ANXA3 gene or ANXA3 protein as biomarker of lung adenocarcinoma
A lung adenocarcinoma and genetic technology, applied in the determination/testing of microorganisms, DNA/RNA fragments, biological testing, etc., can solve the problem that the five-year survival rate of patients is less than 15%
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Embodiment 1
[0036] Expression of ANXA3 gene in lung adenocarcinoma
[0037] 1 Gene chip detection of gene expression
[0038] 1.1 Materials
[0039] Surgically resected cancer tissues and paracancerous tissues of 29 patients with primary lung adenocarcinoma were used as experimental samples. All patients with lung adenocarcinoma lived in Xuanwei area for more than ten years, and all cancer tissues were pathologically confirmed as lung adenocarcinoma. All patients with primary lung adenocarcinoma had not received radiotherapy and chemotherapy before operation, and the clinical data of all cases were complete.
[0040] 1.2 Extraction of tissue RNA
[0041]The RNA in each tissue was extracted by TRIZOL method, and the concentration and purity of the extracted RNA were determined by Nanodrop2000, and the OD260 / 280 was required to be between 1.8 and 2.2. Store at -80°C for later use.
[0042] 1.3 Fluorescence labeling of samples
[0043] a. Synthesis of First strand cDNA by reverse trans...
Embodiment 2
[0063] Expression of ANXA3 Protein in Lung Adenocarcinoma
[0064] 1Clinical specimen collection and lung adenocarcinoma tissue information
[0065] The collection of specimens and the preparation of case data are the same as in the first part. In this part, 21 pairs of primary lung adenocarcinoma tissue and paracancerous specimens were detected.
[0066] 2 Tissue protein extraction and concentration measurement
[0067] The RIPA lysis method was used to extract the total protein in the tissue, and the BCA protein concentration assay kit was used to determine the protein concentration, using the Biyuntian kit, catalog number: P0012S.
[0068] 3 protein denaturation
[0069] Each protein sample was adjusted according to the measured sample concentration, and an appropriate amount of 5×loadingbuffer was added to each tube, boiled in a metal bath for 5 minutes, and stored at -20°C for each use.
[0070] 4Western Blot experimental steps
[0071] Prepare SDS-PAGE gel according ...
Embodiment 3
[0076] Immunohistochemical expression of ANXA3 protein in lung adenocarcinoma
[0077] 1. Research materials
[0078] A total of 69 specimens of primary lung adenocarcinoma in the Xuanwei area were collected from the First Affiliated Hospital of Kunming Medical University from 2016 to 2017. A total of 72 cases of cancer specimens. A total of 141 cases of lung adenocarcinoma were collected. Before surgery, the patients had no history of radiotherapy and chemotherapy. The collection of all cases included the tumor part and the paracancerous area, and relatively complete clinical data were obtained. In addition, human spleen tissue was selected as the positive control photo of the ANXA3 antibody.
[0079] Among the 69 cases of primary lung adenocarcinoma in Xuanwei area, there were 43 males and 26 females, including 39 smokers and 30 non-smokers. The age ranges from 32 to 77 years old, the average age and median age are 60 and 52 years old respectively. There were 23 cases wi...
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