Application of circ_ADARB1 in preparing nasopharyngeal carcinoma therapeutic preparation and therapeutic preparation

A technology for nasopharyngeal carcinoma and preparations, which is applied in the field of tumor molecular biology and can solve the problems of unclear molecular mechanism and insignificant curative effect.

Active Publication Date: 2019-06-21
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the specific molecular mechanism of the occurrence and development of nasopharyngeal carcinoma is still unclear, and due to tum

Method used

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  • Application of circ_ADARB1 in preparing nasopharyngeal carcinoma therapeutic preparation and therapeutic preparation
  • Application of circ_ADARB1 in preparing nasopharyngeal carcinoma therapeutic preparation and therapeutic preparation
  • Application of circ_ADARB1 in preparing nasopharyngeal carcinoma therapeutic preparation and therapeutic preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1: Expression of circ_ADARB1 in nasopharyngeal carcinoma tissues and cells

[0092] 1. According to the standard sample collection plan, we collected 34 tissue samples from patients with nasopharyngeal carcinoma from Hunan Cancer Hospital. All the cases were newly diagnosed patients in the Department of Head and Neck Surgery of Hunan Cancer Hospital (time interval: January 2016 to November 2016). 22 cases of nasopharyngeal carcinoma and 12 cases of nasopharyngeal inflammation were diagnosed by the pathology department (neoplastic diseases, no active infectious diseases, severe immune diseases and other major diseases have been excluded).

[0093] Complete personal information and clinical data were recorded during the collection process, including name, gender, age, outpatient number, hospitalization number, pathological type, case stage, and EBV infection, etc. See the screenshot of the Excel spreadsheet for details. All samples are collected with the permissi...

Embodiment 2

[0129] Example 2: Sanger sequencing proves that circular RNA is formed

[0130] In order to prove that circ_ADARB1 forms a circular RNA rather than a linear one, the figure 1 The qRT-PCR products were recovered and sent to the company for sanger sequencing (Qingke Company). Compare the sequences returned by the company with DNASTAR software, and use the chromas software to view the peak diagrams to judge the quality of the sequencing. figure 2 The results in the middle show that a. circ_ADARB1 is formed by end-to-end splicing of exons 2 and 3 of ARHGAP12, E indicates exon (exon), and the underlined sequence indicates the linker sequence at the end; b. Schematic diagram of circRNA formation; c. Peak of sequencing results In the figure, the black arrow indicates the end-to-end connection from here.

Embodiment 3

[0131] Example 3: Detection of the overexpression effect of circ_ADARB1 in nasopharyngeal carcinoma cell lines

[0132]First, we select the restriction site, and put the full-length sequence of circ_ADARB1 into the NEB cutter 2.0 online website for analysis. It shows that the ClaI and SacII restriction sites are sites that do not exist in the full-length sequence of circ_ADARB1, and at the same time in the pcDNA3.1 plasmid vector A single DNA restriction enzyme. Construct the overexpression body accordingly; image 3 The overexpression vector map drawn for.

[0133] In order to test the circling efficiency of circ_ADARB1, we first expressed the constructed pcDNA3.1 / circ_ADARB1 eukaryotic overexpression vector in nasopharyngeal carcinoma cells. The third and fourth generation nasopharyngeal carcinoma cells CNE2 and HNE2 in good growth condition were seeded into 12-well plates, and when the cell confluence reached 60%-80%, the endotoxin-free plasmid pcDNA3. 1 Empty vector and...

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Abstract

The invention belongs to the technical field of tumor molecular biology, in particular to an application of circ_ADARB1 in preparing a nasopharyngeal carcinoma therapeutic preparation and a therapeutic preparation. For the first time, we found that circ_ADARB1 may become a potential target for nasopharyngeal carcinoma treatment. As the siRNA has a good silencing effect, the siRNA designed for a splicing site at the head-tail junction of the circular RNA is adopted to interfere with circ_ADARB1, scratch healing experiments and matrix gel invasion experiments are carried out in nasopharyngeal carcinoma cell lines HNE2 and CNE2, compared with a control group, the invasion and migration ability of cells in circ_ADARB1 group knocked down by the siRNA are obviously weakened, namely, the silencing circ_ADARB1 inhibits the invasion and metastasis of nasopharyngeal carcinoma cells. The application shows that inhibition of circ_ADARB1 can treat nasopharyngeal carcinoma and has far-reaching clinical significance and important popularization and application prospects.

Description

technical field [0001] The invention belongs to the technical field of tumor molecular biology, and in particular relates to a reagent for inhibiting circular RNA circ_ADARB1 and its application in preparing nasopharyngeal carcinoma treatment preparations. Background technique [0002] Circular RNA (Circular RNA) is a major research hotspot. They are a type of reverse splicing of precursor mRNA (precursormessenger RNA, pre-mRNA) without a 5' end cap and a 3' end poly(A ) tail and covalently bonded to form a non-coding RNA molecule in a circular structure. It has the characteristics of high stability, conservation, specificity, and high content. CircRNAs are mainly derived from exon regions of protein-coding genes, and can also be formed from intron regions, UTR regions, intergenic regions, non-coding RNA sites, and antisense sites of known transcripts. [0003] The formation process of circRNA can be divided into two categories, namely, exon circularization and intron circ...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/713A61P35/00
Inventor 熊炜郭灿曾朝阳唐乐李桂源李小玲魏芳武迎芬伍旭周鸣莫勇真范春梅
Owner CENT SOUTH UNIV
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