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miR-129 and application of target genes of miR-129 to detection of lung adenocarcinoma
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A lung adenocarcinoma and target gene technology, applied in the application field of miR-129 and its target gene in the detection of lung adenocarcinoma, can solve the problem of low survival rate of NSCLC
Active Publication Date: 2019-01-08
WEST CHINA HOSPITAL SICHUAN UNIV
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[0004] Lung adenocarcinoma is a type of non-small celllung cancer (NSCLC) with a relatively high incidence. At present, the treatment of NSCLC mainly adopts surgery, combined with radiotherapy, chemotherapy, biological immunotherapy and other treatment methods. Although the treatment methods are constantly improving, But the five-year survival rate of NSCLC is still very low, only about 15%
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[0056] The 6 cases of lung adenocarcinoma tissues and adjacent tissues were all obtained from hospital surgically resected specimens. All specimens were placed in liquid nitrogen tanks within 10 minutes of isolation, and then transferred to a -80°C refrigerator for storage.
[0059] 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.
[0060] 2) Add 1 / 10 volume of Homogenate Additive to the above homogenized tissue samples, and place on ice for 10 minutes.
[0061] 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.
[0062] 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.
[0063] 5) Add 700 μl of miRNA eluent 1 to the spin co...
[0073] 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.
[0074] According to the statistical analysis of the data provided by the sequencing company, FDR1, and the difference between the average counts of the two groups was greater than 100. For the differentially expressed miRNAs, miRNAs with significant differential expression in the filter were artificially selected, and the molecular marker miR-129, which is very well correlated with lung adenocarcinoma, entered our research scope, and the expression level of miR-129 in lung adenocarcinoma tissues was higher than that of Paracancer.
[0075] Identifying miRNA target genes is a very important step in studying miRNA function in specific tissues and cells. Using algorithms including RNA22, miRanda, miRDB, miRWalk, PICTAR2 and Targetscan to predict t...
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Abstract
The invention relates to miR-129 and an application of target genes of the miR-129 to detection of lungadenocarcinoma. A high-flux sequencing method is used for seeking genes being obvious in difference expression in the lungadenocarcinoma and tissues close to the lungadenocarcinoma. In combination of bioinformatics analysis and literature investigation and research, miR-129 and the target genes MFAP3L and RPH3A of the miR-129 are selected from the candidate miRNA and are subjected to RT-PCR verification and databaseverification. Results indicate that the miR-129 and the target genes thereof have good relevance to the lung adenocarcinoma. The invention provides new important target points for gene diagnosis and treatment of the lung adenocarcinoma for clinical treatment.
Description
technical field [0001] The invention relates to the field of molecular diagnosis, in particular to the application of miR-129 and its target gene in detecting lung adenocarcinoma. Background technique [0002] miRNA is a non-coding small RNA consisting of about 19 to 25 nucleotides, which plays an important regulatory role in animals and plants by inhibiting the translation or degradation of its target mRNA. miRNA is one of the important components of many generegulatory molecules in multicellular organisms, and it is likely to affect the expression output of many protein-coding genes. miRNA can be divided into oncogenic miRNA and tumor suppressor miRNA according to its regulatory role in the occurrence and development of tumors, which regulate the proliferation, invasion, apoptosis and angiogenesis of cancer cells. [0003] miR-129 is highly expressed in many common tumors, such as gastric cancer and lung cancer, or low expressed, such as breast cancer and esophageal squa...
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