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Method for adjusting sensibility of lung adenocarcinoma cells to EGFR-TKIs by mediating progesterone by mPR alpha

A lung adenocarcinoma cell, sensitive technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problem of no related research on sensitivity

Active Publication Date: 2016-03-23
XIANGYA HOSPITAL CENT SOUTH UNIV
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  • Abstract
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Problems solved by technology

However, there is no relevant research in the field of mPRα mediating progesterone to regulate the sensitivity of lung adenocarcinoma cells to EGFR-TKIs

Method used

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  • Method for adjusting sensibility of lung adenocarcinoma cells to EGFR-TKIs by mediating progesterone by mPR alpha
  • Method for adjusting sensibility of lung adenocarcinoma cells to EGFR-TKIs by mediating progesterone by mPR alpha
  • Method for adjusting sensibility of lung adenocarcinoma cells to EGFR-TKIs by mediating progesterone by mPR alpha

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experiment example

[0121] 1.1 Materials

[0122] 1.1.1 Cells

[0123] Lung adenocarcinoma cell line A549 was purchased from the cell bank of Xiangya School of Medicine, Central South University; lung adenocarcinoma cell lines PC-9 and PC-9GR were purchased from Guangzhou Institute of Respiratory Diseases; breast cancer cell line MCF-7 was purchased from Emory University, USA Office of Professor You Shaojin, Faculty of Medicine.

[0124] 1.1.2 Main reagents and preparation

[0125] 1.1.2.1 Main experimental reagents and sources

[0126]

[0127]

[0128] 1.1.2.2 Preparation of main reagents

[0129] (1) Complete cell culture medium (sterile, stored in a 4-degree refrigerator for later use)

[0130] RPMI-1640 medium 90%

[0131] Fetal Bovine Serum 10%

[0132] Penicillin streptomycin double antibody 1%

[0133] (2) Cell cryopreservation solution (sterile, ready-to-use)

[0134] 9 parts of fetal bovine serum

[0135] 1 part of DMSO

[0136] (3) PBS solution (autoclaved, stored in a ...

Embodiment 1

[0247] 2.1 Expression of mPRα protein in different lung adenocarcinoma cells

[0248] In this study, three lung adenocarcinoma cell lines A549, PC-9 and PC-9GR with positive expression of mPRα were selected. We detected the expression of mPRα protein in these three cell lines by Western blot technique, the results are as follows: figure 1 Shown: breast cancer cell line MCF-7 is used as positive control, β-tubulin is used as internal reference, the expression of mPRα protein in A549 cells is positive, the expression of mPRα protein in PC-9 cells is weakly positive, and the expression of mPRα protein in PC-9GR cells is strongly positive. Compared with MCF-7 cells, the expression of mPRα protein in A549 cells, PC-9 cells, and PC-9GR cells was significantly different, *P<0.05, which further verified the three lung adenocarcinoma cell lines A549 selected in this experiment , PC-9 and PC-9GR were all mPRα positive cells. 2.2 Detection of EGFR mutation status in different lung ade...

Embodiment 2

[0260] 2.5 Inducing PC-9 cells to become drug-resistant PC-9GR cells with gefitinib

[0261] In order to dynamically observe the role of mPRα in the process of drug resistance of lung adenocarcinoma cells, we simulated the drug resistance environment of tumors in vivo, and artificially induced EGFR-TKIs-sensitive lung adenocarcinoma PC-9 cells in vitro with gradient increasing gefitinib Become drug-resistant strain PC-9GR cells. Before induction, we determined that the IC50 of gefitinib in PC-9 cells was 0.034uM, so we started to culture PC-9 cells at a low concentration (0.01uM), and changed the drug-containing medium every day. Change the drug-free medium for culture, and change to the drug-containing medium after the cells resume growth. If the cell proliferation is stable, consider entering the next concentration for induction, and the drug concentration of gefitinib used is 0.01uM-0.02uM-0.05uM-0.1uM-0.2uM-1.0uM-2.0uM-3.0uM. PC-9 cells can still proliferate stably until...

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Abstract

The invention discloses a method for adjusting the sensibility of lung adenocarcinoma cells to EGFR-TKIs by mediating progesterone by mPR alpha, according to the method disclosed by the invention, the effects on the sensibility of lung adenocarcinoma cells of different EGFR mutation states to the EGFR-TKIs caused by the mPR alpha are compared, by inducing lung adenocarcinoma sensitive strain PC-9 cells into drug-resistant strain PC-9GR cells, and the relationship of a drug-resistant process and mPR alpha expression is dynamically observed. According to he method for adjusting the sensibility of lung adenocarcinoma cells to the EGFR-TKIs by mediating the progesterone by the mPR alpha, whether the sensibility of lung adenocarcinoma cells of different EGFR mutation states to the EGFR-TKIs can be improved or not after the progesterone is mediated by a membrane receptor is discussed from the point of view of female sex hormone, and a theoretical basis is provided for future research of a lung adenocarcinoma target drug sensitizer.

Description

technical field [0001] The invention relates to the field of research on the relationship between sex hormones and receptors and lung adenocarcinoma, in particular to a method for regulating the sensitivity of lung adenocarcinoma cells to EGFR-TKIs by mediating progesterone with mPRα. Background technique [0002] In recent years, the role of female hormones and their receptors in the occurrence, development and treatment of lung adenocarcinoma has become a hot research topic. Some scholars believe that lung adenocarcinoma, like breast cancer and endometrial cancer, is also a sex hormone-dependent tumor. Female hormones include estrogen (Estrogen) and progesterone (progesterone, Progesterone, P4). The cancer-promoting effect of estrogen has reached a broad consensus, but the effect of progesterone on lung cancer seems to be different from that of estrogen. Ishibashi et al. found that treatment with progesterone could inhibit tumor growth in a progesterone receptor-positive ...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/68C12Q1/02G01N33/68
CPCC12N5/0693C12Q1/025C12Q1/6869C12Q1/6886C12Q2600/106C12Q2600/156G01N33/68
Inventor 陈琼卢晓晓李伟何淑雅肖锏
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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