Application of gene HES2 to auxiliary diagnosis, prognosis and treatment of esophageal squamous carcinoma

A technology for esophageal squamous cell carcinoma and auxiliary diagnosis, which is applied in the fields of molecular biology and tumor drugs, and can solve the problems of limited tools for guiding treatment and immature markers of esophageal cancer

Active Publication Date: 2018-12-21
SUN YAT SEN UNIV CANCER CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, markers for predicting early recurrence and poor prognosis in esophageal cancer are immature, and tools to help new cases understand risk of disease progression and guide treatment are limited

Method used

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  • Application of gene HES2 to auxiliary diagnosis, prognosis and treatment of esophageal squamous carcinoma
  • Application of gene HES2 to auxiliary diagnosis, prognosis and treatment of esophageal squamous carcinoma
  • Application of gene HES2 to auxiliary diagnosis, prognosis and treatment of esophageal squamous carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1: HES2 is up-regulated in esophageal squamous cell carcinoma

[0075] (1) TCGA database analysis

[0076] Methods: In order to explore the expression of HES2 in esophageal cancer, the biological analysis method was used to analyze the mRNA expression data of Normal and Tumor in related esophageal cancer in The Cancer Genome Atlas (TCGA) database, and the prognosis analysis was carried out.

[0077] result( figure 1 A): The HES2 gene RNA level in esophageal squamous cell carcinoma tissue was higher than that in normal esophageal tissue and esophageal adenocarcinoma tissue.

[0078] (2) qRT-PCR detection of the mRNA level of HES2 gene in esophageal squamous cell carcinoma tissues and paired paracancerous tissues

[0079] Through the above analysis, it is known that the mRNA level of HES2 is highly expressed in esophageal squamous cell carcinoma, and the present invention further performs qRT-PCR detection and verification on HES2 in esophageal squamous cell car...

Embodiment 2

[0114] Example 2: Using small hairpin RNA (shRNA) to stably silence HES2 can suppress the malignant phenotype of esophageal cancer.

[0115] (1) Cell culture and stable strain construction

[0116] Methods: Esophageal cancer cells Eca109 and Kyse520 were treated with negative control shGFP and shRNA that can silence HES2 (including shHES2#1 and shHES2#2). WB verification was carried out by harvesting the corresponding protein. The targeting sequence of shHES2#1 is: 5'-CTGGAGCACCTGTGGCGGAG-3' (SEQ ID NO: 5), and the targeting sequence of shHES2#2 is: 5'-GCCTTGCGACAGCTACCGCG-3' (SEQ ID NO: 6).

[0117] Results: Control cells Eca109 shGFP, Kyse520 shGFP and HES2-silenced cells Eca109 shHES2#1, Eca109 shHES2#2, Kyse520 shHES2#1, Kyse520 shHES2#2 were obtained respectively.

[0118] (2) Cell growth experiment

[0119] Method: The culture medium containing 10% fetal calf serum was used to prepare a single cell suspension, and 1000-10000 cells per well were inoculated into a 96-we...

Embodiment 3

[0130] Example 3: Stable silencing of HES2 using antisense oligonucleotides (AON) targeting HES2 can inhibit the malignant phenotype of esophageal cancer

[0131] (1) Cell culture and stable strain construction

[0132] Methods: Esophageal cancer cells Eca109 were treated with negative control Scramble and AONs that could silence HES2 (HES2-AON#1, HES2-AON#2, HES2-AON#3, HES2-AON#4, HES2-AON#5), respectively, The working concentration is 5 nM.

[0133] HES2-AON #1: 5'-GCTCACTAGTCCACTA-3' (SEQ ID NO: 7);

[0134] HES2-AON#2: 5'-GCTGGTTAGATCTTAC-3' (SEQ ID NO: 8);

[0135] HES2-AON#3: 5'-GTGAAATTGATGCTCG-3' (SEQ ID NO: 9);

[0136] HES2-AON#4: 5'-AAGCAAGAGGTGGCAA-3' (SEQ ID NO: 10);

[0137] HES2-AON #5: 5'-TCCAGAGTCAAGCAAG-3' (SEQ ID NO: 11).

[0138] 1) qRT-PCR detection of HES2 mRNA expression level in esophageal squamous cell carcinoma cell lines after antisense oligonucleotide chain silenced HES2 gene.

[0139] Methods Six groups of cells were collected from the negat...

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Abstract

The invention discloses application of gene HES2 to the auxiliary diagnosis, prognosis and treatment of esophageal squamous carcinoma. The HES2 gene is highly expressed in esophageal squamous carcinoma tissues. The experiment further discovers that mRNA of HES2 gene is highly expressed in an esophageal squamous carcinoma cell line, the progress of the HES2 high-expressed tumor is faster and is more likely to be transferred to lymph gland or far organs, which is significantly related to poor five-year disease-free survival rate and five-year general survival rate. In addition, an antisense oligonucleotide chain sequence targeting HES2 gene is also designed to silencing HES2 expression and can be useor the trd featment of esophageal cancer. HES2 can be used as a detection marker for assisting the diagnosis of the esophageal squamous carcinoma and predicting the reoccurrence trend of the esophageal squamous carcinoma as well as a treatment target spot.

Description

technical field [0001] The invention relates to the fields of molecular biology and tumor medicine, in particular to an esophageal squamous cell carcinoma marker HES2 and its application in auxiliary diagnosis, treatment and prognosis judgment of esophageal squamous cell carcinoma. Background technique [0002] Esophageal cancer is a common malignant tumor. According to the 2012 global cancer statistics, there are an estimated 455,800 new cases of esophageal cancer worldwide. my country is one of the regions with a high incidence of esophageal cancer in the world, accounting for about 50% of the world's incidence, with an average of 150,000 deaths per year. [0003] The incidence and mortality of esophageal cancer vary greatly from country to country, and about 300,000 people die from esophageal cancer every year in the world. my country has become the country with the highest mortality rate of esophageal cancer in the world. About half of the cases diagnosed with esophage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/158
Inventor 宋立兵李隽林楚勇叶丽平李悦
Owner SUN YAT SEN UNIV CANCER CENT
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