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Lung adenocarcinoma associated miRNAs, composition and application of miRNA and composition
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A technology of lung adenocarcinoma and composition, applied in the field of molecular biology
Inactive Publication Date: 2017-08-18
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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In recent years, with the continuous development of precision medicine, the individualized treatment of lung cancer requires the precise classification of lung cancer at the molecular level. Although there have been many reports disclosing some molecular markers for the diagnosis of lung cancer, they still cannot meet the needs and need more. Molecular markers help molecular typing, targeted therapy and prognosis of lung cancer
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Embodiment 1
[0051] Example 1 Collection of samples
[0052] The 4 cases of lung adenocarcinoma tissues and adjacent tissues all came from specimens surgically removed from Beijing Friendship Hospital from January 2015 to July 2016. All specimens were placed in a liquid nitrogen tank within 10 minutes of being isolated, and then transferred to -80 Store in refrigerator at ℃.
[0055] 1) Take 80mg tissue block, add 800μl Lysis / Binding buffer solution, and homogenize the tissue block with a homogenizer. During the homogenization process, the sample should be kept on ice to keep it cool.
[0056] 2) Add 1 / 10 volume of Homogenate Additive to the homogenized tissue sample, and place it on ice for 10 minutes.
[0057] 3) Add the same volume of water-saturated phenol as Lysis / Binding buffer, shake for 45s, centrifuge at 10,000×g for 5min at room temperature.
[0058] 4) Carefully take out the supernatant into a new test tube, add 1.25 times the volume of absolute ethanol, and after mixing, transfer it to the purification column, centrifuge at 10,000×g for 15 seconds, and discard the liquid in the collection tube. Since the maximum volume of the column is only 700μl, repeat this step until all the supernatant is filtered.
[0059] 5) Add 700μl miRNA eluate 1 to the centrifuge column, centrifuge at ...
[0069] The sequencing library was established and sequenced on the computer by the sequencing company, and the sequencer used was the HiSeq2000 sequencer of Illumina company.
[0070] Statistical analysis based on the data provided by the sequencing company, fdr 1. The difference between the two count averages is greater than 100. For differentially expressed miRNAs, we artificially select and filter the differentially expressed miRNAs. Several molecular markers related to lung adenocarcinoma that have never been reported before have entered the scope of our research. Among them, miR-6087 and miR-7976 are in cancer tissues. The expression of miR-7705 and miR-6744-5p in cancer tissues is higher than that in adjacent tissues.
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Abstract
The invention relates to lungadenocarcinoma associated miRNAs, composition and an application of the miRNAs and the composition, in particular to an application of miR-6087, miR-7976, miR-7705 and miR-6744-5p and composition of miR-6087, miR-7976, miR-7705 and miR-6744-5p to preparation of diagnostic preparations for lungadenocarcinoma. Samples of cancerous tissue and adjacent tissue are subjected to transcriptome sequencing and analysis is performed with a bioinformatic method, a plurality of unreported lungadenocarcinoma associated molecular markers are discovered, molecular verification and ROC (receiver operating characteristic) curve analysis prove that the newly discovered lung adenocarcinoma associated miRNAs can become potential molecular markers for lung adenocarcinoma, and basis is provided for clinical diagnosis.
Description
Technical field [0001] The present invention relates to the field of molecular biology, specifically to miRNAs, compositions and applications related to lung adenocarcinoma, and more specifically to miR-6087, miR-7976, miR-7705, miR-6744-5p and their combinations Application in the preparation of lung adenocarcinoma diagnostic preparations. Background technique [0002] Lungcancer is one of the malignant tumors with a high mortality rate. It mainly includes small celllung cancer and non-small celllung cancer. In clinical incidence, non-small celllung cancer is the majority, accounting for about 80%. Non-small cell lung cancer can be further subdivided into adenocarcinoma and non-small cell lung cancer. There are two major types of squamous cellcarcinoma. In recent years, with the continuous development of precision medicine, the individualized treatment of lung cancer requires precise classification of lung cancer at the molecular level. Although there have been many report...
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