Detection and application of esophageal carcinoma-related long non-coding RNA

A long-chain non-coding, esophageal cancer technology, applied in the field of tumor molecular biology, can solve the problem of less functional research

Inactive Publication Date: 2014-07-30
NANJING FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, following the literature, it is found that except for the inventor's research group, there are still few studies on the lncRNA expression profile of esophageal squamous cell carcinoma and the functionality of related indicators.

Method used

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  • Detection and application of esophageal carcinoma-related long non-coding RNA
  • Detection and application of esophageal carcinoma-related long non-coding RNA
  • Detection and application of esophageal carcinoma-related long non-coding RNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 lncRNA chip expression analysis of human esophageal squamous cell carcinoma and paired normal tissues

[0050] 1. Materials and methods

[0051] 1. Materials

[0052] Tissue samples were obtained from inpatient surgical resection samples of 6 pairs of patients with esophageal squamous cell carcinoma, each pair containing esophageal tumor tissue and paired normal esophageal tissue.

[0053] 2. Method

[0054] 2.1 Extraction of total RNA from tumor tissue and paired normal tissue

[0055] According to Qiagen's RNA extraction kit (RNeasy Micro Kit, Cat. No. 74004), the total RNA was extracted from the tumor tissues of esophageal phosphocarcinoma patients and the paired normal tissues. The kit contains RNase-Free DNase I (lyophilized), which can effectively remove the impurities of genomic DNA. The purity and concentration of the extracted RNA were quantified by NanoDrop ND-1000 Nucleic Acid Quantifier (NanoDrop Technologies, Wilmington, Delaware), and the qua...

Embodiment 2

[0078] Example 2 Preliminary verification of differential expression of HDESCC-lncRNA3 in cancer tissue and normal tissue of esophageal squamous cell carcinoma by qRT-PCR

[0079] 1. Experimental materials

[0080] Another 30 pairs (different from the samples tested by the microarray) of human esophageal squamous cell carcinoma tissues and paired normal tissues were selected to conduct preliminary verification of the expression difference of HDESCC-lncRNA3 by qRT-PCR.

[0081] 2. Experimental methods and results

[0082] 1 Identification of primer specificity

[0083] 1.1 Screening of specific primers

[0084] (1) Extract the transcript sequence related to HDESCC-lncRNA3 from the Ensemble database, and use the primer design tool (Primer-BLAST) of NCBI to design primers according to the sequence of the transcript;

[0085] (2) The designed primers were evaluated by Oligo7, and 3 pairs of primers were designed for each;

[0086] Pair1 upstream primer: SEQ ID NO.2

[0087] P...

Embodiment 3

[0129] Example 3 qRT-PCR further verified the differential expression of HDESCC-lncRNA3 in cancer tissues and normal tissues of esophageal squamous cell carcinoma

[0130] 1. qRT-PCR kit composition

[0131] 1.1 Composition of dye-based HDESCC-lncRNA3qRT-PCR kit:

[0132] (1) Upstream primer: 5'-ATCG-3', SEQ ID NO.2

[0133] (2) Downstream primer: 5'-ATCG-3', SEQ ID NO.3;

[0134] Other reagents refer to SYBR Premix Ex Taq TM II (Tli RNaseH Plus) fluorescence quantitative kit (Code No.RR820A).

[0135] 2. Detection of HDESCC-lncRNA3qRT-PCR

[0136] 2.1 Preparation of total RNA

[0137] Another 80 pairs of cancer tissues and paired normal tissues of patients with esophageal squamous cell carcinoma were selected, according to the method of Life Technologies company. Reagents (Product No. 15596026) Reagents and steps required to extract total RNA, please refer to the instructions for details. The purity and concentration of the extracted RNA were quantified by NanoDrop N...

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Abstract

The invention relates to esophageal carcinoma-related long non-coding RNA and application thereof. Based on the sequence of the long non-coding RNA, specific qRT-PCR (Quantitative Real-time Polymerase Chain Reaction) primers and probes are designed and synthesized and reagents used for esophageal carcinoma auxiliary diagnosis or curative effect prediction are prepared. The qRT-PCR reagents are utilized for detecting the expression level of the long non-coding RNA in specimens of esophageal carcinoma clinical cases and a substantial decrease of the long non-coding RNS expression is found in esophageal carcinoma. The esophageal carcinoma-related long non-coding RNA can be applied to preparing reagents used for esophageal carcinoma auxiliary diagnosis, curative effect prediction or prognosis judgment.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and specifically relates to a long-chain non-coding RNA and its application. Specifically, the present invention relates to the application of a long-chain non-coding RNA in the preparation of preparations for auxiliary diagnosis or prognosis of esophageal cancer. Background technique [0002] Esophageal cancer (Esophageal Carcinoma, EC) is a malignant disease that seriously endangers the health of all human beings. Its morbidity and mortality rank the eighth and sixth respectively in the world. There are two main pathological types of EC: Esophageal Squamous Cell Carcinoma (ESCC) and Esophageal Adenocarcinoma (EAC). In my country, squamous cell carcinoma (90%) is the main pathological type. According to the latest data released by the World Health Organization, there are more than 250,000 new ESCC patients in my country every year, and about 200,000 patients die from esophageal cancer. , f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 曹秀峰李苏卿史卫红仝宇梭庹磊汪春梅谢海伟刘子豪杨同昕吕进纪律朱斌王和明李义生
Owner NANJING FIRST HOSPITAL
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