Introduction
The total number of patients with cancer keeps soaring globally with around 14 million cases diagnosed by 2014. Each year, about 8 million people die of cancer-related illnesses globally. There are quite some remedies that have been proposed to mitigate in the cancer menace. This study is focused on cancer treatment by curcumin through tyrosine kinase inhibition. Curcumin is the primary coloring agent found in turmeric, a tuber of the herb Curcuma longa Lin. It has widespread use as a flavoring agent (Aggarwal and Bokyung 87). It is also well known for its medicinal value commonly used to suppress inflammations. It has been associated with anticarcinogenic advantages in fauna, as attested by its capability to hinder both tumor origination initiated by benzopyrene and 7, 12-dimethylben anthracene and tumor advancement induced by phorbol esters, probably by blocking protein kinase (Kunnumakkara et al. 200).
According to Yang and Cheng-Liang, et al. (46), Curcumin is known for its anti-tyrosine kinase property and has been proved to slow down ligand-initiated stimulation of the epidermal growth factor receptor tyrosine phosphorylation with minimal effect on the protein level. The erbB-2/neu (commonly referred to as HER-2) gene encrypts the 185-kDa protein that is known to have tyrosine kinase activity (Chen et al. 278-287). It has been documented that in over 30% of breast cancers, 185-kDa and is the primary contributor to poor prognosis. A Transfecting breast epithelial cell with erbB-2/neu alters these cells, the cancer cell lines transfected with erbB-2/neu exhibit an improved ability to attack and metastasize (Kunnumakkara et al. 201; Karunagaran 117-129.
Animal model experiments have revealed that decreasing p185neu by suppressing the erbB2/neu promoter or by utilizing anti-p185neu antibodies can yield greater results with regards to addressing tumor growth and as well as its distribution (Bharti et al. 3863). An inhibitor of p185neu tyrosine kinase, emodin, equally biasedly depress the progression of cell lines that express high levels of p185neu (Natarajan & John 6707).. Additionally, clinical trials have revealed that humanized anti-p185neu monoclonal antibody is equally valuable. Together, these observations show that by moderating p185neu may be of significant value in the therapy breast cancer (Kunnumakkara et al. 201; Karunagaran 117-129). Considering that curcumin has already been revealed to promote reduced systemic harmfulness both in animal and human studies, this study will look into the impression of curcumin on p185neu tyrosine kinase. In doing so, the study will seek to establish whether p185neu tyrosine kinase is hindered by p185neu tyrosine kinase activity and, more significantly, if it exhaustively attacks the protein itself. The study will take note if curcumin is a suitable agent to dissociate the conjoined p185neu and GRP943 protein; which is regarded as a molecular chaperone that accelerates the weakening of mature p185neu. Curcumin is expected to depress the multiplication of various breast cell lines and favorably down-regulate the colony formation of cell lines overproducing p185neu.
Rationale
It is the researcher’s belief that this research’s findings would shed light into two main areas. First, it would lead to a better understanding of cancer treatment by curcumin through tyrosine kinase inhibition. Enhancing awareness and understanding cancer treatment by curcumin through tyrosine kinase inhibition will promote preparedness of the sick and the caregivers in utilizing medical nutrition therapy in an informed manner. Second, this research has the potential to produce knowledge that can be used by clinical nutrition guideline developers at both national and international level to craft policies related to promoting the use of cancer treatment by curcumin through tyrosine kinase inhibition. Finally, it is my belief that this research would produce shared common knowledge that both patient and healthcare provider and its results can be can use as a basis to medical improvement nutrition in cancer treatment (Korutla et al. 1741; Korutla 770; Shishodia et al. 250 & Anand 134).
The aims of the present thesis are threefold: will look into the effectiveness of curcumin on p185neu tyrosine kinase, and establish whether p185neu tyrosine kinase is hindered by p185neu tyrosine kinase activity and, more significantly, if it exhausts the protein itself and finally take note curcumin will be a suitable agent to dissociate the conjoined p185neu and GRP943 protein,
The following research objectives were formulated:
Objective One: To determine the effectiveness of combined therapy using curcumin on p185neu tyrosine kinase.
Introduction
Most of the actions related with curcumin connect to its use to address acute and chronic inflammation. Experimental (vitro investigations) have revealed that curcumin depresses lipooxygenase and cyclo-oxygenase processes in phorbol 12-myristate 13-acetate (PMA)-initiated inflammation of mice fibroblast cells, xanthine oxygenase processes in NIH3T3 cells, nitric oxide production in RAW264.7 murine macrophages, and reactive oxygen species (ROS) generation in activated rat peritoneal macrophages. Currently, curcumin hampers the release of pro-inflammatory monocyte/macrophage- developed cytokines, incendiary monocyte protein-1 (MIP-1), monocyte chemotactic protein-1 as well as, and tumor necrosis factor-a (TNF- a)] in PMA- or LPS-motivated peripheral blood monocytes. In an exploratory study that was done recently, the researchers found out that oxidative promotion of G proteins in human brain membranes by metabolic prooxidants, homocysteine, and hydrogen peroxide, is likely to be was immensely depressed by curcumin intake. Products made from curcumin were confirmed as suitable inhibitory agents for fat peroxidation in a rat liver microsome readiness and rat brain homogenates, during which curcuminoids showed a more influential antioxidant action than alpha-tocopherol (Plummer et al. 6015; Chaudhary 1-5 & Druker et al. 1035)
Methodology
Obtaining results for this objective will entail an examination of secondary data on curcumin toleration with respect to its bioavailability and then compared to experimental data collected under his watch during the second phase. An example of data that will be reviewed was collected in Taiwan. Data collected from all the patients in phase one will be compared to data in phase two. Here, the researcher will be interested to document how curcumin will be administered as a single daily ingestible dose varying from 500 to 8000 mg/day for five months. He will record the observations to rule out any misconceptions of toxicity.
Expected outcomes of the proposed research
It is expected that the research results will show that curcumin is not toxic to animals and people even when on mega sources even. However, future studies will need to determine the recommended daily allowances for healthy and cancer patients.
Possible difficulties
There is a high likelihood of difficulties linked to the interpretation of literature on curcumin because of the infrequency with which curcumin content is determined and conveyed. There is no doubt that Curcuma longa has curcumin. However, curcumin and curcuminoids have equally been obtained from the other plant species, such as Curcuma aromatics. Additionally, there is an extensive variation of curcumin composition among various commercially processed blends of turmeric. In circumventing such difficulties, the research seeks to read the food labels of different curcumin-containing containers before making any rush judgments.
Objective Two: To establish whether p185neu tyrosine kinase is hindered by p185neu tyrosine kinase activity
Introduction
Human breast cancer cell line, AU-565 will be solicited from a credible Bioscience Laboratory. Other breast cancer cell lines including SKBr3, MDA-MB435, MCF-7 will be will be obtained from the American Type Culture Collection in the nearest station. AU-565 are among the cells that are known to overexpress p185neu, unlike MDA-MB231, MDA-MB435, MCF-7, etc. that express the an average level of p185neu. All of the cells will be grown in ambient conditions, that is, under the supplementation with 9-10% of immature bovine serum and gentamicin (50 mg/ml). The growing medium will be kept to be in a humid condition in an incubator with normal human body temperature, under 5% Carbin (IV) Oxide (Hong et al. 1884; Duvoix 189).
Experiment
During the experiment, the cells will be washed thrice with PBS and then lysed in lysis buffer with a solution of known concentration. The protein composition will be assessed against a normalized measure of a Bio-Rad protein assay equipment. After this, 500 μg of protein will be immunoprecipitated using a monoclonal anti-p185neu antibody on a protein A-Sepharose. The immune complexes will be cleaned, then stay separated by use of 7.5% SDS-PAGE, and transported to nitrocellulose filter paper to ease drying. Non-specific binding on the nitrocellulose filter paper will remain minimal with a separating buffer comprising of nonfat dry milk and Tween 20 (Moulder et al 8888; Singh et al. 24996). The treated filter paper will be nursed with primary antibodies and then incubated with the horseradish peroxidase-conjugated goat antimice antibodies. The bands will be pictured with the boosted chemiluminescence lense. The intensity of the bands will be scanned and computed using special NIH computer software (Dorai and Bharat 130; Shishodia et al. 1270).
Expected outcomes of the proposed research
It is anticipated that this study arrives at the conclusion that curcumin hinders p185neu tyrosine kinase action in vitro and diminishes mature p185neuin vivo. After ten hours of curcumin treatment, the p185neu at the plasma membrane is expected to almost undetectable. Curcumin will break down the multiplex between p185neu and GRP94. However, future studies need to establish that GRP94 is a close associate of p185neu and tends to stabilize it and that this dissociation heralds the exhaustion of mature p185neu at the plasma membrane level.
Challenge anticipated
This is a new area that keeps and it keeps evolving. It is feared that there may not be updated data to back up the findings that will be witnessed herein. Among the currently known tyrosine kinase inhibitors, curcumin is the only naturally occurring substance known to hamper tyrosine kinase action and even downregulate the tyrosine kinase protein itself. To counter that, the researcher will compare results with that of mice.
Literature Review
Curcumin and Induced of apoptotic mechanisms
The property of curcumin that initiates apoptosis in tumor cells without cytotoxic consequences on healthy cells augments the appreciation of the anti-cancer significance of curcumin. This herb is said to be an effective inducer of apoptosis in various cell lines such as HL-60, K562, MCF-7, and HeLa. Curcumin also promotes apoptosis in scleroderma lung fibroblasts (SLF) with no effect of normal lung fibroblasts. This property appears to stem from the fact that there is a staggering level of protein kinase (PK) C3 in SLF, producing moderate amounts of glutathione S-transferase. Shishodia (243) suggested that the inducing of Caki, programmed cell death is actuated by Akt dephosphorylation (Chaudhary 1-5), Bcl-2, Bcl-XL and inhibitor of apoptosis (IAP) protein inhibition (Druker et al. 1035), and cytochrome release and caspase 3 activations (Aggarwal and Bokyung 87). These observations agree with results by Plummer et al. (1615) who examined activations in melanoma and HL-60 cells. Korutla et al. 1741; Korutla 770; Shishodia et al. 250 & Anand 134). Korutla 770 described that curcumin induces caspases during which p53 remains unchanged. Nonetheless, the death receptor pathway is activated through Fas in a Fas-Ligand independent way (Plummer et al. 6015; Chaudhary 1-5 & Druker et al. 1035). Shishodia et al. 250 ascertained that the role of Bcl-2 and Bcl-XL inhibition by preventing curcumin-initiated apoptosis after overexpressing these two essential proteins. Shishodia et al. (250) examined cytochrome c release and caspase 3 activations. These authors were opposite, apoptosis in Jurkat cells is delineated to be caspase three independent, its activation being halted by an increase of glutathione (GSH) amounts (Shishodia et al. 250). Caspase activation by curcumin was depicted to be obstructed by heat shock protein, which does hardly influences cytochrome c release (Plummer et al. 6015 & Druker et al. 1035). However, Chaudhary 1-5 confirmed that curcumin hindered proteasome activity in mice, theoretically resulting in induction of apoptosis through caspase activation (Korutla 770; Shishodia et al. 250 & Anand 134). Regrettably, in chosen pathologies, curcumin can deter chemotherapeutic benefits by reducing camptothecin (Plummer et al. 6015), mechlorethamine- or doxorubicin-induced apoptosis in breast cancer cells (Chaudhary 1-5). Curcumin showed antioxidant properties and inhibited both JNK activation and mitochondrial release of cytochrome c in a concentration-dependent manner (Plummer et al. 6015). Nonetheless, a connection of curcumin with other antitumor drugs should be thoroughly evaluated in breast cancer (Korutla 770; Shishodia et al. 250 & Anand 134). Even a restriction of exposure of patients to curcumin-rich foods should be given a consideration (Dorai and Bharat 130; Shishodia et al. 1270).
Curcuma and its interactions with Cell Signaling Proteins
Lates targeted therapies such as celecoxib, avastin, erbitux (Druker et al. 1035), enbrel, herceptin, gleevec, and avastin. It also depresses other regions such as transcription factors (Plummer et al. 6015), enzymes, growth factors and its receptors (Shishodia et al. 250) & Anand 134). Celecoxib, is a non-steroidal anti-inflammatory medication that biasedly inhibit COX-2 Shishodia et al. 250 & Anand 134), avastin, a monoclonal antibody to fight VEGF; Remicade (infliximab), a monoclonal antibody against TNF (Plummer et al. 6015; Chaudhary 1-5 & Druker et al. 1035). This performs crucial roles in Humira, humanized monoclonal antibody against TNF; Enbrel, a recombinant human soluble TNFR fusion protein; Herceptin, humanized monoclonal antibody against HER-2; Gleevac a many tyrosine kinases inhibitors working together (Druker et al. 1031).
According to (Plummer et al. (6020), human alpha1-acid glycoprotein; aryl hydrocarbon receptor; autophosphorylation-initiated protein kinase; arylamine N-acetyltransferases; (Plummer et al. 6015; Chaudhary 1-5), amyloid protein; activating protein-1. Androgen receptor; lymphoma cell protein 2; Ca2+PK, Ca2+-dependent protein kinase; binding protein; aminopeptidase all perform important functions (Plummer et al. 6015; Chaudhary 1-5 & Druker et al. 1035). Cystic fibrosis transmembrane conductance regulator; cyclooxygenase; protamine kinase; connective tissue growth factor; chemokine work in collaboration with receptor cells to aid utilization of herb (Korutla 770; Shishodia et al. 250 & Anand 134).
Molecular Aspects of Curcumin
According to Shishodia et al. 250, Various studies have indicated that curcumin modulates multiple targets. Some of the deal with growth factors, growth factor receptors, transcription factors, cytokines, enzymes Chen et al. 278-287, and the genetic makeup is controlling apoptosis. In cytokines and growth, lots of growth factors have been associated with the growth and promotion of tumors. Curcumin has been shown to lower the expression of numerous cytokines such as TNF, IL-6, IL-8, IL-12 (Chen et al. 278-287), as well as fibroblast growth factor-2.6 Receptors. Curcumin has been shown to downregulate both epithelial growth factor receptors. It also modulates androgen receptors.
Conclusion
Considering the contributions of curcumin to the fight against cancer with no known side effects, it will be a prudent idea to promote its use. This study offers a scientific platform to support this naturally occurring substance. It is hoped that once the data is collected and analyzed, more experimental data will be unearthed to promote it further.
Work Cited
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