presentation on cancer cell lines with respect to breast cancer

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Cell Lines used for Cancer studies with special reference to Breast Cancer Prepared by:- Ankit Goel M.pharm (PB)

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Cancer involves the abnormal multiplication and spread of cells in the body. It is usually caused by mutations in somatic cell genes that regulate cell growth. Almost every tissue in the body can produce cancer or some even generate many different types of cancer. However, cancer mostly occurs in cells that divide and reproduce more than other cells. Cancer cells also have the ability to migrate from one site in the body where they began and invade other tissues to form tumors at other sites inside the body. This is called metastasis. Many things are known to increase the risk of cancer including tobacco use, certain infections, radiations, lack of physical activity, obesity and environmental pollutant. Approximately 5-10 % cancer are entirely hereditary. What is Cancer?

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Types of cancer Benign Tumors It lacks the ability to metastasize. They are mild and non-progressive disease . Though many kinds of benign tumors are harmless to human health but it still produce negative health effects . Benign tumors typically are surrounded by an outer surface (fibrous sheath) that inhibits their ability to behave in a malignant manner Malignant Tumor Proliferate rapidly, invading neighbor tissues. Can metastasize or spread to other sites of the body Are named using the conventions of tissue, cell type and origin.

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Common forms of CANCER

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CANCER cell lines Cancer is one of the most alarming disease of these days. Today, one out of ten people can be diagnosed with cancer. Carcinogens(cancer causing agents) can be present any where(i.e. soil, water, food, chemicals, sunlight etc.). For the treatment of cancer, scientist are trying hard to determine the cause(carcinogens) and cure for the different forms of cancer prevailing all over the world. These studies are carried on cancer cells(derived from various organisms), which are grown in artificial conditions(i.e. outside the body), by a technique commonly called as ANIMAL CELL CULTURE(ACC). More than 200 different types of cancer prevail in today’s world and around 947 cancer cell lines have been established to study the physiological and pathological aspects of these cancers.

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Types of cell lines Cell lines Meaning Organism Origin Morphology HeLa Henrietta Lacks Human Cervical cancer Epithelium VCaP Vertebra prostate cancer Human Metastatic prostate cancer Epithelial BHK-21 Baby hamster kidney fibroblast cells Hamster Kidney Fibroblast CHO Chinese hamster ovary Hamster Ovary Epithelium MDCK II Madin Darby canine kidney Dog Kidney Epithelium HL-60 Human leukemia Human Myeloblast Blood cells HEK-293 Human embryonic kidney Human Kidney (embryonic) Epithelium HUVEC Human umbilical vein endothelial cell Human Umbilical vein endothelium Epithelia Jurkat yurkat Human T cell leukemia white blood cells LNCaP Lymph node cancer of the prostate Human Prostatic adenocarcinoma Epithelial RBL cells Rat Basophilic Leukaemia Rat Leukaemia Basophil cell

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A HeLa cell , is a cell type in an immortal cell line used in scientific research. It is the oldest and most commonly used human cell line. The line was derived from cervical cancer cells taken on February 8, 1951 from Henrietta Lacks. HeLa cells were used by Jonas Salk to test the first polio vaccine in the 1950s . The cell line was found to be remarkably durable and prolific as illustrated by its contamination of many other cell lines used in research HeLa HeLa cells Scientific classification Kingdom: incertae sedis Phylum: incertae sedis Class: incertae sedis Order: incertae sedis Family: Helacytidae Genus: Helacyton Species: H. gartleri Binomial name Helacyton gartleri Leigh Van Valen

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The HL-60 ( Human promyelocytic leukemia cells )  cell line was derived from a 36-year-old woman with acute promyelocytic leukemia at the National Cancer Institute. HL-60 cell model was used to study the effect of DNA topoisomerase ( topo ) IIα and IIβ on differentiation and apoptosis of cells. It is especially useful in dielectrophoresis studies , which require an aqueous environment with suspended and round cells. HL-60

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LNCaP LNCap stands for Lymph node cancer of the prostate cell lines. LNCaP cells are androgen-sensitive human prostate adenocarcinoma cells derived from the left supraclavicular lymph node metastasis from a 50-year-old caucasian male in 1977. LNCaP cells are a cell line of human cells commonly used in the field of oncology.

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Saos-2 cells Saos-2 ("Sarcoma osteogenic ") is a non-transformed cell line derived from the primary osteosarcoma of an 11-year-old Caucasian girl in 1973 by Fogh . In 1987 Rodan determined that Saos-2 cells "possess several osteoblastic features and could be useful as a permanent line of human osteoblast-like cells and as a source of bone-related molecules ." The latter point is described particularly as "the ability of Saos-2 cells to deposit a mineralization-competent extracellular matrix ", which makes these cells a valuable model for studying events associated with the late osteoblastic differentiation stage in human cells.

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HUVEC The HUVEC cell line was derived and established from human umbilical vein endothelial cells ( HUVEC) . It is used in the study of the interior of the umbilical cord . It may play a role in angiogenesis, or the formation of blood vessels from pre-existing vessels, which supply cancer growths with nutrients . HUVEC systems are commonly used for physiological and pharmacological investigations, such as macromolecule transport, fibrinolysis, and blood coagulation. HUVEC are normally responsive to cytokine stimulation in the expression of cell adhesion molecules.

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Human Embryonic Kidney 293(HEK) cells , are a specific cell line originally derived from human embryonic kidney cells. HEK 293 cells are very easy to grow and transfect very readily. They are used by the biotechnologist to produce therapeutic proteins and viruses for gene therapy. HEK Cells

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BHK-21 Baby Hamster Kidney(BHK) fibroblasts are an adhesive cells lines derived in 1961 by I. A. Macpherson and M. G. P. Stoker . The BHK-21 cells are useful for transformations and for stable and temporary transfections.

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Chinese hamster ovary ( CHO ) cells are a cell line derived from the ovary of the Chinese hamster by Dr. Puck's laboratory in the late 1960's. They were introduced in the 1960s, are grown as a cultured monolayer and require the amino acid proline in their culture medium. CHO cells are used in studies of genetics, toxicity screening, nutrition and gene expression, particularly to express recombinant proteins. Today , CHO cells are the most commonly used mammalian hosts for industrial production of recombinant protein therapeutics . CHO Cell

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The Jurkat cell line (originally called JM) was established in the late 1970s from the peripheral blood of a 14 year old boy with T cell leukemia . Jurkat cells are an immortalized line of T lymphocyte cells that are used to study acute T cell leukemia , T cell signaling , and the expression of various chemokine receptors susceptible to viral entry(HIV). Jurkat cells are also useful in science because of their ability to produce interlukin 2 . They are use to determine the mechanism of differential susceptibility of cancers to drugs and radiation . JURKAT Cells

Breast Cancer:

B reast cancer cells Breast Cancer Breast cancer is a malignant tumor that starts in the cells of the breast. A malignant tumor is a group of cancer cells that can grow into surrounding tissues or spread to distant areas of the body . Breast Cancer occurs when a mutation takes place in the cells that line the lobules that manufacture milk or more commonly in the ducts that carry it to the nipple. It is fairly rare for cancers to form in the fat or non-glandular tissues of the breast. The area around the center of the breast is where most cancers occur .

Signs and Symptoms:

Signs and Symptoms Lumps T hickening different from the other breast tissue O ne breast becoming larger or lower A nipple changing position or shape or becoming inverted S kin puckering or dimpling A rash on or around a nipple D ischarge from nipple C onstant pain in part of the breast or armpit, and swelling beneath the armpit or around the collarbone . Another reported symptom is Paget’s disease of breast.

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Risk factors The primary risk factors for breast cancer are female sex and older age . Other potential risk factors include: lack of childbearing or breastfeeding, higher hormone levels , diet and obesity. Lifestyle Smoking tobacco , those who are long term smokers, the risk is increased from 35 % to 50 % . Use of oral contraceptives results in development of premenopausal breast cancer. Intake of high fat diet & alcohol also increase the risk of Breast Cancer. Dietary iodine deficiency may also play a role . A number of chemicals have also been linked including polychlorinated biphenyls, polycyclic aromatic hydrocarbons, organic solvents and a number of pesticides . Other risk factors include radiation and obesity .

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Genetics Genetics is believed to be the cause of 5–10% of all cases, due to the BRCA1 and BRCA2 gene mutation . In those with zero, one or two affected relatives the risk of breast cancer before the age of 80 is 7.8%, 13.3%, and 21.1% with a subsequent mortality from the disease of 2.3%, 4.2%, and 7.6% respectively . Medical conditions Certain breast changes: atypical hyperplasia and lobular carcinoma in situ found in benign breast conditions such as fibrocystic breast changes are correlated with an increased breast cancer risk

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Types of Breast Cancer In situ Breast Cancer Infiltrating Breast Cancer Infiltrating Ductal Carcinoma nfiltrating L obular Carcinoma Cystosarcoma Phyllode Inflammatory Cancer Breast Cancer during Pregnancy Paget’s Disease

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Clinical Stages of Breast Cancer Clinical Staging is determined by considering the size of the original tumor (T), the lymph nodes (L), and metastasis (M ). This is called the TNM Criteria. T = Primary Tumor Tis = carcinoma in situ T1 = less than 2 cm in diameter T2 = between 2 and 5 cm in diameter T3 = more than 5 cm in diameter T4 = any size, but extends to the skin or chest wall N = Regional Lymph nodes N0 = no regional node involvement N1 = metastasis to movable same side axillary nodes N2 = metastasis to fixed same side axillary nodes N3 = metastasis to same side internal mammary nodes M = Distant Metastasis M0 = no distant metastasis M1 = distant metastasis

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Breast Self Examination Mammogram Sonogram Thermography Transillumination Xeromammograpy Computer Axial Tomography(CAT) Scan Magnetic resonance imaging( MRI) Biopsy Detection of Breast Cancer

Treatment of Breast Cancer:

Treatment of Breast Cancer Chemotherapy - Chemotherapy works by destroying cells that are dividing and multiplying all the time . Radiation Therapy - Radiation, at high energy levels, has the ability to destroy what is in its path, including normal and abnormal cells. Surgery - These are of 2 types:- Mastectomy- In this surgical procedure breast, non-protruding breast tissue, the lymph nodes in the armpits and some pectoral muscle are removed. Lumpectomy- In this surgical procedure, the breast is conserved and the tumor is removed . Drugs- There are four types of drugs that are commonly used for the treatment of breast cancer . Alkylating Agents ( Eg .- Cytoxan) Antimetabolites ( Eg .- Methotrexate & 5-fluorouracil) Natural Products ( Eg .- Vincristine and vinblastine) Hormones ( Eg .- Prednisone)

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Breast Cancer cell lines Breast cancer cell lines have been the most widely used models to investigate how proliferation, apoptosis and migration become deregulated during the progression of breast cancer. The use of cell lines has resulted in a wealth of information about the genes and signalling pathways that regulate these processes. There are numerous reasons why human breast cancer cell lines have been the principal models for breast cancer research. Established cell lines are easily propagated, relatively tractable to genetic manipulation and, under well-defined experimental conditions, generally yield reproducible and quantifiable results . Compared to rodent cells, human cells are frequently perceived as having more ‘relevance to human disease’ owing to the fundamental differences between organisms.

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Breast Cancer cell lines Cell lines Meaning Organism Origin Morphology MCF-7 Michigan Cancer Foundation-7 Human Mammary gland Invasive breast ductal carcinoma MDA-MB-231 M.D. Anderson - metastatic breast Human Breast Cancer T-47D Ductal Tumor Human Mammary Gland Epithelial MDA-MB-468 M.D. Anderson - metastatic breast Human Breast Cancer MDA-MB-435 M.D. Anderson - Metastatic Breast Human Breast Melanoma or carcinoma (disputed) SK-BR-3 Sloan-Kettering Human Breast Epithelial MA-CLS-2 Cell line Service Human Mammary gland Epithelial Colo-824 Human breast Squamous carcinoma cells Human Breast Epithelial

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MCF-7 MCF-7 cell line was isolated in 1970 from a 69-year-old Caucasian woman i.e. Frances Mallon. MCF-7 is the acronym of Michigan Cancer Foundation - 7 , referring to the institute in Detroit where the cell line was established in 1973 by Herbert Soule. This cell line have the ability to process estradiol via cytoplasmic estrogen receptors and the capability of forming domes . Tumor necrosis factor alpha (TNF alpha) inhibits the growth of MCF-7 breast cancer cells. Treatment with anti- estrogens can modulate the secretion of insulin-like growth factor binding proteins.

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T47D The T-47 line was isolated in 1979 from a pleural effusion obtained from a 54 year old female patient with an infiltrating ductal carcinoma of the breast by I. Keydar . This differentiated epithelial cell line was found to contain cytoplasmic junctions and receptors to 17 beta estradiol , other steroids and calcitonin. These cells exhibit epithelial morphology and form monolayers in culture. The T47D cell line is also able to grow on agarose , an indicator of transformation and tumorgenicity , and displays a relatively high colony forming efficiency .

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MDA-MB-435 MDA-MB-435( M.D . Anderson - metastatic breast) cells is a spindle shaped strain isolated in 1976 by R . Cailleau from the pleural effusion of a 31 year old female with metastatic ductal adenocarcinoma of the breast . These cells displayed the dispersed pattern (type II) when treated with immunofluorescent stain for tubulin. All currently available stocks of MDA-MB-435 cells are derived from the M14 melanoma cell line and can no longer be considered a model of breast cancer

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MDA-MB-453 cells were derived in 1976 from an effusion of a 48-year old Caucasian woman with metastatic carcinoma of the breast, involving the nodes, brain and both pleural and pericardial cavities. MDA-MB-453 cells express high levels of functional androgen receptor , providing a valuable in vitro model for further studies on androgen regulation of gene expression, and possibly cell proliferation, in breast cancer. MDA-MB-453 cells were use to determine that G-protein-coupled inwardly rectifying potassium (GIRK) are involved in cellular signalling exist in breast cancer cells . MDA-MB-453

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Glodek , Cass, Ph.D., "A History of the Michigan Cancer Foundation, the Beginnings & Growth of Detroit's Anticancer Movement," 1990, page 68, Michigan Cancer Foundation, Detroit Rahbari R, Sheahan T, Modes V, Collier P, Macfarlane C, Badge RM (April 2009). "A novel L1 retrotransposon marker for HeLa cell line identification". BioTechniques 46 (4): 277–84 . Abeloff MD, Wolff AC, Weber BL, et al. Cancer of the Breast. In: Abeloff MD, Armitage JO, Lichter AS, et al, eds. Clinical Oncology. 4th ed. Philadelphia, Pa: Elsevier; 2008 : 1875–1943. American Cancer Society. Breast Cancer Facts and Figures 2011-2012. Atlanta, Ga : American Cancer Society; 2011 . American Cancer Society. Cancer Facts and Figures 2013. Atlanta, Ga : American Cancer Society; 2013 References

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