Application of bumetanide in inhibition of hepatoma cell transfer

A technology of liver cancer cells and cells, which is applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problem of renal tubule ineffectiveness and achieve the effect of inhibiting proliferation

Active Publication Date: 2014-09-24
BEIJING PROTEOME RES CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Bumetanide is a diuretic that mainly inhibits the active reabsorption of NaCl in the thick-walled segment of the ascending branch of the loop of Henle of the renal tubule, and reabsorbs NaCl in the proximal tubule. + Also inhibits, but has no effect on distal tubules

Method used

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  • Application of bumetanide in inhibition of hepatoma cell transfer
  • Application of bumetanide in inhibition of hepatoma cell transfer
  • Application of bumetanide in inhibition of hepatoma cell transfer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Comparison of the expression of NKCC1 activity in liver cancer cell lines with different metastatic abilities

[0032] In this example, the expression of NKCC1 activity in different liver cancer cell lines was compared by detecting the sodium ion content in the protein solution. For the relevant mechanism, see the literature AR.Jayakumar, ML.Liu, M.Moriyama.et al.Na-K-Cl Cotransporter-1 in the Mechanism of Ammonia-induced Astrocyte Swelling. The Journal of Biological Chemistry. 2008, 283(49): 33875.

[0033] MHCC97L, MHCC97H and HCCLM3 are respectively operated as follows:

[0034] 1. Divide cells into 2×10 5 Inoculate in a six-well culture plate at a density of cells / ml, inoculate 2ml of cell suspension in each well, and place at 37°C, 5% CO 2 Cultivate in an incubator; after the cells grow to 80-90% confluence, discard the liquid and add DMEM high glucose culture at 37°C, 5% CO 2 Continue culturing for 30 min in the incubator.

[0035] 2. Discard the cu...

Embodiment 2

[0039] Example 2. Detection of the effect of bumetanide on the activity of NKCC1 in liver cancer cells

[0040] 1. Fluorescence spectrophotometer detection

[0041] 1. Liver cancer cell line MHCC97H

[0042] (1) Liver cancer cell line MHCC97H was divided into 2×10 5 Inoculate in a six-well culture plate at a density of cells / ml, inoculate 2ml of cell suspension in each well, and place at 37°C, 5% CO 2 Incubator cultivation.

[0043] (2) After the cells grow to 80-90% confluence, discard the liquid and divide into two groups:

[0044] The first group (MHCC97H): add DMEM high glucose medium;

[0045] The second group (MHCC97H+BUM): add DMEM high-glucose medium containing 50 μM bumetanide;

[0046] At 37°C, 5% CO 2 Continue culturing for 30 min in the incubator.

[0047] (3) Discard the medium, add DMEM containing 0.05% (volume percent) pluronic acid and 10 μM sodium binding benzofuran isophthalate acetoxy methyl esters-AM (SBFI) to culture at 37°C, 5% CO 2 Incubate in th...

Embodiment 3

[0079] Example 3. Effect of bumetanide on proliferation ability of liver cancer cell lines MHCC97H and Huh7 cells in vitro

[0080] 1. Liver cancer cell line MHCC97H

[0081] 1. The liver cancer cell line MHCC97H in the logarithmic growth phase (70%-80% confluence) was digested with 0.25% trypsin (Hyclone Company) to form a cell suspension.

[0082] 2. Divide the wells in the 96-well plate into two groups:

[0083] The first group (MHCC97H): add DMEM high-glucose medium containing 10% (volume percentage) FBS, then inoculate cell suspension, and the cell concentration is 1000 cells / μl;

[0084] The second group (MHCC97H+BUM): Add DMEM high-glucose medium containing 10% (volume percentage) FBS and 50 μM bumetanide, and then inoculate the cell suspension with a cell concentration of 1000 cells / μl;

[0085] Placed at 37°C, 5% CO 2 Culture in the incubator for 12 hours.

[0086] 3. Use the CCK-8 kit to measure the absorbance of each well at the OD450nm of a microplate reader on...

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Abstract

The invention discloses application of bumetanide in inhibition of hepatoma cell transfer. Bumetanide can be used for preparing a medicine which is used for inhibiting hepatoma cell multiplication and / or hepatoma cell attack and / or hepatoma cell transfer, can be used for preparing a medicine which is used for curing hepatoma cells and can be used for preparing a product which is used for reducing phosphorylation NKCC1 protein content in hepatoma cells. The bumetanide can be used for reducing the phosphorylation NKCC1 protein content in hepatoma cells, so that hepatoma cell multiplication, attack and transfer are inhibited. The application has significant value for hepatoma cell cure.

Description

technical field [0001] The invention relates to a new application of bumetanide, in particular to the application of bumetanide in inhibiting liver cancer cell metastasis. Background technique [0002] More than 90% of primary liver cancer is hepatocellular carcinoma (HCC). HCC is a very common malignant tumor with a very high lethality rate. About 650,000 people die from HCC every year, and its incidence is increasing year by year the trend of. HCC mostly occurs in developing countries such as Southeast Asia and Africa, and more than 50% of them occur in China, ranking second in my country's cancer mortality rate. In addition, the incidence of HCC in developed countries is also increasing year by year. Early diagnosis of HCC has always been a bottleneck in clinical treatment. Clinically, surgical resection of liver tumors and liver transplantation are considered to be the best means to achieve a radical cure for HCC, but only 10-20% of HCC patients are suitable for surgic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/196A61P35/04A61P35/00C12N5/09
Inventor 姜颖贺福初周亚亚孙薇林巍然魏汉东
Owner BEIJING PROTEOME RES CENT
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