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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: 2012-12-12
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
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  • 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 ability

[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 Mechani sm of Ammonia-induced Astrocyte Swelling. The Journal of Biological Chemi stry. 2008, 283(49): 33875.

[0033] Perform the following operations on MHCC97L, MHCC97H and HCCLM3 respectively:

[0034] 1. Press the cell 2×10 5 Cells / ml were inoculated on a six-well culture plate, each well was inoculated with 2ml cell suspension, placed at 37℃, 5% CO 2 Culture in an incubator; after the cells have grown to 80-90% confluence, aspirate the liquid and add DMEM high-sugar medium at 37℃, 5% CO 2 Continue to incubate for 30 min in the incubator.

[0035] 2. Aspirate the medium...

Embodiment 2

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

[0040] 1. Fluorescence spectrophotometer detection

[0041] 1. Liver cancer cell line MHCC97H

[0042] (1) Press 2×10 of the liver cancer cell line MHCC97H 5 Cells / ml were inoculated on a six-well culture plate, each well was inoculated with 2ml cell suspension, placed at 37℃, 5% CO 2 Culture in an incubator.

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

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

[0045] The second group (MHCC97H+BUM): DMEM high sugar medium containing 50μM bumetanide was added;

[0046] At 37℃, 5% CO 2 Continue to incubate for 30 min in the incubator.

[0047] (3) Aspirate the medium and add DMEM high glucose medium containing 0.05% (volume percentage) pluronic acid and 10μM sodium binding benzofuran isophthalate acetoxy methyl esters-AM (SBFI) at 37℃, 5% CO 2 Incubate for 2h in an incubator p...

Embodiment 3

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

[0080] 1. Liver cancer cell line MHCC97H

[0081] 1. Take the hepatocarcinoma cell line MHCC97H in the logarithmic growth phase (70%-80% confluence) and digest the cell suspension with 0.25% pancreatin (Hyclone).

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

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

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

[0085] Place at 37℃, 5% CO 2 Incubate in an incubator for 12 hours.

[0086] 3. Use the CCK-8 kit to measure the absorbance of each well at the OD450nm of the microplate reader at 0 days, 1 day, 2 days, 3 da...

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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 use of bumetanide, in particular to the application of bumetanide in inhibiting the metastasis of liver cancer cells. Background technique [0002] More than 90% of primary liver cancers are hepatocellular carcinoma (HCC). HCC is a very common malignant tumor with a very high fatality rate. About 650,000 people die of HCC every year, and its incidence is increasing year by year. the trend of. HCC occurs mostly in developing countries such as Southeast Asia and Africa, and more than 50% of them occur in China, ranking second in cancer mortality in my country. 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. Clinical surgical resection of liver tumors and liver transplantation are considered to be the best means to obtain a radical cure for HCC, but only 10-20% of HCC patients are suitable for surgical treatment;...

Claims

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Application Information

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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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