Establishing method and applications of cell model capable of simulating lung cancer occurrence mechanism

A cell model and mechanism technology, applied in the field of cell medicine, can solve the problems of complex pathogenesis of lung cancer and single inducing factor, etc.

A cell model and mechanism technology, applied in the field of cell medicine, can solve the problems of complex pathogenesis of lung cancer and single inducing factor, etc.

CN109294990AInactive Publication Date: 2019-02-01FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
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  • Establishing method and applications of cell model capable of simulating lung cancer occurrence mechanism
  • Establishing method and applications of cell model capable of simulating lung cancer occurrence mechanism
  • Establishing method and applications of cell model capable of simulating lung cancer occurrence mechanism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Conditional exploration and model construction of the effect of spherical SNPs on BPDE-infected BEAS-2B cells co-cultured with THP-1 cells

[0045] 1Exploration of optimal concentration

[0046] The medium used for culturing BEAS-2B and THP-1 cells is LHC-9 medium containing 10% FBS, and the medium is replaced every 2-3 days.

[0047] 1.1 Exploration steps

[0048] ⑴ Set the complete medium as the blank group, and the cells without any poison treatment as the control group, and set 3 parallel holes for each dose experimental group;

[0049] ⑵Adjust the concentration of BEAS-2B cells or THP-1 cells to 1×10 5 Pcs / ml, evenly seeded in 96-well plate, 200ul per well, after the cells grow adherently for 24 hours, discard the original culture medium;

[0050] ⑶ The concentrations of BPDE added are 0, 200nmol / ml, 400nmol / ml, 800nmol / ml and 1.2umol / ml; the concentrations of spherical SNPS are 0, 1.5625μg / ml, 3.125μg / ml, 6.25μg / ml, 12.5μg / ml and 25μg / ml;

[0051] ⑷ Discard the cu...

Embodiment 2

[0085] Example 2: Comparison of cytokine expression in serum of established cell model and actual lung cancer patients

[0086] 1 Cytokine detection in cell model

[0087] 1.1 Grouping

[0088] The experiment is divided into two groups: the control group is BEAS-2B cells + 800 nmol / L BPDE stimulation + THP-1; the experimental group is BEAS-2B cells + 800 nmol / L BPDE stimulation + THP-1 + 12.5 μg / ml spherical SNPs .

[0089] 1.2 Experimental steps

[0090] (1) Suspension chip experiment

[0091] The two groups of cells were cultured for 24 hours and 48 hours to detect the culture supernatant. According to the instruction of the suspension chip kit (Bio-Plex), the expression level of each cytokine was detected.

[0092] (2) Realtime-PCR experiment

[0093] The two groups of cells were cultured for 24 hours and 48 hours to detect the culture supernatant. RNA was extracted from the two groups of cells for RT-qPCR detection, and the BEAS-2B and THP-1 cells infected with BPDE and BPDE-SNPs we...

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Abstract

The invention relates to the field of cell medicine, particularly to an establishing method and applications of a cell model capable of simulating lung cancer occurrence mechanism, and provides an establishing method of a cell model capable of simulating lung cancer occurrence mechanism. The establishing method comprises: inducing the malignant transformation of BEAS-2B cells in a BEAS-2B cell andTHP-9 cell co-culture system by using BPDE and spherical SNPs as inducing agents. According to the present invention, the established cell model can highly simulate the lung cancer induction processhaving the same pathogenesis as Xuanwei lung cancer, and has application value in the study of lung cancer occurrence and the screening of lung cancer prevention and treatment drugs.

Description

Technical field [0001] The invention relates to the field of cell medicine, in particular to a method and application for establishing a cell model that simulates the mechanism of lung cancer. Background technique [0002] Xuanwei is located in southwest China and is a rich coal mine county in Qujing, Yunnan Province. Xuanwei County of 6,257 square kilometers has a population of 1.2 million. However, due to the high incidence of lung cancer, it has also received extensive attention from many medical researchers at home and abroad. A large amount of data shows that in 2004-2005, there were 91 deaths from lung cancer for every 100,000 people who died of lung cancer, which is much higher than the average international lung cancer mortality rate (31 deaths from lung cancer per 100,000 people). Earlier research data indicated that the main source of energy for heating and cooking rice in Xuanwei rural areas is bituminous coal produced locally in Xuanwei. However, almost no smoke ex...

Claims

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Application Information

Patent Timeline
01 Feb 2019
Publication
CN109294990A
IPC
C12N5/0786; C12N5/071; C12Q1/02; G01N33/68; G01N33/574
CPC
C12N5/0625; C12N5/0645; C12N5/0688; C12N2500/05; C12N2500/30; C12N2500/82; G01N33/5011; G01N33/57423
Inventors
单斌; 刘子杰