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Application of mir‑455‑3p in the diagnosis, treatment and prognosis of esophageal squamous cell carcinoma

A technology for esophageal squamous cell carcinoma, applied in gene therapy, medical preparations containing active ingredients, biochemical equipment and methods, etc., can solve problems such as difficulty in killing tumor stem cells

Active Publication Date: 2017-06-30
SUN YAT SEN UNIV CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional drugs are difficult to kill cancer stem cells

Method used

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  • Application of mir‑455‑3p in the diagnosis, treatment and prognosis of esophageal squamous cell carcinoma
  • Application of mir‑455‑3p in the diagnosis, treatment and prognosis of esophageal squamous cell carcinoma
  • Application of mir‑455‑3p in the diagnosis, treatment and prognosis of esophageal squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 miR-455-3p is upregulated in ESCC samples

[0041] (1) TCGA database analysis

[0042] Methods: Using SPSS statistical analysis method, the expression level of miR-455-3p and its relationship with survival rate were analyzed in 186 ESCC patients and 13 ESCC paracancerous tissues in The Cancer Genome Altas Project (TCGA). All statistical analyzes were processed with SPSS17.0 statistical software. Values ​​are expressed as mean ± SD, P <0.05 was considered a statistically significant difference.

[0043] The analysis results showed that the expression of miR-455-3p in ESCC tumor tissue was significantly higher than that in paracancerous tissue samples ( P figure 1 -A); define the expression of miR-455-3p in ESCC tumor tissue exceeding the median expression, called miR-455-3p high expression; ESCC tumor tissue miR-455-3p expression lower than the median expression, called miR-455-3p low expression, from figure 1 In -B, it can be seen that the high expressi...

Embodiment 2

[0053] Example 2 Construction of ESCC tumor cell lines resistant and sensitive to chemotherapeutic drugs

[0054] (1) Establishment of drug-resistant strains and non-drug-resistant strains

[0055] Methods: The tumor cells of ESCC patients were subcutaneously inoculated into NOD / SCID mice (4–5 weeks old, 18-20g), and one week later, the mice were intraperitoneally injected with PBS buffer or 5mg / kg cis-diamine Chloroplatinum (CDDP), twice a week for 4 weeks. Afterwards, the ESCC tumor cells in the mice were isolated and cultured on a plate, and the above steps were repeated 3 times. The mouse tumor cells after the fourth inoculation were isolated and cultured on a plate. The ESCC tumor cells obtained in the CDDP group were CDDP-treated ESCC strains (hereinafter referred to as EC-CR), and those in the PBS group were non-CDDP-treated ESCC strains (hereinafter referred to as EC-CR). EC-NT) (see 2-A). EC-CR and EC-NT cells were treated with chemotherapeutic drugs cis-diamminodi...

Embodiment 3

[0074] Example 3 Construction of miR-455-3p stable high expression and inhibited expression cells

[0075] (1) microRNA associated with ESCC tumor stem cells

[0076] Using microRNA expression profile analysis to detect the expression of microRNA in EC-CR and EC-NT cells, it was found that miR-455-3p was highly expressed in EC-CR and EC-NT cells, and the expression of miR-455-3p in EC-CR The expression level is higher than that of EC-NT (see image 3 -A).

[0077] (2) Construction of stable cells with high expression of miR-455-3p

[0078] ESCC cells EC-NT were cultured in DMEM medium (DMEM; Gibco) supplemented with 10% fetal bovine serum (Gibco) in a carbon dioxide incubator at 37°C.

[0079] 1) Construct the overexpression plasmid of miR-455-3p

[0080] The full-length mRNA sequence of miR-455-3p, UCCCUGGCGUGAGGGUAUGUGCCUUUGGACUACAUCGUGGAAGCCAGCACCAUGCAGUCCAUGGGCAUAUACACUUGCCUCAAGGCCUAUGUCAUC (SEQ ID NO: 5), was amplified by PCR.

[0081] The primer sequences are as fol...

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Abstract

The invention discloses applications of miR-455-3p in the diagnosis, treatment, and prognosis of esophageal squamous cancer. The inventor finds that the obviously low survival rate of ESCC patients is related with the high expression of miR-455-3p in ESCC patients. The in-vitro and in-vivo experiment results show that miR-455-3p is capable of promoting the malignant progression of ESCC, by inhibiting the expression of miR-455-3p, the ESCC stem cell characteristics can be inhibited, and the sensitivity of ESCC stem cells to chemotherapy drugs is enhanced; so an expression inhibitor of miR-455-3p can inhibit the tumor stem cells, and is capable of inhibiting the tumor stem cell characteristics and enhancing the sensitivity of the cancer stem cells to chemotherapy drugs. The invention finds the expression of miR-455-3p in cancer patients for the first time, and the expression can be used as auxiliary diagnosis and / or prognosis of ESCC. The invention provides a novel diagnosis method, a novel treatment method, and a pharmaceutical drug screening platform for tumor diseases.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a new microRNA and the application of Antagomir, in particular to the application of miR-455-3p and Antagomir-455-3p. Background technique [0002] Esophageal squamous cell carcinoma (ESCC) is one of the most common malignant tumors in the world, and its death rate ranks sixth. About 90% of esophageal cancer in my country is squamous cell carcinoma, and it has significant regional characteristics. The incidence of esophageal squamous cell carcinoma in high-incidence areas is 500 times that in low-incidence areas. Smoking and alcohol history, nutritional deficiencies, hot food, and intake of carcinogens are all associated with esophageal squamous cell carcinoma. Surgery, radiotherapy and chemotherapy are important treatments for esophageal squamous cell carcinoma, but the 5-year survival rate is only about 10%. Drug resistance is a difficult problem in the treatment of esophageal squam...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68A61K48/00A61K31/7088A61P35/00
CPCA61K31/7088C12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 宋立兵李隽刘爱斌曹丽雪朱金容吴阁艳吴淑
Owner SUN YAT SEN UNIV CANCER CENT
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