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Application of cell cycle blocker KR in human lung cancer cells

A human lung cancer cell line, cell cycle technology, applied in the field of biochemistry and molecular biology, can solve problems such as toxic and side effects

Inactive Publication Date: 2017-08-29
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conventional drugs clinically used to treat lung cancer, although they play an important role in the treatment, have obvious toxic and side effects

Method used

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  • Application of cell cycle blocker KR in human lung cancer cells
  • Application of cell cycle blocker KR in human lung cancer cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] In vitro proliferation inhibitory activity of KR on human lung cancer cell line A549: the resuscitated human lung cancer cell line A549 was cultured in sterile RPMI1640 containing 10% FBS inactivated fetal bovine serum, 1% penicillin / 1% streptomycin (double antibody) In the culture medium, put in the suspension cell flask, at 37°C, 5% CO 2 and cultured in an incubator under saturated humidity, and the cells grow to about 70%-80% for one passage. Continue culturing to obtain a human lung cancer cell line 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 of RPMI 1640 culture medium in each well. According to the experimental requirements, 10 μl of 120 μM, 200 μM, 100 μM, 20 μM, 10 μM, and 2 μM KR were added to the treatment group, and incubated in an incubator for 2 days. 4 replicate wells each. Use MTT kit for detection, add 20 μl MTT solution to each well of cell...

Embodiment 2

[0022] KR induces apoptosis of A549 cells: Digest the above-mentioned cultured cells in the logarithmic growth phase with trypsin, count the cells, take an appropriate amount of cells to dilute, and the diluted cell density is 1×10 5 ~1.5×10 5 cells / ml, blow and mix well, inoculate the cells in a 6-well plate, take 2ml of human lung cancer cell line A549 from each well, and after 12h, after the cells adhere to the wall, set up a blank control group and a 10μΜKR group, and add 200μl of 100μΜKR to the 10μΜKR group (Working concentration is 10μΜKR), after mixing gently, place in a concentration of 5% CO 2 , incubate in an incubator with a relative humidity of 90% and a temperature of 37°C for 36 hours, divide into two samples with a 15ml centrifuge tube, fix the cells according to the instructions of the cell cycle and apoptosis detection kit, and prepare propidium iodide staining buffer. After 12 hours, add 10ml PBS to each sample to wash twice, resuspend the cells, add 500μl p...

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Abstract

The invention provides application of cell cycle blocker KR in human lung cancer cells, and belongs to the technical field of biochemistry and molecular biology. The application of the cell cycle inhibitor KR in the human lung cancer cells comprises the following steps: (1) cell recovery, (2) cell culture, (3) drug preparation, (4) cell treatment, (5) detection of cell proliferation inhibition rate and (6) detection of cell cycle. The data of the method is based on the fact that the KR can upregulate CDKN1A to cause ATP depletion and cause the cell cycle to be retarded at G0 / G1 phase, and has very significant in vitro proliferation inhibitory activity.

Description

technical field [0001] The invention relates to the anti-tumor effect of cytokinin kinetin riboside (KR) as a cell cycle blocker on lung cancer cell lines, and belongs to the technical field of biochemistry and molecular biology. Background technique [0002] Lung cancer is currently the malignant tumor with the highest morbidity and mortality worldwide. In the past 20 years, due to vigorous promotion of smoking cessation, the incidence of male lung cancer in Europe and the United States has begun to decline, but the incidence of female lung cancer has continued to rise. my country is a big country in the production and sales of cigarettes. Regardless of male or female, the incidence of lung cancer continues to rise, especially in females. Clinical studies have shown that the cure rate of carcinoma in situ is close to 100%, the 5-year survival rate of stage I lung cancer patients is 60%-90%, while the 5-year survival rate of IIIb and IV patients is only 5%-20%. Conventiona...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
CPCG01N33/5011G01N33/5044
Inventor 王黎程娇崔昌浩林凡琳张瀚文于冬丽邓营营
Owner DALIAN UNIV OF TECH
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