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Application of mir-26a in non-small cell lung cancer

A mir-26a and gene technology, applied in the application field of miR-26a in non-small cell lung cancer, can solve problems such as abnormal expression and complex function of miR-26a cells

Inactive Publication Date: 2020-09-08
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The current research results show that the cellular function of miR-26a is very complex, and it is abnormally expressed in a variety of disease cells

Method used

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  • Application of mir-26a in non-small cell lung cancer
  • Application of mir-26a in non-small cell lung cancer
  • Application of mir-26a in non-small cell lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Determining the expression level changes of miR-26a in mouse-induced NSCLC tissues

[0037] The gene mutant mouse lung cancer models (L822T1, L703T2, L903T1) and normal lung cancer model (L1805) used in the present invention were provided by the research group of Professor Ji Hongbin, Institute of Biochemical Cells, Chinese Academy of Sciences. L1805 is mouse normal lung tissue, L703T2 is p53 - / - / Kras + / + malignancy of L903T1 for Lkb1 - / - / Kras + / + malignancy of L822T1 for Kras + / + of benign tumors. The lung tissue of the mouse and the lung tissue of the induced cancer were sampled separately, and the total RNA in the tissue was extracted using the Trizol reagent provided by TaKaRa Company, and the reverse transcription was performed using the One Step PrimeScript miRNA cDNASynthesis Kit (Code No. D350A) of TaKaRa Company , construct the cDNA library of the above tissue, use oligo dt as a primer for reverse transcription, and obtain the miRNA cDNA libra...

Embodiment 2

[0040] Example 2: Study on the expression level changes of miR-26a in human NSCLC cells and tissues

[0041] According to the sequencing results of mouse tumor samples, it can be guessed that miR-26a is a cytokine related to lung cancer. In order to determine whether this conclusion is also applicable to humans, we detected the expression of miR-26a in human lung cancer cells by qRT-PCR. expression levels in the system.

[0042] The cell lines used in the present invention include: HCC827 is EGFR + / + Non-small cell lung cancer cell line, A549 for Kras + / + Non-small cell lung cancer cell line, H1299 for p53 - / - Non-small cell lung cancer cell line, SPCA-1 is an Asian non-small cell lung cancer cell line, 95-D is a human metastatic large cell lung cancer cell line, BEAS-2B is a human lung epithelial cell line. The 24 NSCLC tissue samples and corresponding paracancerous tissues used in the present invention are all from Shanghai Chest Hospital.

[0043] In this example, the T...

Embodiment 3

[0047] Example 3: The effect of miR-26a on the proliferation of NSCLC cells

[0048] The detection of cell proliferation ability was completed using CCK-8 assay and clone formation assay. The specific operation steps are as follows:

[0049] Cell Counting Kit-8 (CCK-8 for short) is a kit for detecting cell proliferation and cytotoxicity developed by DOJINDO, Japan. The specific operation steps of CCK-8 detection are as follows: take the cells in the logarithmic growth phase, seed them in 96-well cell culture plates at a density of 2000 / well, and store them at 37°C, 5% CO 2 cultivated under conditions. At different time points (24, 48, 72, 96 h), 5 µl of CCK-8 solution was added to each well at 37°C, 5% CO 2 After incubation for 1 h, the absorbance at 450 nm was detected using a microplate reader. In order to ensure the reliability of the experiment, three auxiliary holes were set for each group of samples at the same time. In addition, in order to slow down the evaporation...

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Abstract

The invention relates to an application of miR-26a in non-small cell lung cancer (NSCLC). MiR-26a can effectively inhibit the proliferation of NSCLC cells, and promote the apoptosis of lung cancer cells. MiRNA target gene prediction software shows that WNK3 is a potential target spot of miR-26a, the 3'-UTR area of WNK3 has two binding sites in complementary pairing with the seed sequence of miR-26a. The test shows that miR-26a can inhibit the translation and expression of target genes in the protein level. A bifluorescence reporter gene system proves the direct targeted relationship of miR-26a and WNK3. The in-depth study shows that miR-26a opens the Caspase path of cell apoptosis by targeting WNK3 protein, and finally plays the role of inducing cell apoptosis, thus providing an important therapeutic target for the accurate treatment. Through the application of miR-26a in NSCLC, an action mechanism of miR-26a is verified, potential miRNA molecules with an anti-cancer function are provided clinically, a theoretical basis is provided for the study on the relevant drug targets, and novel accurate treatment research direction and application guidance are also provided.

Description

technical field [0001] The invention relates to an application of miR-26a in non-small cell lung cancer. Background technique [0002] Lung cancer is one of the most common malignant tumors with the highest mortality rate and the greatest threat to human health and life in the world. Lung cancer can be divided into small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). In my country, about 80% of lung cancers are diagnosed as NSCLC, which mainly includes adenocarcinoma, squamous cell carcinoma, and large cell undifferentiated carcinoma. Most lung cancer patients have no obvious symptoms in the early stage of the disease, which leads to late diagnosis, poor treatment effect, and low disease prognosis. Despite the great development of treatment methods, the five-year survival rate of NSCLC is still not higher than 15%. [0003] MicroRNA (microRNA, miRNA) is a class of non-coding small RNA that can regulate cellular functions. The length of miRNA is about 20-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K31/713A61P35/00
CPCA61K31/713
Inventor 金由辛金言袁天蔚李雪党启
Owner SHANGHAI UNIV
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