Application of composition of TRIM11 inhibitor and metformin in treatment of hepatocellular carcinoma

1. The technology of TRIM11 and metformin, applied in the field of biomedicine, can solve the problems of increased expression level, poor clinical prognosis, unclear mechanism of cell survival and tumor development, etc., to reduce blood sugar, reduce cell viability, and improve therapeutic effect Effect

Pending Publication Date: 2022-01-11
SHENZHEN INST OF ADVANCED TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the expression of TRIM11 was significantly upregulated in a variety of tumor tissues, and elevated TRIM11 expression levels were associated with poorer clinical prognosis
However, how TRIM11 regulates cell survival and tumor development under metabolic stress, especially in HCC, remains unclear

Method used

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  • Application of composition of TRIM11 inhibitor and metformin in treatment of hepatocellular carcinoma
  • Application of composition of TRIM11 inhibitor and metformin in treatment of hepatocellular carcinoma
  • Application of composition of TRIM11 inhibitor and metformin in treatment of hepatocellular carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Establishment of liver cancer cell line Huh7 stably overexpressing Flag-TRIM11 or knocking down TRIM11

[0050] Preparation of Huh7 cell lines stably overexpressing or knocking down TRIM 11:

[0051] A tumor cell line stably overexpressing or knocking down TRIM11 was constructed using lentivirus, and then the stable cell line was obtained by screening with puromycin. A part of the obtained Huh7 cell line stably overexpressing or knocking down TRIM11 was collected for western blot detection, the results showed that TRIM11 was overexpressed / knockdown, so far the Huh7 cell line with stable overexpression or knockdown of TRIM11 was obtained.

[0052] 1. Establishment of Huh7 cell line stably knocking down TRIM11

[0053] 1. Packaging Lentivirus

[0054] (1) 293T plating, 10cm dish, 2-4*10^6 cells / dish, 6ml DMEM+10%FBS medium, and transfection when the cell density reaches 60%-80%.

[0055] (2) Three-plasmid system transfected cells: pCMV-DR8.91: pCMV-VSVG: pLK...

Embodiment 2

[0094] 1. Use medium lacking glucose (Gibco TM Product No. 11966025, DMEM, without glucose, containing phenol red and glutamine, without glucose, sodium pyruvate and HEPES) was used to treat Huh7 cells and HepG2 cells according to certain time points (0h, 6h, 12h, 18h). Huh7 cells were collected to extract RNA. Genomic DNA was removed and reverse transcription was performed to obtain cDNA. Afterwards, qPCR was carried out, and the Ct value in the obtained result was divided into 2 -△△Ct The formula is used to calculate the relative expression level of mRNA of each gene. Western blot was performed on the collected Huh7 cells and HepG2 cells to detect the expression level of TRIM11 protein in the cells.

[0095] 2. Treat Huh7 cells with complete medium (0mM, 2.5mM, 5mM, 10mM) containing different concentrations of metformin (Hyclone product number SH30243.01, DMEM high glucose medium, containing glutamine and sodium pyruvate). After 12 hours, the cells were collected to extr...

Embodiment 3

[0102] 1. Stable knockdown control (shNC, PLKO.1) or TRIM11 (shTRIM11-1, -2) in Huh7 cells, using normal medium (Hyclone product number SH30243.01, DMEM high glucose medium, containing glutamine and sodium pyruvate) and glucose-deficient medium (Gibco TM Product No. 11966025, DMEM, without glucose, containing phenol red and glutamine, without glucose, sodium pyruvate and HEPES) for 6h and 12h, using the MTT method to measure the optical density (OD) at 490nm to measure cell viability.

[0103] 2. Stable overexpression of control (Vector, TRPE-GFP-mCherry) or TRIM11 in Huh7 cells, using normal medium (Hyclone product number SH30243.01, DMEM high glucose medium, containing glutamine and sodium pyruvate) and lack of Glucose medium (Gibco TM Product No. 11966025, DMEM, without glucose, containing phenol red and glutamine, without glucose, sodium pyruvate and HEPES) for 6h and 12h, using the MTT method to measure the optical density (OD) at 490nm to measure cell viability.

[010...

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Abstract

The invention discloses application of a composition of a TRIM11 inhibitor and metformin in preparation of a medicine for preventing or treating hepatocellular carcinoma, and particularly discloses application of a composition of a TRIM11 gene expression inhibitor and / or a TRIM11 protein activity inhibitor and metformin in preparation of a medicine for preventing or treating hepatocellular carcinoma. According to the invention, the TRIM11 expression is knocked down through the TRIM11 gene expression inhibitor or the TRIM11 protein activity inhibitor; the hepatocellular carcinoma is treated in cooperation with the metformin; therefore, the limitation of the current treatment of the hepatocellular carcinoma through the metformin treatment and the TRIM11 gene silencing expression is solved; and the synergistic cooperation of the metformin treatment and the TRIM11 inhibitor treatment provides the good anti-tumor effect.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to the application of a composition of a TRIM11 inhibitor and metformin in the preparation of a drug for preventing or treating hepatocellular carcinoma, more specifically to the combination of a TRIM11 gene expression inhibitor and / or a TRIM11 protein activity inhibitor and metformin Application of the composition in the preparation of drugs for preventing or treating hepatocellular carcinoma. Background technique [0002] Hepatocellular carcinoma (HCC) is the most common malignant tumor in the world, and its death rate ranks second among all cancers. The clinical prognosis of hepatocellular carcinoma is poor, mainly due to its high tumor cell proliferation potential, vascular invasion ability, tumor metastasis ability and postoperative recurrence. Although novel treatment methods such as targeted therapy are developing rapidly, due to the limited understanding of the occurrence...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/06A61K31/155A61K31/7088A61P1/16A61P35/00
CPCA61K45/06A61K31/155A61K31/7088A61P1/16A61P35/00A61K2300/00
Inventor 陈亮朱燕
Owner SHENZHEN INST OF ADVANCED TECH
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