Inhibition of target crtac1 in lung adenocarcinoma and its application
1. The technology of CRTAC1 and lung adenocarcinoma is applied in the direction of microbial measurement/testing, medical preparations containing active ingredients, instruments, etc., to achieve the effect of auxiliary diagnosis of lung adenocarcinoma
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Embodiment 1
[0064] Example 1. The expression of CRTAC1 is consistently down-regulated in lung adenocarcinoma tissues and cell lines
[0065] 1) Use GraphPad Prism software to analyze the expression of CRTAC1 mRNA in 58 pairs of lung adenocarcinoma patients in the TCGA database by t-test results. Pfigure 1 A).
[0066] 2) Tissue samples
[0067] The clinical samples of lung adenocarcinoma used in the present invention were provided by a cooperative hospital, and the tissue was diagnosed as lung adenocarcinoma by a pathologist. When the specimen was collected, a position about 3 cm around the lung adenocarcinoma tissue was selected as the paracancerous tissue control, and a part of the tissue was fixed with 4% PFA, and the rest was frozen at -80 degrees for later use. All patients have detailed information such as name and pathological information. The experimental operation was approved by ethical review.
[0068] 3) Tissue / cell total RNA extraction
[0069] 1. Take out the collected l...
Embodiment 2
[0098] Kaplan-Meier survival analysis was performed on the survival information of CRTAC1 in 401 patients with lung adenocarcinoma in the TCGA database using GraphPad Prism software. The results of the overall survival rate of lung adenocarcinoma samples showed P figure 2 A). That is, the expression of CRTAC1 was significantly positively correlated with the survival time of patients with lung adenocarcinoma ( figure 2 A).
Embodiment 3
[0099] Example 3. Overexpression of CRTAC1 significantly inhibits the proliferation ability of lung adenocarcinoma cells
[0100] 1) Select H1299 and A549 cells as research objects, infect GFP-CRTAC1 plasmid and GFP empty plasmid virus supernatant in H1299 and A549 cells, and use Puromycin drug screening to establish stable cell lines H1299 (CRTAC1), A549 (CRTAC1) and The control cell lines H1299 (Vector) and A549 (Vector). Western Blot was used to detect the overexpression effect, and β-Actin was used as an internal reference, such as image 3 A and 3D are shown.
[0101] 2) The soft agar colony formation experiment was used. At room temperature, a medium was prepared in a clean bench, and 12.6ml (1.8ml×7) was preheated at 45-46°C for more than 10min. Microwave heating and boiling agar for 3 times, after cooling to 45-46°C, take 8.4ml (1.2ml×7) of agar and mix quickly in a medium preheating tube, absorb 3ml / well and spread the next layer of agar glue, and let it solidify natu...
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