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Function of HOXD-AS1 in diagnosis and treatment of esophageal squamous carcinoma

A technology for esophageal cancer and esophageal squamous cells, applied in the field of tumor molecular biology, can solve the problem of less functional research

Active Publication Date: 2014-07-16
江苏万成生物医学研究院有限公司
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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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  • Function of HOXD-AS1 in diagnosis and treatment of esophageal squamous carcinoma
  • Function of HOXD-AS1 in diagnosis and treatment of esophageal squamous carcinoma
  • Function of HOXD-AS1 in diagnosis and treatment of esophageal squamous carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 lncRNA chip expression analysis of human esophageal squamous cell carcinoma and paired normal tissues

[0055] 1. Materials and methods

[0056] 1. Materials

[0057] 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.

[0058] 2. Method

[0059] 2.1 Extraction of total RNA from tumor tissue and paired normal tissue

[0060] 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

[0083] Example 2 qRT-PCR preliminary verification of differential expression of HUESCC-lncRNA5 in cancer tissues of esophageal squamous cell carcinoma and paired normal tissues

[0084] 1. Experimental materials

[0085] Another 30 pairs (different from the samples tested by the microarray) of human esophageal squamous cell carcinoma tissues and paired normal tissues were selected, and the expression difference of HUESCC-lncRNA5 was initially verified by qRT-PCR.

[0086] 2. Experimental methods and results

[0087] 1 Identification of primer specificity

[0088] 1.1 Screening of specific primers

[0089] (1) Extract the transcript sequence related to HUESCC-lncRNA5 from the Ensemble database, and use the primer design tool (Primer-BLAST) of NCBI to design primers according to the sequence of the transcript;

[0090] (2) The designed primers were evaluated by Oligo7, and 3 pairs of primers were designed for each;

[0091] Pair1 upstream primer: SEQ ID NO.2

[0092] Pair1 ...

Embodiment 3

[0135] Example 3 qRT-PCR further verified the differential expression of HUESCC-lncRNA5 in cancer tissues and paired normal tissues of esophageal squamous cell carcinoma

[0136] 1. qRT-PCR kit composition

[0137] 1.1 Composition of dye-based HUESCC-lncRNA5qRT-PCR kit:

[0138] (1) Upstream primer: SEQ ID NO.4

[0139] (2) Downstream primer: SEQ ID NO.5;

[0140] Other reagents refer to SYBR Premix Ex Taq TM II (Tli RNaseH Plus) fluorescence quantitative kit (Code No.RR820A).

[0141] 2. Detection of HUESCC-lncRNA5qRT-PCR

[0142] 2.1 Preparation of total RNA

[0143] 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 ND-1000 Nucleic Acid...

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Abstract

The invention discloses a function of HOXD-AS1 in diagnosis and treatment of esophageal squamous carcinoma and relates to discovery, detection and application of long-chain noncoding RNA. According to the sequences of the long-chain noncoding RNA, a specific real-time quantitative PCR primer is designed and synthesized; a real-time quantitative PCR preparation is utilized to detect the expression level of the long-chain noncoding RNA in a clinical case specimen of the esophageal squamous carcinoma, and results show that the expression of the long-chain noncoding RNA in the esophageal squamous carcinoma is remarkably up-regulated and the expression level of the long-chain noncoding RNA is highly associated with both clinical stages and pathological N stages of an esophageal squamous carcinoma patient. The specific detection sequences disclosed by the invention are expected to be used for preparing the preparation for auxiliary diagnosis, curative effect prediction or prognosis judgment of the esophageal squamous carcinoma. In addition, two siRNA sequences for specifically knocking down the expression of the long-chain noncoding RNA are designed and synthesized; an experiment proves that after the expression level of the long-chain noncoding RNA is specifically knocked down in a human esophageal squamous carcinoma cell strain, the quantity of migrated tumor cells is remarkably reduced; therefore the siRNA sequences disclosed by the invention is also expected to be used as a molecular targeting treatment tool for inhibiting the cell migration rate of the esophageal squamous carcinoma.

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): C12Q1/68C12N15/113A61P35/00
CPCC12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 李苏卿曹秀峰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 江苏万成生物医学研究院有限公司
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