Target CRTAC1 for inhibiting lung adenocarcinoma and application of target CRTAC1
A 1. CRTAC1, lung adenocarcinoma technology, applied in the biological field
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Embodiment 1
[0064] Example 1. The expression of CRTAC1 is uniformly down-regulated in lung adenocarcinoma tissues and cell lines
[0065] 1) The expression of CRTAC1 mRNA in 58 pairs of lung adenocarcinoma patients in the TCGA database was analyzed by t-test using GraphPad Prism software. 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 pathological physician. During specimen collection, a position about 3 cm around the lung adenocarcinoma tissue was selected as the paracancerous tissue control. A portion of the tissue was fixed with 4% PFA, and the rest was frozen at -80°C for later use. All patients have detailed information such as name and pathological information. The experimental procedure was approved by ethical review.
[0068] 3) Tissue / cell total RNA extraction
[0069] 1. Take out the collected lung canc...
Embodiment 2
[0098] Kaplan-Meier survival analysis was performed on the survival information of CRTAC1 in 401 lung adenocarcinoma patients in the TCGA database using GraphPad Prism software. The high expression of CRTAC1 (>median; n=109) and the low expression of CRTAC1 ( figure 2 A). That is, the expression of CRTAC1 was significantly positively correlated with the survival of lung adenocarcinoma patients ( figure 2 A).
Embodiment 3
[0099] Example 3. Overexpression of CRTAC1 significantly inhibits the proliferation of lung adenocarcinoma cells
[0100] 1) Select H1299 and A549 cells as research objects, infect H1299 and A549 cells with GFP-CRTAC1 plasmid and GFP empty plasmid virus supernatant, and use Puromycin drug screening to establish stable cell lines H1299 (CRTAC1), A549 (CRTAC1) and Its control cell lines H1299 (Vector), A549 (Vector). The overexpression effect was detected by Western Blot, 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 naturally for mo...
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