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Application of circ_3480 to preparation of treatment preparation for nasopharyngeal carcinoma and treatment preparation

A nasopharyngeal carcinoma and preparation technology, applied in the field of tumor molecular biology, can solve problems such as unclear molecular mechanism and insignificant curative effect

Active Publication Date: 2019-06-07
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 tumor heterogeneity and individual differences, the curative effect of traditional radiotherapy combined with chemotherapy is not significant

Method used

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  • Application of circ_3480 to preparation of treatment preparation for nasopharyngeal carcinoma and treatment preparation
  • Application of circ_3480 to preparation of treatment preparation for nasopharyngeal carcinoma and treatment preparation
  • Application of circ_3480 to preparation of treatment preparation for nasopharyngeal carcinoma and treatment preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1: Expression of circ_3480 in nasopharyngeal carcinoma tissues and cells

[0086] 1. According to the standard sample collection plan, we collected 40 tissue samples from patients with nasopharyngeal carcinoma from Hunan Provincial 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). 28 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).

[0087] 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...

Embodiment 2

[0123] Example 2: Detection of overexpression effect of circ_3480 in nasopharyngeal carcinoma cell lines

[0124] First, we selected the restriction site, and put the full-length sequence of circ_3480 into the NEB cutter 2.0 online website for analysis. It showed that the ClaI and SacII restriction sites were sites that did not exist in the full-length sequence of circ_3480. At the same time, in the pcDNA3.1 plasmid vector ( Purchased from Invitrogene Corporation) DNA restriction endonuclease that exists alone. Construct an overexpression vector accordingly; figure 2 The overexpression vector map drawn for.

[0125] In order to test the circling efficiency of circ_3480, we first expressed the constructed pcDNA3.1 / circ_3480 eukaryotic overexpression vector in nasopharyngeal carcinoma cells. The third and fourth generation nasopharyngeal carcinoma cells HONE1, HNE2 and CNE2 in good growth condition were seeded into 12-well plates, and when the cell confluence reached 60%-80%,...

Embodiment 3

[0126] Example 3: Detection of the effect of silencing circ_3480 expression in nasopharyngeal carcinoma cell lines

[0127]The SI sequence of circ_3480 was designed according to the splicing site. siRNA is a kind of double-stranded RNA molecule with a length of 21-25 nucleotides, which can complementarily bind with homologous RNA, specifically degrade the target RNA, and thereby inhibit its expression. At present, siRNA has developed into an important tool for the study of gene function. In order to explore the role of circ_3480 in tumorigenesis and development, we designed siRNA according to the splicing site of circ_3480, and transiently transfected siRNA and siNC (blank control) into HONE1, HNE2 and CNE2 cell lines with Hiperfect reagent to silence the expression of circ_3480. Continue to culture for 48 hours after transfection to collect cells, use real-time fluorescent quantitative PCR technology to detect the expression level of circ_3480 to detect the transfection effic...

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Abstract

The invention belongs to the technical field of tumor molecular biology, and particularly relates to an application of circ_3480 to preparation of a treatment preparation for nasopharyngeal carcinomaand the treatment preparation. The siRNA has favorable silencing effects, so that after the situation that the circ_3480 is in interference is guaranteed, MTT experiment is performed in nasopharyngealcarcinoma cell lines HONE1, HNE2 and CNE2 silencing the circ_3480; and relative to an NC group, a siRNA group has the advantages that the cell proliferation speed is obviously reduced, namely that when the circ_3480 is silenced, the cell proliferation of the nasopharyngeal carcinoma can be restrained, and when the circ_3480 is restrained, the nasopharyngeal carcinoma can be treated. The circ_3480has far-reaching clinical significance and important promotion 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_3480 and its application in the preparation of nasopharyngeal carcinoma treatment preparations. Background technique [0002] At present, circular RNA (circular RNA) is a research hotspot. CircRNA is mainly derived from the exon region of protein-coding genes, and can also be composed of intron regions, UTR regions, intergenic regions, non-coding RNA sites and known transcripts. antisense site formation. CircRNA is a kind of non-coding RNA formed by reverse splicing of precursor mRNA (precursor messenger RNA, pre-mRNA) without a 5' end cap and a 3' end poly(A) tail, and forms a circular structure with a covalent bond. molecular. [0003] The formation process of circRNA can be divided into two categories, namely exon circularization (exon circularization) and intron circularization (intron circularization). ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/713A61P11/02A61P11/04A61P35/00
Inventor 熊炜曾朝阳郭灿王忆安李桂源李小玲伍旭范春梅唐艳艳龚朝建张姗姗周鸣
Owner CENT SOUTH UNIV
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