will have suitable treatment in case of lump enlargement in small breast or a lump situated middle of the breast. Effect of Paclitaxel for Breast Cancer Treatment Essay. There are various type of mastectomy are differentiate

Either it remove the breast tissue these breast tissue includes skin and nipple

The other type is radical mastectomy which removes the muscle of the chest wall

Radiation therapy:

Radio therapy is use for the radiations like x-rays; it can destroy the cancer cells were it is treated and the effects go to the normal cells by radiation. These are two types they are

External radio therapy

Internal radio therapy

External radio therapy: these are use in high energies of x-rays like irradiation and cobalt.

These can be destroyed the cancer cell to stop growing or die when the DNA is damage.

Internal radio therapy: in this to treat the cancer by using radioactive metals, in this two types of radioactive therapy are there 1) radioactive implement 2) radioactive liquids

Hormone therapy:

Hormone therapy is based on use of naturally produced chemicals as a treatment approach. Two harmones that are estrogen and progesterone manages various activities inside the body. Hormone therapy focus on shrinkage of cancer area .if further surgery is required then harmone therapy is used before it as it decreases the task of removing whole breast during surgery. Harmone therapy works only in the presence of oestrogen receptors. Tamoxifen and aromatase inhibitor lies in this category. Tamoxifen is known to be an estrogen antagonidst because its structure resembles alot with synthetic oestrogen.Effect of Paclitaxel for Breast Cancer Treatment Essay.  Being structurely similar to estrogen it act by binding to oestrogen receptor and leads to formation of non productive complex so oestrogen receptor count decreases and growth promotion effect of oestrogen get suppressed. Side effect of tamoxifen includes hot flashes, vomiting, and skin rashes.

Chemo therapy and their assessment:

Medications for treatment of breast cancer belong to four categories. First category is of ant metabolites which inhibits action of metabolite as these compounds are similar in structurally to metabolites. Medications belonging to this category are 5 fluorouracil and methothrexate. Second categories of medication are antibiotics like doxorubicin and epirubicin. Third categories of medications are inhibitors of microtubule formation like paclitaxel which belong to taxol group. Forth category of medication is alkylating agents like cyclophosphamide. All these medicatins act differently. Alkylating agents like cyclophosphamide gets activated in liver by cyt p450 enzyme and leads to alkylation of DNA. Antibiotic like doxorubicin acts by intracting with molecular oxygen leading to formation of superoxide and hydrogen peroxide which ultimately leads to single step breakdown of DNA. Taxol group act by binding reversibly to beta tubulin and leads to over stabalization of beta tubulin which is non functionalthis result in cell death of infected cells. All treatment possesses a number of side effects. Cyclophosphamide causes bone marrow depression. Paclitaxel causes neutropenia that is decrease in number of neutrophils. Methothrexate shows renal demage, hepatic function alteration and pulmonary toxicity. Fluoro uracil shows bone marrow depression as major toxicity.

All these medication possess lot of side effect so research work need to be done at a faster pace to obtain a treatment with good therapeutic effect and least side effects. Effect of Paclitaxel for Breast Cancer Treatment Essay.

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Target therapy:

Target therapy has emerged as a new approach now days. It involve use of drug that stabilizes breast cancer as targeted therapy can block action of certain undesired proteins like HER2 that promote growth of breast cancer cells.

Trastuzumab: it is provided intravenously that is through vein. It can be used as mono therapy or in combination with chemotherapy. Side effects related to first treatment include fever, chills, weakness, nausea, vomiting, diarrhoea, headaches, difficulty in breathing, and rashes.

After first treatment side effects become less severe as tolerance develops. Trastuzumab can lead to heart problem, and serious breathing problems. Doctor need to check status of heart and lungs both before and in between of treatment.

Lapatinib: it is orally available medication which is given in combination with chemotherapy Side effects of this are nausea, vomiting, diarrhoea, tiredness, mouth sores, and rashes. It can also lead red, painful hands and feet.

Treatment classification on basis of stage of disease:

First factor need to be considered is size of the tumour with respect to size of breast

Laboratory result for status of breast

Factors related to age as patient is in menopause or not General health of patient.

Conclusion:

Breast cancer can be diagnosed by presence of tumour cells surrounding the breast. This disease primarily affect females .variety of treatments are available .each treatment follow different method to change the state of disease. Side effects of current treatment are severe so new treatment need to be developed which can effectively treat this disorder. Reseach work need to be promoted to obtain a good treatment with least side effect and that can specifically target cancer cells. Effect of Paclitaxel for Breast Cancer Treatment Essay.