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Hepatoma marker OXCT1 and application thereof to hepatoma diagnosis, treatment and prognosis

A liver cancer prognosis and marker technology, applied in treatment and prognosis, OXCT1 in the field of liver cancer diagnosis and potential liver cancer markers can solve problems such as unclear expression and mechanism of action, and inability to use ketone bodies

Active Publication Date: 2017-10-24
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the lack of the key rate-limiting enzyme OXCT1 (protein name SCOT) for utilizing ketone bodies in the liver, the liver itself cannot utilize ketone bodies
At present, the expression and mechanism of OXCT1 in human liver cancer cells are still unclear.

Method used

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  • Hepatoma marker OXCT1 and application thereof to hepatoma diagnosis, treatment and prognosis
  • Hepatoma marker OXCT1 and application thereof to hepatoma diagnosis, treatment and prognosis
  • Hepatoma marker OXCT1 and application thereof to hepatoma diagnosis, treatment and prognosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: The expression of OXCT1 gene / protein is up-regulated after being stimulated by serum starvation in liver cancer, and the expression is up-regulated in clinical samples of liver cancer

[0030] 1. Western blot to detect the protein expression of gene OXCT1 in different tissues of mice

[0031] Methods: The tissue samples of brain, heart, liver, kidney, lung and stomach were obtained by dissecting nude mice, and each tissue was broken into tissue homogenate with a tissue breaker, and then passed through cell lysate (50mM Tris-Cl, pH8.0, 150mMNaCl, 5mM EDTA, 0.1% SDS, 1% NP-40, protease cocktails) to disrupt the cells of each component, and the cell content (including protein) was incorporated into the cell lysate, and the protein in the cell lysate was quantified under the same conditions, and finally the same Quantitative samples of each component were detected and analyzed by Western bolt, and the specific steps were as follows: after separating the protein b...

Embodiment 2

[0050] Example 2: Construction of a cell line stably knocking down OXCT1 and a cell line stably overexpressing OXCT1.

[0051] 1. Construction of a cell line that stably knocks down OXCT1 mRNA and protein

[0052] (1) Construct the expression plasmid of shOXCT1

[0053] Prepare the hairpin structure (hairpin structure) shOXCT1 that knocks down the expression of OXCT1 gene. After the hairpin structure (hairpin structure) is integrated into the cell genome, it can express siRNA. siRNA and RISC inhibit mRNA translation or cleave mRNA, thereby reducing the target gene in the cell level of expression. The above-mentioned shOXCT1 can be synthesized by relevant biological companies, and the shOXCT1 used in the present invention includes a broad-spectrum shRNA that knocks down the expression of the OXCT1 gene and an shRNA that knocks down the expression of the endogenous OXCT1 gene. The shRNA for broad-spectrum knockdown of OXCT1 was purchased from the shRNA library of sigma company...

Embodiment 3

[0065] Example 3: Functional research of OXCT1 protein in liver cancer cells

[0066] 1. Detection of tumor cell proliferation

[0067] (1) In vitro experiment: cell growth experiment

[0068] The number of cells in different groups was counted by bromophenol blue staining, and the changes in the growth rate of liver cancer cells with knockdown of OXCT1 gene / protein and exogenous overexpression of OXCT1 gene / protein were detected. The brief description is as follows: Cells in the experimental group (HepG2 / Hep3B-shOXCT1 and HepG2 / Hep3B-OXCT1) or cells in the control group (HepG2 / Hep3B-NTC Control and HepG2-EV Control) were divided into 5×10 4 Each well was inoculated in a 24-well culture plate. After 24 hours, the 10% fetal bovine serum (Gibco BRL) in DMEM medium (DMEM; Gibco BRL) was replaced with 0% or 1%, and the corresponding wells were replaced every 24 hours. The cells in the medium were digested and blown into single cells, diluted to a certain multiple with bromopheno...

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Abstract

The invention provides a hepatoma marker OXCT1 and application thereof to hepatoma diagnosis, treatment and prognosis. The inventor discovers that the expression of OXCT1 in the hepatoma tumor in the body is obviously higher than the expression in the normal liver tissue; the cell proliferation can be obviously reduced through endogenous OXCT1 inhibition; in addition, the survival rate of hepatoma patients with higher OXCT1 expression is lower, i.e., the over expression of the OXCT1 definitely indicates the relatively poor survival prognosis. On the basis, the OXCT1 can be the tumor auxiliary diagnosis marker, and can be effective target points for tumor treatment, and can also be survival prognosis indexes of the hepatoma patients.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to a potential liver cancer marker OXCT1. The OXCT1 gene plays a role in the process of causing liver cancer, and relates to the application of OXCT1 in the diagnosis, treatment and prognosis of liver cancer. Background technique [0002] Malignant tumors seriously endanger human health and have surpassed cardiovascular disease as the leading cause of death. According to the 2012 global cancer statistics report: 695,000 people died of liver cancer in the world that year, ranking third in the death rate of all cancers. What's more serious is that China is the country with the highest incidence of liver cancer and the highest number of deaths in the world. The number of liver cancer patients in my country accounts for about 52.7% of the global liver cancer patients. And the number of patients with liver cancer in my country is increasing rapidly every year. It is estimated that ...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N15/54C12Q1/68G01N33/574G01N33/573A61K48/00A61K31/7088A61K39/395A61P35/00
CPCA61K31/7088A61K39/395C12N9/13C12Q1/6886C12Q2600/118C12Y208/03005G01N33/573G01N33/57438G01N2333/91194
Inventor 张华凤高平黄的李婷婷
Owner UNIV OF SCI & TECH OF CHINA
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