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Application of mettl14 gene as a biomarker in the preparation of prognostic detection preparations for lung adenocarcinoma

A technology of lung adenocarcinoma and detection kit, which is applied in the field of tumor molecular biology, can solve the problems of inability to adapt to the prognosis judgment of lung adenocarcinoma patients, no specific indicators, and no reference standard for the prognosis judgment of lung adenocarcinoma patients, etc. Profound clinical significance and promotion effect

Active Publication Date: 2021-11-05
XIANGYA HOSPITAL CENT SOUTH UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there is no reference standard for the prognosis of patients with lung adenocarcinoma, and there is no specific index, which is far from meeting the needs of the prognosis of patients with lung adenocarcinoma

Method used

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  • Application of mettl14 gene as a biomarker in the preparation of prognostic detection preparations for lung adenocarcinoma
  • Application of mettl14 gene as a biomarker in the preparation of prognostic detection preparations for lung adenocarcinoma

Examples

Experimental program
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Embodiment 1

[0013] Example 1 Preparation of METTL14 gene for lung adenocarcinoma prognosis detection kit (50 reactions)

[0014] 1. RNA stabilization solution 50ml

[0015] 2. Isopropanol 100ml

[0016] 3. Chloroform 100ml

[0017] 4. Trizol 50ml

[0018] 5. Enzyme-free water 10ml

[0019] 6. 1μM random reverse transcription primer 50ul

[0020] 7. 5× reverse transcription buffer 200ml

[0021] 8. 10mM base triphosphate deoxyribonucleotides 100ul

[0022] 9. 40U / μl RNase inhibitor 500ul

[0023] 10. 200U / μl MMLV reverse transcriptase 50ul

[0024] 11. Premix Ex Taq 50ul

[0025] 12. 10μM METTL14 gene real-time fluorescent quantitative PCR specific primers:

[0026] Forward primer 5'-CTGAAAGTGCCGACAGCATTGG-3' (SEQ ID NO.2),

[0027] Reverse primer 5'-CTCTCCTTCATCCAGATACTTACG-3' (SEQ ID NO.3),

[0028] 13. 10μM internal reference GAPDH specific primers for real-time fluorescence quantitative PCR:

[0029] The forward primer is 5'-GGTCGTATTGGGCGCCTGGTCACCAG-3' (SEQ ID NO.4),

[...

Embodiment 2

[0031] Example 2 Detection of METTL14 gene in lung adenocarcinoma tissue

[0032] 1. Preservation of lung adenocarcinoma tissue: collect the lung adenocarcinoma tissue to be tested and store it in a cryopreservation tube filled with RNA stabilization solution, and put it in a -80°C refrigerator for later use.

[0033] 2. Extraction of RNA in tissues: Take an appropriate amount of specimen, add liquid nitrogen to the mortar after baking at 180°C for 6-8 hours, grind the specimen, grind to powder, add 1ml Trizol mortar specimen to the mortar, and grind into After the liquid is in the centrifuge tube, add 200μl / ml chloroform to the centrifuge tube, shake it by hand for 15-30s, place it on ice for 5min, and centrifuge at 12000g at 4°C for 15min; carefully take the upper aqueous phase into a new centrifuge tube, add Mix with 0.5ml of pre-cooled isopropanol, put it in a refrigerator at -20°C for 20min, centrifuge at 12000g at 4°C for 10min; discard the supernatant, add 1-2ml of etha...

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Abstract

The invention discloses the application of the METTL14 gene as a biomarker in the preparation of a lung adenocarcinoma prognosis detection preparation. The expression level of METTL14 gene in patients with lung adenocarcinoma can be detected, which provides a strong molecular biological basis for predicting the prognosis of patients with lung adenocarcinoma, and has far-reaching clinical significance and promotion.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and specifically relates to the application of the METTL14 gene as a biomarker in the preparation of lung adenocarcinoma prognosis detection reagents. Background technique [0002] Lung cancer is currently the malignant tumor with the highest mortality rate. The incidence of lung cancer in my country has been increasing year by year, and it has become the malignant tumor with the highest morbidity and mortality. Non-small cell lung cancer accounts for 80% to 85% of all lung cancers. The median survival time of advanced non-small cell lung cancer is about 10 months. So far, there has been no breakthrough in its treatment and prognosis. Non-small cell lung cancer is mainly divided into lung adenocarcinoma and lung squamous cell carcinoma according to different histological types, among which lung adenocarcinoma is the most common type of non-small cell lung cancer. Although the advancement o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/118C12Q2600/158
Inventor 陶永光王敏毛超刘双
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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