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Application of circular RNA circBRD7 in preparation of preparation for diagnosing and/or treating nasopharyngeal carcinoma

A nasopharyngeal carcinoma and preparation technology, which is applied in the field of preparations for the diagnosis and treatment of nasopharyngeal carcinoma, can solve the problems of concealed disease location, low survival rate of nasopharyngeal carcinoma patients, resistance to radiotherapy and chemotherapy, etc.

Pending Publication Date: 2022-08-09
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although clinical and experimental oncology has made great progress in the diagnosis of nasopharyngeal carcinoma in recent years, the prognosis of nasopharyngeal carcinoma patients has not been significantly improved, and local recurrence and distant metastasis are still the low survival rate of advanced nasopharyngeal carcinoma patients. The main reason
Patients with early nasopharyngeal carcinoma are very sensitive to radiotherapy and chemotherapy, and have very good clinical efficacy; however, due to the hidden location of the disease and atypical early symptoms, many nasopharyngeal carcinoma patients are diagnosed at a late stage when they are first diagnosed, and miss the best clinical treatment stage, thus prone to chemoradiotherapy resistance and distant metastasis
Therefore, early diagnosis and individualized treatment of nasopharyngeal carcinoma are very critical in the clinical diagnosis and treatment of nasopharyngeal carcinoma. However, there are no screening markers for early diagnosis and progress evaluation of nasopharyngeal carcinoma clinically. A diagnostic method for early diagnosis and progression evaluation of nasopharyngeal carcinoma

Method used

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  • Application of circular RNA circBRD7 in preparation of preparation for diagnosing and/or treating nasopharyngeal carcinoma
  • Application of circular RNA circBRD7 in preparation of preparation for diagnosing and/or treating nasopharyngeal carcinoma
  • Application of circular RNA circBRD7 in preparation of preparation for diagnosing and/or treating nasopharyngeal carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1, confirming the real existence of circBRD7 in nasopharyngeal carcinoma cells

[0058] 1. Materials and methods:

[0059] 1.1 Reagents and kits

[0060] Agarose (agrose), nucleic acid dyes, and DL2000 Marker and other commonly used biochemical reagents for agarose gel electrophoresis were purchased from US EVERBRIGHT Company; Actinomycin D (Dactinomycin) was purchased from Selleck Company; Cytoplasmic and Nuclear RNA Isolation Kit (PARIS) TM Kit Protein and RNA Isolation System) was purchased from Invitrogen Company. Use AG RNAex Pro Reagent (Aike Rui company) to pump RNA of improved quality, IIQ RT SuperMix for qPCR (+gDNA wiper) reverse transcription kit (Vazyme) reverse transcribed RNA into cDNA, and ChamQ Universal SYBR qPCR Master Mix qPCR kit was used for real-time fluorescence quantitative PCR detection. The nasopharyngeal carcinoma cell lines (NP69, CNE1, CNE2, HNE1, HNE2, HONE1, 5-8F, HK1, 6-10B, C666-1) used in the present invention are preserved...

Embodiment 2

[0174] Example 2. In situ hybridization to detect the expression of circBRD7 in nasopharyngeal carcinoma

[0175] 1. Materials and methods:

[0176] 1.1 Design and synthesis of hybridization probes

[0177] In order to detect the expression of circBRD7 in nasopharyngeal carcinoma patient tissues by in situ hybridization, we designed an oligonucleotide probe for detecting circBRD7 expression at the circRNA splice site and a positive control in situ hybridization oligonucleotide for detecting GAPDH. nucleotide probes.

[0178] Oligonucleotide probes for in situ hybridization detection of circBRD7 expression:

[0179] Preferred: circBRD7 probe:

[0180] 5’-TAGTTTGAAGTTATCCTGACTGTTCACAAG-3’

[0181] A positive control probe for detecting the internal reference gene GAPDH (preferably any of the following):

[0182] Probe sequence 1: 5'-CAGTAGAGGCAGGGATGATGTTCTGGAGAG-3'

[0183] Probe sequence 2: 5'-CCACTTTACCAGAGTAAAAGCAGCCCTGG-3'

[0184] The above-mentioned oligonucleotide...

Embodiment 3

[0233] Example 3. In vitro overexpression of circBRD7 inhibits the proliferation of nasopharyngeal carcinoma and induces apoptosis

[0234] 1. Materials and methods

[0235] 1.1 Reagents and kits

[0236] Restriction endonucleases ClaI and SacII were purchased from TakaRa company; T4 DNA ligase (PROMEGA); plasmid extraction kit (Tiangen); purification kit (OMEGA); Kit (Biomake Biological Company); Crystal Violet (Beijing Biyuntian Biological Company); TksGflex TM DNA Polymerase (Takara Company); AnnexinV-FITC / PI Kit (Sizhengbai Biotechnology). Use AG RNAex Pro Reagent (Aike Rui company) to pump RNA of improved quality, II Q RT SuperMix for qPCR (+gDNA wiper) reverse transcription kit (Vazyme) reverse-transcribes RNA into cDNA, and ChamQ Universal SYBR qPCR Master Mix qPCR kit performs real-time quantitative PCR detection.

[0237] 1.2 Construction of circBRD7 overexpression vector

[0238] (1) First, we selected the restriction sites and put the full-length sequence of ...

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Abstract

The invention discloses an application of circular RNA circBRD7 in preparation of a preparation for diagnosing and / or treating nasopharyngeal carcinoma. The circular RNA circBRD7 can be used for preparing a diagnosis and treatment preparation for a nasopharyngeal carcinoma patient, and particularly preparing a kit for diagnosing the nasopharyngeal carcinoma patient by an in-situ hybridization detection method. Researches prove that expression down-regulation of circBRD7 in nasopharyngeal carcinoma tissues and nasopharyngeal carcinoma cell lines is in negative correlation with clinical progress of nasopharyngeal carcinoma patients. Besides, the circBRD7 plays a role in tumor suppressor genes in nasopharyngeal carcinoma, and the defect of circBRD7 in nasopharyngeal carcinoma cells and tissues can be made up by exogenous overexpression of circBRD7, so that the purpose of reversing malignant phenotypes of nasopharyngeal carcinoma is achieved. Therefore, expression detection and exogenous overexpression based on circBRD7 can be used for early molecular diagnosis, progress evaluation and clinical targeted therapy of nasopharyngeal carcinoma patients, and has important clinical value and popularization and application prospects.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and in particular relates to an in situ hybridization probe of circular RNA circBRD7, an expression vector and a preparation of circBRD7 for the diagnosis and treatment of nasopharyngeal carcinoma. Background technique [0002] Nasopharyngeal carcinoma is a malignant head and neck tumor originating from nasopharyngeal epithelial cells, with obvious regional aggregation, high degree of malignancy, and strong invasive and metastatic ability. Although clinical and experimental oncology has made great progress in the diagnosis of nasopharyngeal carcinoma in recent years, the prognosis of nasopharyngeal carcinoma patients has not improved significantly, and local recurrence and distant metastasis are still the low survival rate of patients with advanced nasopharyngeal carcinoma. main reason. Early-stage nasopharyngeal cancer patients are very sensitive to radiotherapy and chemotherapy, and have...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6841C12N15/11A61K31/7088A61P35/00
CPCC12Q1/6886C12Q1/6841A61K31/7088A61P35/00C12Q2600/178C12Q2600/158C12Q2600/166C12Q2543/10C12Q2563/173
Inventor 周鸣魏建霞李梦娜陈峙朋薛长宁段玉梅郑乐媚李桂源熊炜曾朝阳李小玲
Owner CENT SOUTH UNIV