Application of apoptosis-inducing reagent in human breast cancer cell

A technology of human breast cancer cells and cells, applied in the fields of biochemistry and molecular biology, to achieve the effect of significant in vitro proliferation inhibitory activity

Inactive Publication Date: 2017-10-20
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on whether oTR exerts anti-tumor effect by inducing apoptosis

Method used

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  • Application of apoptosis-inducing reagent in human breast cancer cell
  • Application of apoptosis-inducing reagent in human breast cancer cell
  • Application of apoptosis-inducing reagent in human breast cancer cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] In vitro proliferation inhibitory activity of oTR on human breast cancer cells: the revived human breast cancer cells MCF-7 were cultured in a medium containing 10% FBS inactivated fetal bovine serum, 1% glutamine, 1% penicillin / 1% streptomycin ( Double antibody) sterile RPMI 1640 culture medium, placed in a cell culture flask, at 37 ° C, 5% CO 2 and cultured in an incubator under saturated humidity, and the cells grow to about 70% to 80%, and are passaged once. Continue culturing to obtain human breast cancer cells in an active growth state, and collect the cells. 5×10 that will be in logarithmic growth phase 3 Each cell was seeded in a 96-well plate containing 100 μl RPMI 1640 culture medium in each well. According to the experimental requirements, the treatment group was added with 0.1% DMSO, 0.3 μM, 1.5 μM, 15 μM, and 30 μM concentrations of oTR, and incubated in an incubator for 1 ~ 4 days, each with 4 replicate wells. Use the MTT kit for detection, add 20 μl MT...

Embodiment 2

[0023] oTR induces apoptosis of MCF-7 cells: After the actively growing MCF-7 cells were treated with 10 μM oTR for 48 hours, the staining buffer was prepared according to the instructions of the Annexin V-FITC apoptosis detection kit, and 500 μl of staining buffer was added to each sample Incubate at 37°C in the dark for 30 min, and finally wash once with PBS. After filtering with a 300-mesh sieve, it was detected with a BD FACSCalibur flow cytometer, and the apoptosis rate was calculated. The result is as image 3 as shown, image 3It can be seen that there are early and late apoptotic cell populations. Compared with the control group (1.87%), the early apoptosis of MCF-7 cells in the oTR treatment group (9.43%) was significantly increased ( image 3 ), and the late apoptosis control group (11.14%) was also less than the oTR treatment group (22.12%), indicating that under the same experimental conditions, oTR can induce apoptosis of MCF-7 cells.

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Abstract

The invention provides an application of an apoptosis-inducing reagent in a human breast cancer cell and in particular relates to an antitumor effect of cytokinin ortho-Topolin Riboside as a demethylation reagent on a human breast cancer cell strain and belongs to the technical fields of biochemistry and molecular biology. Data in the invention are established based on quite remarkable in-vitro proliferation inhibitory activity of oTR on 60 tumor cell lines of 9 tumour series of human, in particular on relatively high drug susceptibility of the human breast cancer cell strain.

Description

technical field [0001] The invention relates to the antitumor effect of cytokinin ortho-Topolin Riboside (oTR) as a demethylation agent on human breast cancer cell lines, and belongs to the technical field of biochemistry and molecular biology. Background technique [0002] Breast cancer is a gene mutation in breast glandular epithelial cells under the action of various carcinogenic factors, resulting in uncontrolled cell proliferation. As the biological behavior of cancer cells has changed, it presents disordered and unrestricted malignant proliferation. Its histological manifestation is that a large number of immature cancer cells proliferate indefinitely and crowd into groups in a disorderly manner, extruding and eroding to destroy the surrounding normal tissues and destroy the normal tissue structure of the breast. After the breast cells mutate, they lose the characteristics of normal cells. The tissue structure is disordered, and the cell connections are loose. Healin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7076A61P35/00
CPCA61K31/7076
Inventor 崔昌浩林凡琳王黎程娇张瀚文于冬丽邓营营
Owner DALIAN UNIV OF TECH
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