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Application of circ_2157 in preparation of therapeutic preparation for nasopharyngeal carcinoma and therapeutic preparation

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

Active Publication Date: 2019-05-17
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_2157 in preparation of therapeutic preparation for nasopharyngeal carcinoma and therapeutic preparation
  • Application of circ_2157 in preparation of therapeutic preparation for nasopharyngeal carcinoma and therapeutic preparation
  • Application of circ_2157 in preparation of therapeutic preparation for nasopharyngeal carcinoma and therapeutic preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1: Expression of circ_2157 in nasopharyngeal carcinoma tissues and cells

[0086] 1. According to the standard sample collection plan, we collected 41 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). 29 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 permission...

Embodiment 2

[0124] Example 2: Detection of overexpression effect of circ_2157 in nasopharyngeal carcinoma cell lines

[0125] First, we selected the restriction site, put the full-length sequence of circ_2157 into the NEB cutter 2.0 online website for analysis, and showed that the ClaI and SacII restriction sites were sites that did not exist in the full-length sequence of circ_2157, and at the same time in the pcDNA3.1 plasmid vector ( Purchased from Invitrogen Corporation) DNA restriction endonuclease that exists alone. Construct an overexpression vector accordingly; figure 2 The overexpression vector map drawn for.

[0126] In order to test the circling efficiency of circ_2157, we first expressed the constructed pcDNA3.1 / circ_2157 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

[0127] Example 3: Detection of the effect of silencing circ_2157 expression in nasopharyngeal carcinoma cell lines

[0128]The SI sequence of circ_2157 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_2157 in tumorigenesis and development, we designed siRNA according to the splicing site of circ_2157, and transiently transfected siRNA and siNC (blank control) into HONE1, HNE2 and CNE2 cell lines with Hiperfect reagent to silence the expression of circ_2157. 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_2157 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 application of circ_2157 in preparation of a therapeutic preparation for nasopharyngeal carcinoma and the therapeutic preparation. Since siRNA has a good silencing effect, an MTT assay is performed in nasopharyngeal carcinoma cell lines HONE1, HNE2 and CNE2 in which the circ_2157 is silenced afterthe circ_2157 is disturbed, and the proliferation rate of cells in the siRNA group is significantly lower than that in the NC group, that is, the silenced circ_2157 inhibits the proliferation of nasopharyngeal carcinoma cells. That is, inhibition of the circ_2157 can treat the nasopharyngeal carcinoma, and the circ_2157 has far-reaching clinical significance and important 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_2157 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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IPC IPC(8): A61K31/713A61P35/00
Inventor 曾朝阳郭灿熊炜王忆安李桂源李小玲莫勇真龚朝建张姗姗周鸣周艳宏李征
Owner CENT SOUTH UNIV
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