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Application of loranoside C as Beclin1 activator in preparation of anti-hepatoma drugs

A technology of lanatoside C and an activator, which is applied in the field of biomedicine and can solve unclear problems

Pending Publication Date: 2022-04-05
广西皓智科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is still unclear whether lanatoside C can effectively activate Beclin1 and inhibit the proliferation of liver cancer Huh-7 cells

Method used

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  • Application of loranoside C as Beclin1 activator in preparation of anti-hepatoma drugs
  • Application of loranoside C as Beclin1 activator in preparation of anti-hepatoma drugs
  • Application of loranoside C as Beclin1 activator in preparation of anti-hepatoma drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, drug IC50 experiment

[0030] (1) After routinely digesting the cells with trypsin at room temperature, blow them into a uniformly distributed single-cell suspension with DMEM medium containing FBS, inoculate them in a 96-well plate at a density of 4,000 cells per well, and continue culturing at 37°C, 5%CO 2 in the incubator;

[0031] (2) After 24 hours of adherent cell growth, lanatoside C was added to make the final concentration 0.039, 0.078, 0.156, 0.312, 0.625, 1.25, 2.5, 5, 10, 20 μg / mL, and the control group was given an equal volume of DMSO ( That is, the DMSO dosage of 20 μg / mL lanatoside C was dissolved), and the cells were cultured for 1 day;

[0032] (3) Discard the cell culture medium, add 10 μL of CCK-8 solution to each well, and continue culturing for 2 h;

[0033] (4) Use a microplate reader to measure the absorbance at 450nm, compare the effects of different doses on cell proliferation inhibition, and calculate the cell proliferation in...

Embodiment 2

[0035] Example 2. Cell proliferation assay (CCK-8) detects the inhibitory effect of different concentrations of lanatoside C on the proliferation of liver cancer Huh-7 cells at different times

[0036] (1) After routinely digesting the cells with trypsin at room temperature, blow them into a uniformly distributed single-cell suspension with DMEM medium containing FBS, inoculate them in a 96-well plate at a density of 2,000 cells per well, and continue culturing at 37°C. 5%CO 2 in the incubator;

[0037] (2) After 24 hours of adherent cell growth, lanatoside C was added to make the final concentrations 0.078, 0.156, 0.312, 0.625, and 1.25 μg / mL, and the control group was given an equal volume of DMSO (that is, 1.25 μg / mL lanatoside C was dissolved with DMSO dosage of Glycoside C), culture cells for 2h, 1d, 2d, 3d, 4d, 5d;

[0038] (3) Discard the cell culture medium, add 10uL of CCK-8 solution to each well, and continue culturing for 2h;

[0039] (4) Use a microplate reader ...

Embodiment 3

[0041] Example 3. Cell clone formation experiment to detect the effect of lanatoside C on the clone formation of liver cancer Huh-7 cells

[0042] (1) After routinely digesting the cells with trypsin at room temperature, blow them into a uniformly distributed single-cell suspension with DMEM medium containing FBS, inoculate them in a 6-well plate at a density of 2,000 cells per well, and continue culturing at 37°C. 5%CO 2 in the incubator;

[0043](2) After culturing for 9 days, lanatoside C was added to intervene, so that the final concentration was 0.625 μg / mL, and the control group was given an equal volume of DMSO (that is, the cells were treated with DMSO dose of 0.625 μg / mL lanatoside C) , continue to cultivate for 7 days;

[0044] (3) Discard the culture medium in the upper layer, and wash the plate twice with PBS solution;

[0045] (4) Fix the cells with 4% paraformaldehyde for 15 minutes, discard the paraformaldehyde solution;

[0046] (5) Dyeing with crystal viol...

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Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to application of erioside C serving as an activator of Beclin1 in preparation of anti-liver cancer drugs. By up-regulating expression of Beclin1 protein, the chaenoside C promotes conversion from LC3I to LC3II and promotes formation of cell autophagosome, so that proliferation of liver cancer Huh-7 cells is inhibited. It is found in tests that the erioside C has an inhibiting effect on liver cancer Huh-7 cells, shows good time and concentration dependence, provides a new candidate drug for treating liver cancer by targeting Beclin1, and has important clinical significance.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of lanatoside C as an activator of Beclin1 in the preparation of anti-liver cancer drugs. Background technique [0002] Autophagy is a highly conserved lysosome-dependent pathway for the catabolism of intracellular substances in eukaryotic cells, through which intracellular components are recycled or energy is generated to maintain homeostasis. The autophagy process begins with the generation of a double-layer isolation membrane at the site of autophagosome formation, then the membrane edge elongates and closes to form an autophagosome, and finally the autophagosome fuses with a lysosome containing a variety of acid hydrolases to form Autophagy of lysosomes, thereby degrading the contents encapsulated in the inner membrane. Among them, the ubiquitin-like complex Atg12-Atg5-Atg16 formed in the cell performs the function of ubiquitin ligase, and this...

Claims

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Application Information

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IPC IPC(8): A61K31/7048A61P35/00
Inventor 谭宁吴丹李浩余想远廖洪涛薛宇周程杨凤娟
Owner 广西皓智科技有限公司
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