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Human lung cancer paraquat-resistant cell strain

A technology of paraquat and cell lines, applied in the field of cell engineering, to achieve obvious anti-PQ toxicity characteristics and improve the induction success rate

Active Publication Date: 2016-03-02
THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Through searching, it is found that there is no literature report on the establishment of human lung cancer drug-resistant paraquat cell line by using the human lung cancer cell line A549 as the induction object at present.

Method used

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  • Human lung cancer paraquat-resistant cell strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Induction and establishment of paraquat-resistant human lung cancer cell lines

[0028] The human lung cancer-resistant paraquat cell line A549 / PQ was established according to the following steps: (1) A549 cells were routinely cultured in 1640 medium containing 10% fetal bovine serum at 37°C, 5% CO 2 . The cells in the logarithmic growth phase were used for the experiment each time. (2) Use the PQ induction method of increasing concentration and periodic exposure to establish cell resistant strains: the concentration of PQ in the culture medium starts from 100 μmol / L, and the solution is changed after adding the drug for 24 hours to remove the drug, and the cells will re-grow to the logarithmic phase Then add the same concentration of PQ to stimulate again, repeat 3 times. Then extend the exposure time to 48h (ie 100μmol / L exposure for 48h), change the medium to remove the drug, and after the cells re-grow to the logarithmic phase, give the same drug for the sam...

Embodiment 2

[0030] Example 2 Morphological observation of human lung cancer-resistant paraquat cell line A549 / PQ and human lung cancer A549 cells

[0031] Take the human lung cancer cell lines A549 and A549 / PQ cells in the logarithmic growth phase, after routine digestion and passage, use a Leica optical microscope to observe the cell morphology after the cells are fully attached 24 hours, and take pictures with a Nikon camera. The results are as follows figure 1 Shown.

Embodiment 3

[0032] Example 3 The cell counting method draws the growth curve of the human lung cancer-resistant paraquat cell line A549 / PQ and the human lung cancer cell line A549 and determines the doubling time

[0033] Take 50,000 parental A549 cells and human lung cancer paraquat-resistant cell line A549 / PQ cells that are in the middle logarithmic growth phase and have good growth conditions to prepare cell suspensions and inoculate them into culture flasks. Each cell is inoculated into 21 flasks. Start cell counting after 24 hours, count for seven consecutive days, count three bottles of cells every day, and draw cell growth curves based on the counting results. Then calculate the doubling time (DT) according to the formula, DT=t×lg2 / (lgNt-lgN0), t is the cell culture time, N0 is the average number of cells measured 24h after inoculation, and Nt is the average number of cells after culture th Value, the result is figure 2 with 3 Shown.

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Abstract

The invention provides a human lung cancer paraquat-resistant cell strain. A human lung cancer cell strain A549 is used as an induction object; an in-vitro induction method of periodic exposure and progressive increase of paraquat concentration is adopted; and a paraquat-resistant cell strain A549 / PQ is built. The cell strain is collected in CGMCC (China General Microbiological Culture Collection Center) on June 16, 2015; and the collection number is CGMCC NO.10892. The cell strain has the typical paraquat-resistant characteristic; by using the biological characteristic, a study model can be provided for further studying a paraquat toxicity reversion path, extracting a paraquat poisoning evaluation marker, exploring a novel anti-oxidization path and the like; and high scientific research and production and application values are realized.

Description

Technical field [0001] The invention belongs to the technical field of cell engineering, and relates to a human cell line, in particular to a human lung cancer-resistant paraquat cell line and its establishment method and application. Background technique [0002] Paraquat (PQ) is a highly effective herbicide widely used worldwide. It is highly toxic to the human body. It can cause poisoning when absorbed through the digestive tract, skin, respiratory tract, and vein, and there is no clinically effective treatment. Measures, the patient mortality rate is as high as 40 to 80%. The production and consumption of PQ in my country rank first in the world. In recent years, the incidence of PQ poisoning in China has multiplied, and the prevention and treatment situation is very severe. Although my country began to gradually ban the production and sales of PQ liquid in June 2015, other dosage forms will continue to be produced and used. Therefore, paraquat poisoning is still an import...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
Inventor 洪广亮卢中秋刘佳明谭佳平丁颖威赵光举
Owner THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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