Application of miRNA-196b as non-small cell lung cancer molecular marker and treatment target

A 1. miRNA-196b, non-small cell lung cancer technology, applied in the fields of molecular biology and tumor prevention and treatment, can solve the problem that the mechanism of tumor suppression remains to be elucidated, and achieve the effect of improving tumor reproduction ability and promoting proliferation

Pending Publication Date: 2020-04-17
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] QKI-5 is a dominant subtype in non-small cell lung cancer (NSCLC), and the downregulation of QKI-5 is closely related to

Method used

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  • Application of miRNA-196b as non-small cell lung cancer molecular marker and treatment target
  • Application of miRNA-196b as non-small cell lung cancer molecular marker and treatment target
  • Application of miRNA-196b as non-small cell lung cancer molecular marker and treatment target

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Effect test

Embodiment 1

[0066] Example 1 The expression of QKI-5 is down-regulated in non-small cell lung cancer and is associated with the growth of lung cancer cells

[0067](1) In the TCGA RNA-seq database, the expression of QKI was analyzed in the tumor tissues of 334 patients with lung adenocarcinoma and their paired 57 normal paracancerous tissues, and the tumor tissues of 349 patients with lung squamous cell carcinoma and their matched pairs were also analyzed QKI expression in 51 cases of normal paracancerous tissues; QKI expression was significantly down-regulated in lung adenocarcinoma (p<2.2e-16) and lung squamous cell carcinoma (p<2.2e-16).

[0068] (2) Analyze the expression of QKI-5 in 30 pairs of non-small cell lung cancer (NSCLC) tissues and their corresponding adjacent tissues (NATs) from Zhoushan Hospital Affiliated to Wenzhou Medical University; Total RNA was extracted from 30 cases in each tissue, cDNA was synthesized, and then real-time fluorescent quantitative PCR was performed ...

Embodiment 2

[0073] Example 2 Down-regulation of QKI-5 promotes the expression of miR-196b

[0074] Nanostring nCounter miRNA expression analysis was performed with QKI-5 downregulated H1299 cells (H1299 / siQKI) and control H1299 (H1299 / siCont) cells.

[0075] The analysis method is as follows:

[0076] MicroRNA analysis was performed on total RNA extracted from QKI5 knockout H1299 cells and control cells using the NanoString nCounterhuman v3 miRNA Expression Assay following the manufacturer's instructions (NanoString Technologies). This assay allows the simultaneous detection and measurement of the expression levels of up to 800 different microRNAs per sample. Briefly, 3 μl of RNA is annealed to a multiplex DNA tag (miR-tag) and bridged to the specific target. Mature microRNAs are bound to specific miR-tags using ligase, followed by an enzymatic cleanup step to remove all excess tags. The labeled microRNA product was then diluted (1:5 ratio) and 5 μl was combined with 20 μl of reporter ...

Embodiment 3

[0081] Example 3 miR-196b promotes the growth of lung cancer cells both in vivo and in vitro

[0082] In the TCGA miRNA-seq database, the expression of miR-196b in 468 cases of lung adenocarcinoma, 198 cases of lung squamous cell carcinoma and 86 cases of NATs was analyzed. image 3 A) and lung squamous cell carcinoma ( image 3 B) Significantly upregulated.

[0083] Re-analysis of 70 pairs of NSCLC and NATs obtained from the shared resources of OSUCCC organizations confirmed that miR-196b expression was significantly up-regulated in NSCLC compared with the corresponding NATs ( image 3 C). Given the high expression of miR-196b in NSCLC tissues, we speculate that miR-196b plays an important role in the pathogenesis of NSCLC.

[0084] To explore the function of miR-196b in lung cancer cells, Pre-miR-196bPrecursor miRNA was transiently overexpressed in two NSCLC cell lines, H1299 and A549, because these two lines of cells have relatively low miR-196b compared to other lung ca...

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Abstract

The invention discloses an application of miRNA-196b as a non-small cell lung cancer molecular marker. It is found for the first time that cancer suppressor gene QKI-5 gene silencing can promote expression of miRNA-196b in lung cancer cells, and the expression of miR-196b can promote the proliferation, migration and cell cycle process of non-small cell lung cancer cells and improve the tumor reproductive capacity in animals by inhibiting the target genes GATA6 and TSPAN12 of the non-small cell lung cancer cells.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and tumor prevention and treatment, in particular to the application of miRNA-196b as a molecular marker and therapeutic target of non-small cell lung cancer. Background technique [0002] In my country, the morbidity and mortality of lung cancer rank first among malignant tumors for a long time, among which non-small cell lung cancer (NSCLC) is the most common, accounting for more than 80% of the total number of cases. With the advent of new chemotherapy drugs, targeted drugs and continuous improvement of surgical treatment techniques, the treatment effect of some lung cancer patients has been improved, but the 5-year survival rate is still only about 15%. The main reason is that the biological characteristics of lung cancer are very complex, the degree of malignancy is high, and due to the occultity of early symptoms, most lung cancer patients are already in the advanced stage when th...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/178C12Q2600/158C12Q2600/118
Inventor 崔日孟威竺王玉梁广黄翔杰
Owner WENZHOU MEDICAL UNIV
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