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Lung adenocarcinoma miRNA marker

A lung adenocarcinoma, miRNA-2166 technology, applied in miRNA-2116 markers and their application fields, can solve problems such as abnormal expression and inducing cancer

Active Publication Date: 2015-07-15
昆明博奥三合医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The abnormal expression of miRNA directly leads to the abnormal expression of some genes related to cancer, which induces the occurrence of cancer.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Screening of miRNAs associated with metastasis of lung adenocarcinoma

[0047] 1. Sample acquisition: 30 cases of lung adenocarcinoma tissue (including 15 samples with metastasis and 15 samples without metastasis). The above samples are surgical resection specimens of patients with lung adenocarcinoma. These samples come from the Fourth Hospital of Hebei Medical University and Beijing Friendship Hospital. All the above specimens were obtained with the consent of the organizational ethics committee. The clinical data of the tissue samples include: gender, age, tumor size, pathological grade (Edmonson), metastasis or recurrence, etc.

[0048] 2. Extraction of total RNA from lung adenocarcinoma tissue

[0049] Add 1mL Trizol per 100mg of tissue, grind with liquid nitrogen to thoroughly grind the tissue pieces. Add about 1 / 5 of the volume of chloroform, mix well by inverting up and down for about 1 minute, and let stand at room temperature for 5 minutes. After...

Embodiment 2

[0058] Example 2 QPCR verification of differentially expressed miRNA-2116

[0059] 1. According to the detection results of the miRNA chip, select miRNA-2116 for large sample QPCR verification. According to the sample collection method in Example 1, 80 cases of metastatic lung adenocarcinoma tissue and 80 cases of non-metastatic lung adenocarcinoma tissue were selected.

[0060] 2. The RNA extraction process is the same as in Example 1.

[0061] 3. Reverse transcription: mix 10pg-1μg total RNA template with 2μl 10* buffer, 2μl dATP (10mM), 0.5μl polyA polymerase, 0.5μl ribonuclease (RNase) inhibitor and ribonuclease free water (RNase free water) to a final volume of 20 μl, and incubated at 37° C. for 1 h. Then add 1 μl 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, incubate at 70°C for 5 minutes and immediately incubate on ice for at least 2 minutes to break the secondary structure of RNA and primers. Finally, mix the above 20 μl reaction mixture with 4 μl 5* ...

Embodiment 3

[0065] Example 3 Expression of miRNA-2116 in lung adenocarcinoma cell lines

[0066] 1. Cell culture

[0067] Lung adenocarcinoma cell lines A549, NCI-H1650, NCI-H1299, SPC-A-1 were cultured in RPMI1640 medium and 10% fetal bovine serum, human lung epithelial cell line BEAS-2B was cultured in BEGM medium and 10% Cultured in fetal bovine serum at 37°C, 5% CO 2 in the incubator.

[0068] 2. QPCR

[0069] 2.1 Extraction of total cellular RNA: Extraction of total cellular RNA was performed using the RNA extraction kit from QINGEN Company, and performed according to the instruction manual.

[0070] 2.2 QPCR: the steps are the same as in Example 2.

[0071] 3. Results

[0072] Such as figure 2 As shown, compared with the human lung epithelial cell line BEAS-2B, the expression of miRNA-2116 in lung adenocarcinoma cell lines A549, NCI-H1650, NCI-H1299, and SPC-A-1 was significantly increased (P<0.05).

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Abstract

The invention discloses a miRNA marker, in particular to a lung adenocarcinoma miRNA marker which is miRNA-2116. The miRNA-2116 can be used for judging whether lung adenocarcinoma metastasis happens or predicting the metastasis risk of the lung adenocarcinoma. Test results show that the miRNA-2116 can effectively distinguish a lung adenocarcinoma metastasis sample from a non-metastasis sample. On the basis, the miRNA-2116 can also be used for preparing medicines for restraining lung adenocarcinoma metastasis or preventing lung adenocarcinoma metastasis. The lung adenocarcinoma miRNA marker provides a new diagnostic method for diagnosing lung adenocarcinoma metastasis at the molecular level clinically, and provides a new medicine target for gene therapy of lung adenocarcinoma metastasis.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a lung adenocarcinoma miRNA marker and its application, in particular to a miRNA-2116 marker related to lung adenocarcinoma metastasis and its application. Background technique [0002] miRNA is a 21-22nt non-coding RNA molecule that naturally exists in the body. It is a class of RNA that regulates the expression of target genes through post-transcriptional gene silencing. It is estimated that about 1 / 3 of genes in organisms are regulated by miRNAs. The complex of miRNA and RISC can bind to the complementary sequence in the 5'-UTR or 3'-UTR of the target gene mRNA through base pairing, inhibit protein translation, or trigger mRNA degradation, thereby negatively regulating the expression of the target gene. [0003] Lung cancer is the cancer with the highest mortality rate, with more than 1 million deaths and 1.2 million new cases each year. Lung adenocarcinoma (lung adenocarcinoma) is t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B40/06A61K45/00A61P35/00
CPCA61K48/00C12Q1/6869C12Q1/6886C12Q2600/178C40B40/06
Inventor 杨承刚宋宏涛边洋
Owner 昆明博奥三合医学检验实验室有限公司
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