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  3. Detection of lung cancer using nano biosensor based on nanohybrid graphene oxide - DNA

Detection of lung cancer using nano biosensor based on nanohybrid graphene oxide - DNA

Number of pages: 67 File Format: word File Code: 31884
Year: 2014 University Degree: Master's degree Category: Chemical - Petrochemical Engineering
Tags/Keywords: DNA - Fluorescence spectroscopy - Graphene oxide - Graphene oxide - DNA - Knockout jump - lung cancer - Nano biosensor - nano biosensor, - Nano hybrid
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  • Summary of Detection of lung cancer using nano biosensor based on nanohybrid graphene oxide - DNA

    Dissertation for Master's Degree in Organic Chemistry

    Abstract

    Today, lung cancer is considered one of the most common diseases worldwide and has the highest mortality rate among all types of cancer. Therefore, early diagnosis of this disease is of special importance. Considering that the common methods for detecting lung cancer are expensive and time-consuming, offering cheaper and faster methods has been given special attention. With the significant progress of nanotechnology in recent years and the development of various nanomaterials, activities have been carried out in this field. Recent studies show that the graphene oxide nanomaterial has a high potential in the field of designing bio-nanosensors for lung cancer detection due to its unique properties.

    In the current thesis, a bio-nanosensor based on graphene oxide-DNA nanohybrid is presented to detect the deletion mutations that cause lung cancer. In this method, mutations have been identified using FAM-labeled DNA probe and through fluorescence spectroscopy. Also, graphene oxide has been synthesized based on Hammer's method, and it has been investigated and confirmed using FT-IR, UV-Vis and TEM image spectra.

    Key words: graphene oxide, biosensor, lung cancer, DNA, deletion mutation

    -1- Lung cancer

    Cancer is a genetic disease and is caused by the uncontrollable growth and division of cells in a part of the body, which is the result of environmental factors and genetic disorders. In other words, cancer is caused by a series of successive mutations in human genes. Today, there are more than 200 types of cancer, one of the most common types of which is lung cancer.

    Lung cancer is the second most common cancer among men and women and is one of the most preventable types of cancer. In general, there are two types of lung cancer:

    1) Small cell lung cancer[1] (SCLC)

    2) Non-small cell lung cancer[2] (NSCLC)

    The way of growth and spread of both in the body and their treatment methods are different. Types of lung cancer are classified based on the appearance of the cells under the microscope. Non-small cell lung cancer (NSCLC) is also divided into three categories: 1) superficial tissue cancer [3], 2) cancer of the mucosa-infiltrating glands and lymphatic vessels [4] (glandular epithelium) and 3) large cell lung cancer [5] [1, 2]. Among people with this type of cancer, about 85-90% of cases are NSCLC and about 10-15% are SCLC.

    The most common clinical symptoms of lung cancer include persistent and chronic cough, chest pain, anorexia, weight loss, bloody sputum, shortness of breath, respiratory infections such as bronchitis, and the onset of wheezing. is, which usually does not appear in the early stages of the disease. Therefore, the death rate of this type of cancer is very high [3].

    1-1-1- Risk factors

    Tobacco use, especially smoking, is the most important risk factor for developing lung cancer [4], because almost 90% of patients with lung cancer are smokers and the risk of lung cancer is about It is 20 to 40 times more common in smokers than in non-smokers. Smoking is reported to be the main cause of lung cancer in approximately 79% of women and 90% of men, and 90% of lung cancer deaths are caused by smoking [5]. Radon gas is the second major cause of lung cancer after smoking [6], this gas is radioactive, odorless, tasteless, and colorless, and naturally does not break down. Uranium is formed in soils and rocks. According to statistics, annual exposure to this gas is involved in more than 20,000 lung cancer deaths in the United States [7]. Other things that increase the risk of this type of cancer and exist in the workplace include: polycyclic aromatic hydrocarbons, arsenic, asbestos, cadmium, beryllium, compounds containing nickel and chromium, chloromethyl ethers, etc. are [4].. Also, things like air pollution, family history of lung cancer, lung radiation therapy, improper diet, old age, hereditary and acquired genetic changes, etc. They are among the causes of lung cancer.

    1-1-2- Gene changes causing lung cancer

    During the last several years, scientists have made great progress in identifying the effect of risk factors on DNA and gene changes that lead to cancerous cells. They have determined the stages of cancer production in which several mutated genes are involved, these genetic changes disrupt the normal order of cell division and differentiation.

    Lung cancer is often the result of a series of genetic changes including the activation of proto-oncogenes and their transformation into oncogenes[6], and the inactivation of tumor suppressor genes[7] (TSGs). and is Proto-oncogenes are genes that are responsible for regulating the division and growth of cells in the normal state, and if they find a genetic mutation, they are called oncogenes, whose gene expression is very high. And tumor suppressor genes are genes that slow down cell division and determine the time of cell death. The lack of tumor suppressor genes causes cells to divide uncontrollably.

    Oncogenes that lead to lung cancer include c-myc, mutated kras (not seen in any SCLC lung cancer cases, but in 15-20% of NSCLC lung cancer cases and most cases of glandular epithelium), egfr gene overexpression. cyclin D1, BCL2 and are  Tumor suppressor genes (TSGs) involved in most cases of lung cancer include p53 (found in 90% of SCLC and 50% of NSCLC), Rb (found in 90% of SCLC and 20% of NSCLC), p16 (found in more than 50% of NSCLC and less than 1% of SCLC) and are And hTR and hTERT genes are expressed in almost all types of lung cancer as an immortal mechanism [8]. The high mortality of this type of cancer is due to the high incidence of the disease and the low chance of survival. Recently, the American Cancer Society reported more than 226,000 new cases of lung cancer and more than 160,000 deaths from it in the United States, accounting for about 27% of all cancer deaths. And according to global statistics, more than one million people die from this type of cancer every year [3, 9].

    More than 80% of lung cancer patients die in less than five years from the time of diagnosis [10], because most of them are diagnosed with the disease during the advanced stages of the disease and when its treatment is not very possible. It is of particular importance.

    1-3- Lung cancer detection methods

    Until now, in the field of medicine, various methods have been used to detect lung cancer, among which are chest x-rays [8], CT scan[9], MRI[10], bone scan[11], bronchoscopy[12], sputum test[13] and

  • Contents & References of Detection of lung cancer using nano biosensor based on nanohybrid graphene oxide - DNA

    List:

    Chapter One: Introduction and review of sources. 1-1- Lung cancer. 2

    1-1-1- risk factors 3

    1-1-2- genetic changes causing lung cancer. 4

    1-2- The importance of identifying lung cancer. 5

    1-3- Lung cancer detection methods. 6

    1-3-1- silica nanowires 7

    1-3-2- gold nanoparticles. 10

    1-3-3- carbon nanotubes. 13

    1-3-4- Quantum dots. 17

    1-4-graphene. 21

    1-5-graphene oxide. 24

    1-6-Uses of graphene oxide. 27

    1-6-1-use of graphene oxide in bioelectrochemistry. 28

    1-6-2- Medical and biological applications of graphene oxide. 29

    1-7- The purpose of the present scientific work. 38

    The second chapter: Experimental part. 39

    2-1- Materials and devices 40

    2-2- Preparation of Tris-HCl buffer 42

    2-3- Graphene oxide synthesis. 42

    2-4 Preparation of solutions to measure the fluorescence spectrum. 43

    2-4-1- Preparing the first step solution. 43

    2-4-2- Preparation of second stage solution. 43

    2-4-3- Preparation of third stage solutions. 44

    2-4-4- Preparing the solution of the fourth step. 44

    2-4-5- Preparation of solutions of the fifth stage. 44

    2-4-6- preparing the solutions of the sixth step. 45

    The third chapter: results and discussion. 46

    3-1- Preparation of graphene oxide from graphite. 47

    3-2- Examining the UV-Vis spectrum of graphene oxide. 48

    3-3- Interpreting the IR spectrum of graphene oxide. 49

    3-4- Examining the TEM image of graphene oxide. 49

    3-5- Lung cancer biomarker selection. 50

    3-6- Interpretation of emission spectra. 53

    3-6-1- Examining the DNA probe fluorescence spectrum. 53

    3-6-2- Optimization of DNA probe absorption time on GO surface. 54

    3-6-3- Optimizing the amount of GO in the presence of DNA probe. 56

    3-6-4- Examining the fluorescence spectrum of DNA-GO probe complex in the presence of target DNA (healthy DNA) 57

    3-6-5- Optimizing the hybridization time of target DNA with probe DNA in the presence of GO. 58

    3-6-6- Examining the fluorescence intensity changes of DNA-GO probe complex in the presence of different concentrations of target DNA 60

    3-6-7- Examining the fluorescence spectrum of DNA-GO probe in the presence of mDNA (mutant DNA) 62

    3-7- Identifying lung cancer. 63

    3-8- Conclusion. 65

    3-9- Suggestions. 66

    Resources 67

    Source:

     

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