Composition related to lung adenocarcinoma metastasis and application thereof
A technology of lung adenocarcinoma and composition, which is applied in the composition and its application fields related to the metastasis of lung adenocarcinoma, and can solve the problems affecting the expression of target genes
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Embodiment 1
[0065] The collection of embodiment 1 sample
[0066] The tumor tissues of 10 cases of lung adenocarcinoma were all obtained from surgically resected specimens of Beijing Friendship Hospital from January 2009 to December 2009. All specimens were placed in a liquid nitrogen tank within 10 minutes of isolation, and then transferred to a -80°C refrigerator Storage, after 3 years of follow-up, no metastasis occurred in 3 patients, and metastasis occurred in 7 patients.
Embodiment 2
[0067] Example 2 total RNA extraction
[0068] 1 extraction method
[0069] 1) Take 80 mg tissue block, add 800 μl Lysis / Binding buffer, and use a homogenizer to homogenize the tissue block. During the homogenization process, the samples should be placed on ice to keep the cold state.
[0070] 2) Add 1 / 10 volume of Homogenate Additive to the above homogenized tissue samples, and place on ice for 10 minutes.
[0071] 3) Add an equal volume of water-saturated phenol to the Lysis / Binding buffer, shake for 45 seconds, and centrifuge at 10,000×g for 5 minutes at room temperature.
[0072]4) Carefully remove the supernatant into a new test tube, add 1.25 times the volume of absolute ethanol, mix well, transfer to a purification column, centrifuge at 10,000×g for 15 s, and discard the liquid in the collection tube. Since the maximum volume of the column is only 700 μl, repeat this step until all the supernatant is filtered.
[0073] 5) Add 700 μl of miRNA eluent 1 to the spin col...
Embodiment 3
[0082] Example 3 Sequencing and Data Analysis
[0083] The sequencing company carried out the establishment of the sequencing library and on-machine sequencing, and the sequencer used was the HiSeq2000 sequencer of Illumina Company.
[0084] According to the data analysis results provided by the company: the metastatic group (7 cases) and the non-metastatic group (3 cases) were statistically analyzed, the P value was less than 0.05, and the difference between the metastatic group and the non-metastatic group was at least more than 2 times of differential expression miRNA, mir-193a, mir-3942, mir-767, mir-193b, mir-550a-1 with significantly up-regulated expression in the artificial selection and filtering of differentially expressed miRNAs entered our research scope.
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