Application of molecular target in prognosis prediction and treatment of esophageal squamous cell carcinoma

A technique for esophageal squamous cell carcinoma and prognosis evaluation, applied in the field of biomedicine, can solve the problems of poor prognosis of esophageal cancer, inability to evaluate the prognosis with molecular markers, lack of accurate detection and treatment of esophageal squamous cell carcinoma, etc., and achieve the effect of accurate detection

Inactive Publication Date: 2019-05-10
SUN YAT SEN UNIV CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a combination of surgery, radiotherapy, chemotherapy, and biological therapy has been used in recent years, the prognosis of esophageal cancer is still poor, and the overall 5-year survival rate is less than 30%.
Although this method can help diagnose esophageal squamous cell carcinoma, the detection of CCKBR gene does not provide an effective therapeutic target for esophageal squamous cell

Method used

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  • Application of molecular target in prognosis prediction and treatment of esophageal squamous cell carcinoma
  • Application of molecular target in prognosis prediction and treatment of esophageal squamous cell carcinoma
  • Application of molecular target in prognosis prediction and treatment of esophageal squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Screening lncRNAs that play a key role in the occurrence and development of esophageal squamous cell carcinoma

[0047] 1.1 TCGA database screening

[0048] Using bioinformatics technology to screen lncRNAs highly expressed in esophageal squamous cell carcinoma from the TCGA database.

[0049] 1.2 siRNA library construction and screening

[0050] Using the top 50 highly expressed LncRNAs in cancer tissues analyzed from the database, an siRNA library was established and transfected into esophageal cancer cell lines EC9706 and KYSE30. By detecting MTS and lactic acid production, LncRNAs that can significantly affect cell proliferation and glucolysis were observed.

[0051] 1.3 Large sample tissue verification

[0052] The TCGA database was used to analyze the expression level of lncRNA APGG in large samples of cancers such as esophageal cancer, gastric cancer, and colorectal cancer. The analysis results are as follows: figure 1 shown by figure 1 It can be se...

Embodiment 2

[0053] Example 2 uses the kit to verify the expression level of lncRNA APGG through RT-PCR technology

[0054] 2.1 Acquisition of cDNA

[0055] RNA extraction: Take relevant samples of esophageal cancer (ESCC), gastric cancer (GC), and colon cancer (CRC) tissues. These samples have obtained the consent of relevant personnel, and the total RNA of animal tissues is extracted using RNA extraction kit NEP019-1, and then Use Bio-Red ultraviolet spectrophotometer to measure the optical density value of total RNA at 280nm and 260nm, measure OD 260 / OD 280 The value is 1.87, indicating that the RNA purity is reliable and can be used for the next test,

[0056] Acquisition of cDNA: Use the Revert Aid First Strand cDNA synthesis Kit to reverse transcribe RNA into cDNA;

[0057] 2.2 Primer design

[0058] The sequence information of the APGG gene (that is, LOC148709) was screened out from the TCGA database, and the amplification primers for RT-PCR were designed using PrimerPrimier5.0...

Embodiment 3

[0065] Example 3 Using RNA scope in situ hybridization technology to verify the expression level of lncRNA APGG

[0066] RNAscope is a new type of ultra-sensitive RNA in situ hybridization detection technology. It has a unique specific probe design and signal amplification system, which can extremely reduce background noise. Its main operation steps include: (1) Pretreatment of tissue sections to increase Its permeability; (2) target RNA hybridizes with specific probe; (3) signal amplification system hybridizes with probe; (4) observes the result under bright field or fluorescent microscope.

[0067] For the specific results of APGG, see image 3 ,Depend on image 3 It can be seen that lncRNA APGG is highly expressed in cancer tissues such as esophageal cancer, gastric cancer, and colorectal cancer.

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Abstract

The invention belongs to the technical field of biomedicine, and particularly relates to the application of a molecular target in the prognosis prediction and treatment of esophageal squamous cell carcinoma. The molecular target is a long-chain non-coded RNA AGPG. The expression level of an AGPG gene in esophageal squamous cell carcinoma tissues is detected through RT-PCR, RNA scope in-situ hybridization, in-situ hybridization, a chip or a high-throughput sequencing platform. The expression level of a target protein PFKFB3 corresponding to the AGPG gene in the esophageal squamous cell carcinoma tissues is detected through immunoblot and an immunohistochemical technology. A biologically active drug is successfully developed, the biologically active drug comprises an inhibitor of the long-chain non-coded RNA AGPG and an inhibitor PFK15 of a PFKFB3 protein, and the biologically active drug can be effectively used for the prognosis prediction and treatment of the esophageal squamous cell carcinoma. A novel molecular target is provided for the prognosis prediction and treatment of the esophageal squamous cell carcinoma by a researching outcome, and a novel treatment strategy is providedfor the clinical treatment of the esophageal squamous cell carcinoma.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of a molecular target in the prognosis evaluation and treatment of esophageal squamous cell carcinoma. Background technique [0002] Esophageal cancer is one of the most common malignant tumors in humans, ranking ninth in the incidence of tumors in the world, and ranking eighth in mortality. Esophageal cancer is a high-incidence tumor in my country, ranking fourth in cancer mortality. Although a combination of surgery, radiotherapy, chemotherapy, and biological therapy has been used in recent years, the prognosis of esophageal cancer is still poor, and the overall 5-year survival rate is less than 30%. 90% of the pathological types of esophageal cancer are squamous cell carcinomas, especially in Asia, where esophageal squamous cell carcinoma is the most important pathological type. Currently, esophageal squamous cell carcinoma lacks effective pro...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/574A61K45/00A61P35/00
Inventor 徐瑞华鞠怀强刘泽先刘佳
Owner SUN YAT SEN UNIV CANCER CENT
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