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Bromocriptine drug-resistant cell strain for rat prolactinoma as well as preparation method and application of drug-resistant cell strain

A technology of prolactinoma and drug-resistant cells, which is applied in the field of preparation of bromocriptine-resistant cell lines of rat prolactinomas, can solve the problem of rare drug-resistant cell lines of prolactinomas and achieve high scientific research and production application value, the effect of stable drug resistance

Inactive Publication Date: 2018-11-13
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The rat prolactinoma MMQ cell line is a commonly used cell line for the study of prolactinomas, but the drug-resistant cell lines for prolactinomas are still rare, especially those induced by bromocriptine. No tumor drug-resistant cell lines have been reported

Method used

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  • Bromocriptine drug-resistant cell strain for rat prolactinoma as well as preparation method and application of drug-resistant cell strain
  • Bromocriptine drug-resistant cell strain for rat prolactinoma as well as preparation method and application of drug-resistant cell strain
  • Bromocriptine drug-resistant cell strain for rat prolactinoma as well as preparation method and application of drug-resistant cell strain

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Effect test

Embodiment 1

[0032] Example 1 Induction and Establishment of Rat Prolactinoma Bromocriptine-resistant Cell Line MMQ / BRC

[0033] The present invention uses rat prolactinoma MMQ cells as parent cells, adopts bromocriptine to simulate the chemotherapy process in the human body, and induces and establishes rat prolactinoma bromocriptine through repeated high-dose shocks combined with a method of gradually increasing drug concentration. Ting-resistant cell line MMQ / BRC. Specific steps are as follows:

[0034] (1) Rat prolactinoma MMQ cell line (purchased from Xiehe Cell Bank) was inoculated in DMEMF12 medium containing 10% fetal bovine serum, placed at 37°C, 5% CO 2 , cultured in an incubator with saturated humidity;

[0035] (2) Take MMQ cells in the logarithmic growth phase or when the cell density reaches 80%, inoculate them in fresh DMEM F12 medium containing 2 μmol / mL final concentration of bromocriptine, place at 37°C, 5% CO 2 , cultured in an incubator with saturated humidity, and ob...

Embodiment 2

[0049] The measurement of embodiment 2 MMQ and MMQ / BRC cell growth curve and drug resistance index

[0050] The growth curve and drug resistance index of the rat prolactinoma cell line and the bromocriptine-resistant cell line of the rat prolactinoma were measured respectively, and the specific steps were as follows:

[0051] 1. Take the rat prolactinoma cell line (MMQ) and rat prolactinoma bromocriptine-resistant cell line (MMQ / BRC) cells in good growth state, centrifuge at 1000rpm / min for 5min, and use serum-free Resuspend in DMEM / F12 medium and adjust the cell density to 1×10 5 / mL. The two kinds of cells were divided into blank control group (only adding culture medium), negative control group (cells were not added with drugs) and experimental group. They were seeded in 96-well plates at a volume of 90 μL per well, and starved for 12 hours without serum. experiment.

[0052] 2. Bromocriptine was dissolved in DMEM / F12 medium, the concentration of the concentrated solutio...

Embodiment 3

[0057] Example 3 MMQ and MMQ / BRC cell PRL secretion level

[0058] The expression levels of PRL in rat prolactinoma cell lines and rat prolactinoma bromocriptine-resistant cell lines were determined by ELISA, and the specific steps were as follows:

[0059] 1. Take MMQ and MMQ / BRC cells in the logarithmic growth phase, and use 1×10 6 / mL density inoculated in 6-well plate, 2mL per well. The cells were collected at 12, 24, 36, and 48 hours, centrifuged, and the supernatant was taken and stored at -80°C.

[0060] 2. Use the rat prolactin ELISA kit to detect the PRL in the collected supernatant. When detecting, first add 100 μL of samples and standards to the corresponding reaction wells, mix well and incubate at 37°C for 2 hours, and wash the plate 5 times with washing solution. .

[0061] 3. Add 100 μL of primary antibody, incubate at 37°C for 1 hour, and wash the plate 5 times.

[0062] 4. Add 100 μL enzyme-labeled secondary antibody to each well, incubate at 37°C for 30 m...

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Abstract

The invention belongs to the technical field of tumor biology and discloses a method for preparing a bromocriptine drug-resistant cell strain for rat prolactinoma. Rat prolactinoma MMQ cells are takenas parent cells, the drug administration process inside the human body is simulated through bromocriptine, and the bromocriptine drug-resistant cell strain MMQ / BRC for rat prolactinoma is establishedand acquired through induction with a method of repeated large-dose impact and gradual drug concentration increase. The bromocriptine drug-resistant cell strain for rat prolactinoma, which is prepared with the method, has the advantage of stable drug resistance, can be used for researching the characteristics of the morphology and biology of bromocriptine drug-resistant cells of human prolactinoma, researching the tumor drug resistant mechanism and analyzing the sensitivity of anti-tumor drugs and screening and evaluating anti-tumor drugs, can also be used for preparing a drug-resistant reversal agent or drug for tumor and has higher scientific research and production application value.

Description

technical field [0001] The invention relates to the technical field of tumor biology, namely a preparation method and application of a rat prolactinoma bromocriptine drug-resistant cell line. Background technique [0002] As a benign tumor, pituitary adenoma accounts for about 10-15% of all intracranial tumors. According to the hormone secretion activity in the body, it can be divided into functional type and non-functional type, and according to the tumor size, it can be divided into microadenomas, macroadenomas and giant adenomas. Prolactinoma, the most common type of pituitary adenoma, produces excessive prolactin secretion and a range of symptoms. According to autopsy and radiological studies, the incidence is about 40 to 57%. Although prolactinoma is a benign tumor, due to its hyperprolactinemia and intracranial compression, it can produce severe clinical symptoms: headache, visual disturbance, cranial nerve dysfunction, menstrual disturbance in women, amenorrhea, lac...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693C12N2501/06C12N2503/02G01N33/5011G01N2500/10
Inventor 朱永红简孟瑶
Owner SUN YAT SEN UNIV
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