Introduction: Cancer begins and spreads with the uncontrolled growth of cells in the body. This is a simpler definition of cancer
Be that: The abnormal growth of the body's cells begins by ignoring the normal rules of cell division of the body. Normal cells of the body are constantly on
Now they are producing signals that indicate whether this cell should divide or differentiate into another cell or its lifespan.
The cell is finished or not. But do cancer cells pay attention to these signals or do they produce them themselves?
Which causes abnormal growth of cells and causes cancer] 1
Long unencrypted A (lncRNAs) play an important role in tumor progression and are suddenly expressed in various cancers.
To be. However, the functional role of lncRNAs in breast cancer remains largely unknown [16].
Traditional methods of treating cancer, including radiotherapy, chemotherapy, and immunotherapy, all have their own limitations. An approach
New is bacterial therapy, either used alone, or in combination with conventional methods, has a positive effect on tumor regression and inhibition
It has metastasized. Targeted treatment of the tumor with the help of bacteria, which is used as a therapeutic / gene / drug vehicle, is very promising.
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In the treatment of tumors. It was found that the use of bacteria or in combination with conventional methods in some experimental models of cancer (regression)
Tumor and increase survival (effective] 17.]
Research on bacterial toxins is closely linked to the birth of immunology. Our understanding of the interaction of bacterial protein toxin with immune cells
To decipher the immunological pathology, to develop preventive and curative treatments for infections, and to suggest anti-cancer immunotherapy methods.
Helps. More recently, immunotoxins have been used to kill cancer cells targeted by specific antibodies or cytokines.
Have been designed and used [18]
Tumor microenvironment (TME) has the ability to act as a stimulant for breast cancer progression and metastasis. TME of stromal cells
Extracellular matrix and soluble c-tokens, chemokines and extracellular vesicles and nanoparticles are formed that actively affect
Affects cell activity. Extracellular vesicles contain large exosomes, microcycles, and oncosomes that develop during tumor progression.
They regulate basic processes by interacting directly with target cells. Breast cancer exosomes by angiogenesis, invasion of the system
Strengthen immunity and resistance to chemotherapy and metastasis to the tumor. Exosomes in almost all physiological fluids, including plasma,
Urine, saliva and breast milk are found and provide a valuable source for the development of non-invasive cancer biomarkers [19]. ]
Chronic inflammation is associated with the presence of foam cells in many infectious and metabolic diseases and some cancers. These cells form when
The intracellular fat content of macrophages exceeds their capacity to maintain homeostasis. Decreased vital macrophage safety functions
Finds. Patterns of foam cell formation are caused by atherosclerosis. Recent studies show that the mechanisms of foam cell biogenesis in
The length of tuberculosis differs from that of those working during atherogenesis [20].
Worldwide, breast cancer is one of the leading causes of premature death and death in women. In the United States, breast cancer is more prevalent than anywhere else
Other than lung cancer, it can lead to cancer death in women. Various factors for breast cancer are found through epidemiological studies
It includes race, ethnicity, family history of cancer, genetic characteristics as well as alcohol use, physical inactivity,
Exogenous hormones have been shown in some women. Reproductive factors such as younger age at pregnancy and older age at first complete pregnancy can
Through long-term effects on the level of sex hormones or other biological mechanisms affect the risk of breast cancer. Cancer
Triple negative breasts may have a specific cause. Genetic types and mutations in genes that make proteins that are in the pathways
DNA repair is involved and a homologous combination of double-stranded interrupts (APEX1 DNA, BRCA1, BRCA2, XRCC2, XRCC3
ATM, CHEK2, PALB2, RAD51, XPD,) is involved in some cases of breast cancer [21.]
Tumor-associated macrophages (TAMs) aerobic glycolysis and apoptotic resistance of breast cancer cells through extracellular vesicle transfer
EV) increase the translocation of a specific myeloid lncRNA, 1α-HIF RNA stabilizer without longitudinal encoding (HISLA). In terms of
Mechanically, HISLA blocks the interaction of PHD2 and 1α-HIF to inhibit hydroxylation and degradation of 1α-HIF. Electate released from
Glycolytic tumor cells rearrange HISLA in macrophages and form a feeding link between TAM and tumor cells
gives . Blocking HISLA transmitted by EV inhibits glycolysis and chemical resistance of breast cancer in vivo. From
Clinically, HISLA expression in TAMs is associated with glycolysis, poor chemotherapy response and shorter survival in breast cancer patients
Is. Our study examines the potential of lncRNAs as signal transducers between immune cells and tumors via electrical transmitters
Transmitted to enhance cancerous aerobic glycolysis [22]
The role of some bacterial toxins in inhibiting cancer cell proliferation has been identified. The most well-known function of the toxin
Botulinum has its effects on cell integrity and cell skeleton. In 2013, Bandala and colleagues discovered the effect of Botulinum toxin
Showed proliferation and apoptosis of T47D cancer cells. They indicated that Botulinum treatment may be an alternative
Be considered as a common treatment for breast cancer. However the molecular pathways induced by Botulinum which
Its cytotoxic activity is effective. Not well identified.
Delivery
Efficacy of miRNA mimics and / or antagonists to tumor cells is a major challenge in treatment-based
miRNA is cancer. Current Approach to Delivery of Gene-Based Therapies and RNAi-Based Therapies
It is a viral or non-viral carrier system. Methods that directly utilize viral vectors cause transmission
They become efficient genes, although sometimes they have inefficiencies. Limitations on the use of the viral approach to gene transfer
And recombinogenic 2, cytopathic 3 associated with defects in tumor targeting and residual viral elements that can be immunogenic
4
Be) 24.)
The use of non-viral vectors in gene transfer may not have some of the problems associated with non-viral vectors. In this
Significant advances in the development of pharmaceutical formulations and non-viral systems for in vivo in gene therapy
It has been found that in particular, gene transfer systems due to cationic liposomes can be mentioned. Cationic liposomes
They consist of two positively charged lipid layers and can be exposed to naked DNA, which is negatively charged, by simple lipid mixing.
And DNA complexes, so that the resulting complex (lipoplex)
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(Has an overall positive charge) 20.) Lipoplex easily
It is attached to the cell and is absorbed by the cell with high transfection efficiency. The properties of cationic liposomes that make them for
DNA delivery is attractive and adaptable. The following can be mentioned:
• Ease of preparation;
• Ability to form complexes with large amounts of DNA;
• Practical adaptability to any type and size of DNA or RNA;
• Ability to transfect different types of cells, including indivisible cells;
• Lack of immunogenicity
Or inactivity biohazard 6
7
.) 25, 24 (
Currently, several clinical trials are underway using cationic liposomes to deliver genes.
Also liposomes for delivery of chemotherapeutic agents such as Doxorubicin for breast cancer chemotherapy already
8 There is an important drawback to the use of cationic liposomes is that they lack tumor specificity.
Are and in proportion
They have low transfection efficiency compared to viral vectors. However, the tumor specificity of lipoplexes can be carried by ligand transport
9, which is detected by the cell surface receptor, increases dramatically. Receptor-mediated endocytosis
Cell entry path
Highly efficient in eukaryotic cells. Ligand placement on lipoplex prevents DNA from entering cells through initial binding
The ligand facilitates the cell surface receptor and subsequent entry of the attached lipoplex into the cell. Upon entering the cell, DNA
It goes out of the endocytosis pathway to be expressed in the cell nucleus (24, 25).
In order to effectively deliver DNA to the cell, tumor-specific DNA-nanoparticle lipoplex systems with ligand targeting capabilities and
Developed for cancer gene therapy) US patent number: 6749863 and European patent number: 10 self-assembly
Or scFv against transferrin receptor 11 1154756 EP [) 26, 27. ] These nano-transport systems, transferrin
In the case of cancers 12
[22, 23] using transferrin are overexpressed as 13 humans, as a targeted ligand for the tumor
A target ligand, Xu et al. [26] Self-assembled nanocarriers in 50 to 90 nm sizes with a
They were new and obtained like a viral particle with 14 densities and a desirable surface load. These nanocarriers have nanostructures
They are dense centers surrounded by a membrane covered with transferrin molecules on the surface. This nano-carrier system is efficient and
To cancer cells in 15 promising properties for targeted delivery of antisense genes and oligonucleotides
Shown in vivo environment [28.]
One of the most controversial topics in the field of nanobiotechnology is the introduction of an efficient, safe and secure carrier for drug and gene transfer to tissues.
Is the goal. So far, various nanocarriers of polymer, fat, protein, polysaccharide, metal, etc. have been introduced for the purpose that each
Which, either due to toxicity or low absorption in human cells or rapid clearance by the immune system, is very efficient in transmission
They lacked drugs and genes, and this effort continues among researchers to introduce more efficient systems. Nanoparticles used
To transmit drugs and genes, they must be stable, biocompatible, biodegradable, non-inflammatory, non-toxic. Also the ability to escape from
Have a reticuloendothelial system so that they can circulate longer in the blood. One of the important features of targeting ability
Is a specific and successful target cell or tissue that, if a carrier has such a property, has a high score and rank among carriers.
Will no longer enjoy. Virus-derived structures have long been considered one of the most efficient drug and gene carriers.
However, due to their immunogenicity and the potential risk to them, the importance of these natural nanocarriers for systems
Transfer was reduced and researchers turned to synthetic structures such as metal nanoparticles and polymers. Gradually the toxicity of these structures
It also prevented the development of these systems and their therapeutic use (29, 30).
Today, researchers believe that this goal can be achieved only by modeling the biological structures in the human body. One
These biological nanostructures that play their role efficiently in the transfer of macromolecules and nucleic acids between cells
Are exosomes. These natural nanowicels in the human body are responsible for the transport of proteins, RNAs, and in some cases DNA.
It is responsible from one cell to another. They are completely stable, as reported after 6 months of purification and maintenance
They have not undergone any change in their structure. Because it is found in all body fluids, it is completely non-immunogenic
They will go and will not carry the risk of pathogenicity that viral vectors had. Most important of all exosomes because through fusion
And membrane integration, transport their cargo directly to the cell cytosol, thus preventing them from invading and clearing the environment.
They retain cells and do not engage in the process of escaping the endosomal structure (31). Katakowski et al.
Exosomes secreted by mesenchymal stem cells were used to load 146b-Mir.
This method of miRNA treatment using xerosomes can effectively prevent tumor growth. In addition to siRNAs and
10 Self-assembled
11 Transfer
12 Transfer receptor
13 Tumor-targeting ligand
14 Nanostructure
15 Anti-sense oligonucleotide
10
miRNAs, mRNAs can also be transported by exosomes as a commodity. (32) Recently secreted in HEK-293;
Inhibited DNA synthesis in mice and improved tumor cell apoptosis. 33) Exosomes can also be types
Load other drugs as well. Example In 2013, Zhuang and colleagues found that curcumin in lymphocyte exosomes
Loaded mice can be successfully transferred to brain tissue and improve the apoptosis of microglia cells in the brain. results
Their work suggests that this strategy may provide a new non-invasive and therapeutic approach to treating inflammatory brain diseases.
34) There are also other studies on the effect of genetic content of exosomes in the treatment of cancer, all of which confirm its effectiveness.
Exosomes are involved in gene transfer (35, 36).
Methods: 1- Separation of extracellular microvesicles from cells derived from breast carcinoma: based on centrifuge protocol
Differential and Quick Exo extraction extraction separation kit is performed by Biosciences System Company.
2 - Confirmation of the accuracy of extraterrestrial microuricles isolated in terms of size and morphology by electron microscopy:
For microscopic observation, a small volume of purified exosome is fixed with 2.5% glutaraldehyde and washed with PBS.
The sample is then dewatered with ethanol on a dry glass surface and covered with a thin layer of gold. Size and morphology
Exosomes are evaluated by electron microscopy.
3- Evaluation of HOTAIR oncogene expression in tumor microvesicles: RNA extraction: In order to extract RNA in this study from
Trizol (Invitrogen) was used. Trizol solution formed RNA complexes with 16 reactant molecules.
It is guanidinium and water, but prevents the hydrophilic binding of RNA to DNA and proteins. Then DNA and proteins
They are separated from the aqueous phase while the RNA molecules remain in this part.
4-Synthesis of cDNA from samples using TAKARA kit and study of HOTAIR expression by time-Real PCR
5- Preparation of 231-MB-MDA breast cancer grade from Iran Sastor Institute
6 - Evaluation of cytotoxicity of bacterial enzyme toxin A Cytolysin (Sigma Company) on the breast cancer strain
231-MB-MDA by assay MTT
It will be 17 micrograms per fruit to calculate the optimal dose and IC50. MTT test to evaluate cell proliferation
Then
It is treated with an external agent. The basis of this technique is based on reduction of MTT yellow substrate to purple sediment
By the enzyme mitochondrial dehydrogenase succinate in living cells. Dye intensity produced after dissolving 18 formazan
Formazan deposition in organic conditions such as dimethyl sulfoxide can be measured by spectrophotometry
And is directly related to the number of living cells and cell proliferation.
7- Loading of toxin (Sigma Company) into extracellular vesicles by sonication / incubation method
8- Treatment of breast cancer cells 231-MB-MDA in the following groups: 1- Cell group treated with foreign vesicles
Cell 2 - Toxin-treated cell group 3) Cell group with extracellular vesicles containing toxin 4) Control cell group
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16 Reagent
17 Cell proliferation
18 Formazan
14
10-Study of expression of transcripts of asostosis-related genes (2-BCL and BAX) by time-Real PCR (Housekepping gene:
GAPDH:) Time-Real PCR technique based on quantitative measurement of amplified product during the exponential phase of PCR reaction from
The way to measure the amount of light emitted is fluorescence. As the meaning of the word implies, the concept of time-real PCR is observed
Moment by moment is the process of reproduction. In this diagnostic system, a fluorescent material that can be measured is used
it is possible. During the reaction, this fluorescent dye emits fluorescent light in proportion to the amount of products produced from each cycle and the amount
Its fluorescence emission is detected and recorded by the device indicator. Time-Real PCR for target and reference genes
Is determined. In the following, 19 examples are performed. At the end of the multiplication and based on the drawn diagram, the threshold cycle
After calculating the difference between the mean CT of the reference gene and the mean CT of the target gene for both control and test samples, the ΔCT index in two
A sample of control and testing is obtained. Also, from the difference between the two ΔCTs, an index called ΔΔCT is calculated.
11 - Focytometric evaluation of treated cells in the above groups
12-Data analysis
Results: Exosomes are a good tool because they only grow and are secreted into cancer cells, so that we can send the title of therapeutic drugs to cancer cells with the help of these exosomes and make treatment easier and make it possible for It allows us not to damage the rest of the Bern cells
Conclusion: Our research needs further investigation and testing. To reach the effectiveness and treatment of NP