Isolation and Characterization of Oral Cancer Cell Membrane Vesicles - Coursework Example

ISOLATION AND CHARACTERIZATION OF ORAL CANCER CELL MEMBRANE VESICLES By of School and Background Information Currently, there are increased cases of cancer developments not just within the United States and Europe, but across the world. Basically, oral cancer- a form of malignancy originating from epithelial neoplasm and affecting the oral cavity greatly hampers normal body functioning. This form of carcinoma affects any part of the oral cavity and is scientifically thought to be predisposed by various factors both genetic and environmental (Tsantoulis, et.al. 2006, 523; American Cancer Society. 2014). Like other form of cancer, OSCC- the most common form of oral cancer can be lethal if not diagnosed and treated early (Marocchio, et.al. 2010, 267). Currently, the oral malignancy accounts for 2-4%, with higher prevalence in India and Pakistan, an observation that is attributed to chronic smoking and alcoholism experienced in the two nations (Markopoulos, A. 2012, 126). According to the US Department of Health and Human Services, prior exposure to neck cancer, sunlight and use of lip bums are some of the predisposing factors for the medical condition. Studies have also shown that the oral cancer strain has undergone genetic evolution, with persons having specific gene-lines such as ADH3 and CYP1A1 standing higher chances of infection (Tsantoulis et al. 2006, 524). Currently, oral cancer ranks eighth among cancer variants globally (Massano et al. 2006). Therefore, oral cancer can be eradicated through positive attitude involving zero-tolerance to alcohol and tobacco abuse, and ensuring personal care aimed at eradicating carcinogenic conditions (National Cancer Institute. 2003, 6-7). Types of membrane vesicles For a long time, a wide number of microvesicles and membrane elements have been observed in body fluids and within the mouth cavity. Recent discoveries have associated most of the vesicles with cancer development (Doormal, et al. 2009, 194). Some of the membrane vesicles that have been of interest in cancer studies include; the exosomes, the apoptic bodies and the microvesicles. In some instances, it has been proposed that the microvesicles and membrane elements act as communication mediators during cancer development (Muralidharan-chari, et al. 2010). Exosomes in cancer development (Source; Henderson, M. and Azorsa, D. 2012) The exosomes are cancer induced membrane vesicles that are currently widely used to monitor cancer development. These vesicles are highly stable hence less dissolved in bio-fluids and are proposed to play a vital role in development and progression of oral cancer. Anatomically, exosomes are up to 100nm in size and are composed of widely varying contents. The enclosing membrane is lipid rich, containing high ratios of ceramidine, sphingomyelin and cholesterol all of which have specialized roles (Henderson, M. and Ozorsa, D. 2012). The membrane-bound vesicles are widely distributed in mammalian body occupying virtually all body fluids. Exosomes secretion is a well modulated process with the ability to occur through two distinct pathways, the voluntary activities of the Golgi apparatus and the induced vesicle production mechanism. The latter technique is more common in cancer patients hence an increased level of exosomes in body fluid can be effectively used as a clinical indicator of carcinoma. In cancer induced states, the massive proliferation and production of exosomes is important in many ways. Firstly, the exosomes portray anti-tumor properties. This property is easily exploited by the body defense mechanisms to establish protective measures against the tumor cell through induction of programmed cell death. This property has been exploited in development of exosomes-based cancer vaccines. However, the rate of proliferation and mutation of cancer cells is often too high to be fully controlled through apoptosis (Henderson, M. and Ozorsa, D. 2012). According to Yang, C and Robbins, P (2011), exosomes contain antigen specific and are hence produced in response to specific antigens. This therefore implies that an exosomes is likely to be rendered ineffective in its anti tumor role should the target antigen undergo mutations, a common mechanism used by carcinogenic cells to evade identification and eventual degradation by body defense mechanism. While the tumor-induced exosomes can be credited for their anti-tumor properties, the exosomes are also used as invasion tools by the cancer cells. As a result, the body defense is weakened making the host to be immune-compromised. This occurs in instances where the tumor-derived exosomes produce death signals such as TRAIL initiating apoptosis of T-lymphocytes. Some exosomes inhibit activity of NK cells thus preventing immune reactions towards cancer cells. Following development of a weakened immune system, the exosomes facilitate tumor invasion of body tissues, and its development into grave cancerous conditions. Moreover, the membrane vessels are central in the transport of genetic machinery, proteins and survival factors necessary for cancer development. In response to tumor-directed drugs, exosomes facilitate drug interference rendering medications ineffective. In laboratory setup, exosomes can be biochemically determined through western blotting. In this case, the antibodies are able to identify special exosomal surface proteins including; Hsp70, CD9, CD 81 and CD63. Apoptotic bodies (Source; Kolluri, 2010) During cancer development, rapid cell proliferation and degradation occurs. Therefore, scientific analysis of programmed cell death can be ideally used as a diagnostic tool for cancer. However, oral cancer cells, contrary to other malignant forms, exhibit less apoptosis making it imperatively difficult to diagnose early stages of cancer development (Jain, et al. 2009). The process of apoptosis is biologically controlled by several factors. Key among these includes the Bcl-proteins, TNF-R, Caspases and Adapter proteins (Jain, et all. 2012). The apoptosis process occurs through a series of cascades, most notably the Apoptotic signal cascade. During the process, extrinsic cell death is initiated by the attachment of the ligation of death signals on the cytoplasmic membrane. This causes functional distress to mitochondria and other vital cellular machinery. As a result, the cell is deprived of critical structure causing death. While normal apoptosis is essential in ensuring homeostasis of physiological processes, abnormal apoptosis can be indicators of disease conditions such as cancer. Cancer induced apoptosis can indicate giant cell granuloma, pyogenic granuloma and inflammatory diseases. This diseases can result from oral irritation, and are often suppressed by females hormones, explaining the reason for increased oral cancer instances are more prominent in males than in females (Hsu, S., Singh, B. and Schuster, G. 2004). The apoptic bodies can be determined through identification of mRNA linked tyrosinase, a malignant variant of the free tyrosinase found in normal cells. Microvesicles (Source: Cornell University College, 2012) In cancer development, intracellular and extracellular communication is vital. As a result, cancer cells often secrete Microvesicles (MVs) as communication signals through which they can influence nearby cells. This influence occurs through paracrine manner, but at times the Microvesicles can fuse with the target normal cells (Nieudwland et al. 2009, 195). During the fusion, the MVs discharge their malignancy-promoting contents into normal cells consequently converting the cells into cancer cells. According to laboratory observations, it has been pointed out that microvesicle secretion increases rapidly during cancer development, but the levels remain relatively low under normal body conditions, and are critical in trans-cellular transfer of active bio-constituents (DSouza-Schorey and Clancy, 2012). While the vesicles have commonly been implicated for spread of diseases, research indications have pointed towards a potential application as anti-cancer agents. According to Cornell University College (2012), the MVs help the cancer cells to spread their influence without having to move. As a result, the institution points out that microvesicle targeting can impair intercellular communication. Consequently, the effect of cancer cells will be highly localized and easier to control through radiation or surgical techniques. To determine cancer induced microvesicles, the physical and biochemical properties of the MVs are determined. For instance, the malignant forms have chromosomal abnormality, abnormally high degradation rate and alarming rates of proliferation (Kawamoto et al., 2012). miRNA MicroRNAs are post transcriptional gene modifiers found in eukaryotic cells. Biosynthetically, miRNAs are generated through action of RNA Polymerase II on intronic regions of human genes. The generated transcript of the RNA is then subjected to post transcriptional modification to produce fully functional microRNAs capable of regulating target genes. In cancer development, miRNA can be vital in down-regulating the rate of expression of cancer genes. As a result, the cancer cells lose their ability to rapidly proliferate and grow. In addition, the miRNAs can induce programmed cell death thus reducing the number of cancer cells. Therefore, it is proposed that effective targeting of the microRNAs to specific cancer machinery can be an effective mechanism in controlling the malignancy (Espinosa and Slack, 2006). Oral fibroblasts There are many subpopulations of oral fibroblasts with highly specialized protective roles. For instance, the fibroblasts are central in the proliferation and re-development of mesenchymal tissues during would-healing process. Immunologically, the oral fibroblasts are involved in the secretion of various cytokines such as IL-1 and HGF which are important in immunological defense against invading antigens (Rodemann, P. and Rennekampff, O. 2011). Through various research ventures, oral fibroblasts can be associated with cancer development. Consequently, research ventures have regularly opted to use the fibroblasts as biomarkers for oral cancer (OSCC). According to san Miguel et al. (2012), the oral epithelial lining is subject to corrosive effects of a wide array of toxic substances. Such substances as hydrogen peroxides and other reactive oxygen species kill epithelial and inflammatory cells which occur in close proximity to oral fibroblasts. Consequently, the oral cavity is bound to develop wounds that may hamper the proliferation and development of oral fibroblasts, which depend on epithelial cells as functional and physical regulators.. This decrease in the number of fibroblasts results into increased susceptibility of the patient to oral infections and cancer (Karagiannis et al., 2012). In some instances, however, cancer cells can develop a close association with oral fibroblasts. The cancer-associated fibroblasts (CAFs) can then interact with body cells to create an environment favorable for cancer development. To facilitate this special role, the CAFs reform the peritumoral stroma, facilitating invasion, proliferation and metastasis by the cancer cells. The mal-formed fibroblasts also ensure the cancer cells are subjected to favorable in-situ environment where they can remain intact and effectively multiply (Karagiannis et al., 2012). References American cancer society, (2014). Oral Cavity and Oropharyngeal Cancer. 1st ed. [ebook] american cancer society, pp.1-68. Available at: http://www.cancer.org/acs/groups/cid/documents/webcontent/003128-pdf.pdf [Accessed 31 Jul. 2014]. Cornell University College, (2012). Stopping the spread of cancer. [online] Cornell University College of Veterinary Medicine. Available at: http://www.vet.cornell.edu/news/alumni/12aug/cargo.cfm [Accessed 1 Aug. 2014]. DSouza-Schorey, C. and Clancy, J. (2012). Tumor-derived microvesicles: shedding light on novel microenvironment modulators and prospective cancer biomarkers. Genes & Development, [online] 26(12), pp.1287-1299. Available at: http://genesdev.cshlp.org/content/26/12/1287.full [Accessed 31 Jul. 2014]. DSouza-Schorey, C. and Clancy, J. (2012). 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