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BRM270 when combined with the use of miRNA-128, which acts as a tumor suppressor downregulated in lung cancer would be a great way to combat chemoresistance of NSCLC [19]. Instead of using the virus we could use this compound combined with miRNA-128 to combat the resistance developing of gefitinib. Increases sensitivity of TKI. BRM270 has a low toxicity and targets the NF-kB signaling pathway, induces cell cycle arrest and apoptosis. This is combined with the downregulation of marker CD133. Since we want to suppress Vimentin in our initial method using BRM270 will be another way to suppress Vimentin and induce E-cadherin which is good because then this will mean we are progressing to inhibiting metastatic progression.

BRM270 is made up of 7 distinct types of plants known as Saururus chinensis,, Aloe vera, Arnebia euchroma,Citrus unshiu Markovich, Portulaca oleracea, Scutellaria baicalensis, and P.vulgaris var. Lilacina. Studies have shown that this substance plays a role in inhibiting tumor cell proliferation [19]. Some of these extract plants are known to be involved in anti-inflammatory activity like P.vulgaris var. Lilacina [20] BRM270 has been illustrated to suppress formation of tumors and does this by negatively regulating nuclear factor-kB signaling in drug resistant cancer stem cells. A study was conducted in which lung adenocarcinoma cells migration and invasion to other areas as well as chemoresistant derivatives was inhibited by treating it with BRM270.

Therefore having a lower recruitment of neutrophils in the lung could result in a decrease of lung metastasis with TLR3 already inhibited. This then suggest that TLR3 is also involved in promoting pre-metastatic niche creation by recruiting neutrophils to the lung [7]. When TLR3 is present in host stromal cells than chemokine production is high and this then means that neutrophils have an easy way to mobilize and be recruited which we already know are known to contain Bv8 and this will then mean that transportation of tumor cells and facilitation is increased as well [7]

Neutrophils are involved in cellular mediating in damaging microorganisms. It damages cells and tissues of the host. It also contains properties involved in healing wounds and they make key contributions to recruiting and then activating and programming dendritic cells and macrophages. Then the adaptive immune system does its job and regulates the rate of neutrophils being produced in bone marrows. [10] Neutrophils have been studied to be involved in facilitating lung cancer metastasis. The main bone marrow derived cells  found in the metastatic niche are known as cKIT+ and vascular endothelial growth factor receptor 1 (VEGFR1+ ) myeloid cells. Bv8 is also present in pro-metastatic neutrophils.

To see whether TLR3 is involved in pre-metastatic niche formation, the expression of Bv8 was studied since it is involved in promoting primary tumor cell migration and invasion to the potential metastatic site [7]. Studies concluded that TLR3 inhibited had a decrease in the expression of Bv8 which means that transportation of the tumor cells was decreased as well. It also was illustrated that a protein known as fibronectin was downregulated which means that this protein involved in binding BMDCs and tumor cells in pre-metastatic niche was inhibited [7] . It has been known that bone marrow cells bring tumor cells to different sites to form metastatic sites.

DNA extraction is a process used in many laboratory settings. It involves the purification of DNA from a sample using a combination of chemical and physical methods. The purpose of DNA extraction is to obtain DNA in a relatively purified form. Once the purifed form is obtained it can then be analyzed using PCR or sequencing the DNA. In a DNA extraction procedure there are essential components. Theses componenets include maximizing DNA recovery, removing inhibitors, removing or inhibiting nucleases, maximizing the quality of DNA, and double strand vs. single strand (RFLP or PCR). There are a variety of commonly used DNA extraction procedures. Any method one decides to use may be sample dependent, technique dependent, cost depedndent, or analyst preference. 

The solution was permitted to dry for 5 minutes and the product was transferred via pipet into a tared new vial. The mass was recorded and the percent recovery was calculated (0.389 g, 18.87%). The chemical tests known as bromine in dichloromethane and potassium permanganate tests for alkene were performed. Cyclohexene product (~0.25 mL) was added to one test tube and cyclohexane (~0.25 mL) was added to another test tube. 3% solution of bromine in dichloromethane (3 drops) was added into each tube while swirling.

To perform an acid-catalyzed dehydration of cyclohexanol with phosphoric acid to form cyclohexene and water/hydronium. The presence of an alkene will be identified through reactions with bromine in dichloromethane and potassium permanganate. For the potassium permanganate test, cyclohexene product (0.3 mL) was added into a new test tube and an additional cyclohexane (0.3 mL) was added to another tube. 1% potassium permanganate and 10% sulfuric acid (~2 drops) was added to each test tube while swirling.  Color changes were recorded. The gas chromatography and infrared analysis readings were received to indicate and illustrate the degree of purity of the product.

Most basic of all procedures and involves purification of DNA. An important step within organic extraction is the removal of proteins. Can be done by extracting aqeous solutions of nucleic acids with phenol or chloroform. There are pros and cons when dealing with organic extraction. The pros are it yields relatively pure, high molecular weight DNA. In addition, DNA is double stranded. This means it is good for RFLP and PCR. The cons are it is time consuming and there is the increase risk of contamination. It also involves use of hazardous chemicals. 

The purpose of DNA extraction is to obtain DNA in a relatively purified form. This can then be used to analyze using PCR or sequencing the DNA obtained. In a DNA extraction procedure there are essential components which are to maximize DNA recovery, remove inhibitors, remove or inhibit nucleases, maximize the quality of DNA, and double strand vs. single strand (RFLP or PCR). There are a variety of most commonly used DNA extraction procedures. The method utilized may be sample dependent, technique dependent, cost depedndent, or analyst preference.